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  • Immunocytochemistry  (120)
  • gene expression  (86)
  • Springer  (206)
  • 2020-2024
  • 1995-1999  (135)
  • 1985-1989  (63)
  • 1980-1984
  • 1975-1979  (8)
  • 1925-1929
  • 2022
  • 1995  (135)
  • 1988  (63)
  • 1976  (8)
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  • 2020-2024
  • 1995-1999  (135)
  • 1985-1989  (63)
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  • 1975-1979  (8)
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  • 1
    Electronic Resource
    Electronic Resource
    Heart anatomy and developmental biology (1988)
    Springer
    Cellular and molecular life sciences 44 (1988), S. 910-919 
    Details
    ISSN: 1420-9071
    Keywords: Heart embryology ; developmental biology ; differentiation ; morphogenesis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subject of heart development has attracted the interest of many embryologists over the last two centuries. As a result, the main morphologic features of the developmental anatomy of the heart are already well established. Although there are still some controversial points, and there is probably much descriptive work yet to be done, emphasis is currently being placed on developmental mechanisms rather than simply on descriptive facts. The availability of new techniques and the overall advances in biological research are placing heart embryology in a new perspective. Today, we do not simply ask whether one or another embryonic structure arises further right or further left; instead, we are studying how cells, tissues, and their microenvironment interrelate at the several levels of biological organization (from the gene upwards) so as to give rise to a mature organ with a distinct shape and well-established functions. This paper attempts to review some of the basic aspects of the developmental anatomy of the heart. Descriptive embryology is used here as a tool. Emphasis is placed on developmental mechanisms, and on the present knowledge of how these mechanisms are related to the structural development of the heart.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01939884
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  • 2
    Electronic Resource
    Electronic Resource
    A molecular view of cardiogenesis (1988)
    Springer
    Cellular and molecular life sciences 44 (1988), S. 930-936 
    Details
    ISSN: 1420-9071
    Keywords: Cardiac development ; embryogenesis ; gene expression ; complementary DNA ; molecular methodologies ; myocardial contractility ; myosin ; atrial natriuretic factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cardiac development involves a complex integration of subcellular processes into multicellular and, finally, whole organ effects. Until recently it has been difficult to investigate the genetic control of this organ level differentiation of the heart. The proliferation of molecular biology methodologies has provided mechanisms to directly investigate the control of these processes. This article focuses on molecular lines of research on two key areas in cardiac development: the regulation of expression of sarcomeric contractile and regulatory proteins, and atrial natriuretic factor. Molecular approaches are described which have allowed investigators to begin to determine the tissue and stage-specific expression of genes, to locate those genes in the genome, determine their sequences, and to directly investigate the mechanisms controlling their expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01939886
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  • 3
    Electronic Resource
    Electronic Resource
    Analysis of gene function inDictyostelium (1995)
    Springer
    Cellular and molecular life sciences 51 (1995), S. 1116-1123 
    Details
    ISSN: 1420-9071
    Keywords: Antisense RNA ; gene expression ; insertional mutagenesis ; physical mapping ; reporter genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Over the past ten years, powerful molecular genetic techniques have been developed to analyze gene function inDictyostelium. DNA-mediated transformation using a variety of selections and vectors has allowed the introduction of wild-type or modified genes that are under various forms of transcriptional control. Homologous recombination is efficient and can be used to modify the genome in precise ways. In addition, it is now possible to clone genes based on their mutant phenotype alone, either by insertional mutagenesis, or by screening antisense expression cDNA libraries. Finally, a nearly complete physical map of the genome is available and so genes are easily mapped by physical techniques. We discuss many of these advances within the context of major research problems presently under study.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01944729
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  • 4
    Electronic Resource
    Electronic Resource
    Regulation of metallothionein gene expression in Cd- or Zn-adapted RK-13 cells (1995)
    Springer
    Cellular and molecular life sciences 51 (1995), S. 606-611 
    Details
    ISSN: 1420-9071
    Keywords: Metallothionein ; isometallothioneins ; gene expression ; rabbit kidney cell-line ; cadmium adaptation ; zinc adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We explored the molecular genetics underlying the massive induction of isoMTs by Zn2+ or Cd2+ in metal tolerant rabbit kidney (RK-13) sub-line cells, using band shift assays and Southern blotting analysis. In sub-line cells accommodated to intermediate metal concentrations (100 μM Zn2+; 1–20 μM Cd2+) evidence suggested that the increase in the capacity for isoMT synthesis is brought about by an increased binding activity of the nuclear transcription factors MTF-1 and Sp1. Using quantitative band shift analysis with a mouse MRE-d oligonucleotide probe, the binding of both transcription factors was found to be enhanced two to three times over the binding activity measured in the unexposed parental RK-13 cells. Their increase in binding activity is probably the cause of the overexpression of MT genes and the development of metal tolerance in these cells. In cells tolerant to the highest concentrations of metal the analysis of Southern blot signals revealed MT gene amplification to be the most probable cause of the increased MT production. Thus, in cells of sub-lines growing in the presence of 350 μM Zn2+, two of the isoMT genes were coordinately triplicated and in cells tolerant to 150 μM Cd2+ one isoMT gene was amplified two-fold.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02128753
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  • 5
    Electronic Resource
    Electronic Resource
    Infection and disintegration of vascular tissues of non-suberized roots of spruce byHeterobasidion annosum and use of antibodies for characterizing infection (1995)
    Springer
    Mycopathologia 129 (1995), S. 91-101 
    Details
    ISSN: 1573-0832
    Keywords: ELISA ; Endodermis ; H. annosum ; Immunocytochemistry ; Root rot ; Vascular tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vascular disintegration mainly of medulla rays of spruce roots is of major significance in root rot disease of spruce caused byH. annosum. Using seedling roots as an experimental model, the possible routes and initial host reactions preceding invasion of vascular tissues was investigated. Transmission electron microscopy showed that penetration through the endodermis was an obvious route but not without host resistance. Using antibodies againstH. annosum hyphal materials, some labelling of vascular tissues remote from sites of fungal colonization suggest the release of fungal secretory products partly active in tissue disintegration. Similarly, intense labelling was also observed in severely colonized host tissues at late stages of infection. Strong labelling recorded at 3 d p.i. mainly on fungal hyphae and scant gold particles on invaded host tissues could imply that induction of host antifungal metabolites may have been a late event. A correlation was found between total antigenic material in root homogenates measured by ELISA, density of tissue labelling by immunocytochemistry and severity of disease symptoms. The importance of this in relation to diagnosis of biotic root rot diseases in the field is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01103468
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  • 6
    Electronic Resource
    Electronic Resource
    Aberrant protamine 1/protamine 2 ratios in sperm of infertile human males (1988)
    Springer
    Cellular and molecular life sciences 44 (1988), S. 52-55 
    Details
    ISSN: 1420-9071
    Keywords: Human sperm ; human protamine ; protamine 1 ; protamine 2 ; protamine 3 ; infertility ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Protamines were extracted from the sperm of fertile and infertile human males and the relative proportion of protamines 1, 2, and 3 were determined by scanning microdensitometry following electrophoresis of total protamine in polyacrylamide gels. The proportion of the three protamines was found to be similar in sperm obtained from different normal males. The distribution of protamines in sperm obtained from a select group of infertile males producing an elevated level of large sperm heads, in contrast, was different from that of the fertile males.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01960243
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  • 7
    Electronic Resource
    Electronic Resource
    Immunocytochemical localization of two key enzymes of the 2-hydroxyglutarate pathway of glutamate fermentation in Acidaminococcus fermentans (1988)
    Springer
    Archives of microbiology 150 (1988), S. 504-508 
    Details
    ISSN: 1432-072X
    Keywords: Acidaminococcus fermentans ; Glutamate fermentation ; Electron microscopy ; Immunocytochemistry ; Post-embedding labelling ; Antibody-gold complexes ; Protein A-gold complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have investigated the in situ location of glutaconyl-CoA decarboxylase and 2-htdroxyglutaryl-CoA dehydratase in Acidaminococcus fermentans using the antibody-gold and protein A-gold techniques carried out as a post-embedding immunoelectron microscopic procedure. Polyclonal antisera were raised in rabbits against homogeneous fractions of the enzymes. Anaerobically grown cells of A. fermentans of the late exponential growth phase were fixed with 0.2% glutaraldehyde and 0.3% formaldehyde (final concentrations) in the growth medium. Dehydration of the cells was achieved with methanol. The cells were embedded in the low temperature embedding resin Lowicryl K4M. The markers indicative for antigenic sites of the two enzymes unequivocally demonstrate that the sodium pump glutaconyl-CoA decarboxylase is located at the cell periphery being a membrane-bound enzyme as expected whereas 2-hydroxyglutaryl-CoA dehydratase is a soluble cytoplasmic enzyme.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422295
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  • 8
    Electronic Resource
    Electronic Resource
    Expression of mRNA for vasoactive intestinal peptide in normal human colon and during inflammation (1995)
    Springer
    Molecular and cellular biochemistry 142 (1995), S. 1-7 
    Details
    ISSN: 1573-4919
    Keywords: vasoactive intestinal peptide ; ulcerative colitis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The availability of colon provides a ready source of human neurons. Among the products of nerve cell bodies, vasoactive intestinal peptide is a neuropeptide that serves as a marker of non-adrenergic, non-cholinergic inhibitory nerves in colon. These nerves have been proposed to be involved in regulation of immune function, secretion, and smooth muscle function. In previous work, we identified decreased tissue levels of vasoactive intestinal peptide in a disorder of chronic colonic mucosal inflammation, ulcerative colitis. We hypothesized that diminished gene expression of vasoactive intestinal peptide could result in decreased tissue levels of this neuropeptide. Sigmoid colon was obtained at surgery from controls (n=6) and patients with ulcerative colitis (n=6). Vasoactive intestinal peptide mRNA was quantified by Northern blot hybridization and tissue levels of vasoactive intestinal peptide were determined by radioimmunoassay. Tissue vasoactive intestinal peptide was decreased only in the mucosalsubmucosal layer of ulcerative colitis (p=.02). There was a single 1.7 kbase vasoactive intestinal peptide transcript identified in both control colon and ulcerative colitis. Normalized vasoactive intestinal peptide mRNA levels were increased by 260% in ulcerative colitis compared to controls (p〈.01). These observations suggest that decreased vasoactive intestinal peptide gene expression or abnormal post-transcriptional processing are not primary defects in this disorder of chronic inflammation. The findings support the alternative hypothesis that axonal degeneration in ulcerative colitis could result in increased expression of neuronal vasoactive intestinal peptide mRNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00928907
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  • 9
    Electronic Resource
    Electronic Resource
    Expression of hepatic calcium-binding protein regucalcin mRNA is elevated by refeeding of fasted rats: Involvement of glucose, insulin and calcium as stimulating factors (1995)
    Springer
    Molecular and cellular biochemistry 142 (1995), S. 35-41 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; insulin ; calcium ; gene expression ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of refeeding on the expression of Ca2+-binding protein regucalcin mRNA in the liver of fasted rats was investigated. When rats were fasted overnight, the hepatic regucalcin mRNA level was reduced about 70% of that in feeding rats. Refeeding produced a remarkable elevation of hepatic regucalcin mRNA level (about 150–170% of fasted rats). Liver regucalcin concentration was appreciably increased by refeeding, although it was not altered by fasting. The oral administration of glucose (2 g/kg body weight) to fasted rats caused a significant increase in hepatic regucalcin mRNA level. Moreover, hepatic regucalcin mRNA level was clearly elevated by a single subcutaneous administration of insulin (10 and 100 U/kg) to fasted rats. The hormonal effect was not further enhanced by the simultaneous administration of calcium chloride (250 mg Ca/kg) to fasted rats, although calcium administration stimulated regucalcin mRNA expression in the liver. The present study suggests that the expression of hepatic regucalcin mRNA stimulated by refeeding is significantly involved in the action of insulin and/or calcium as stimulating factors.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00928911
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  • 10
    Electronic Resource
    Electronic Resource
    Specific species and tissue differences for the gene expression of calcium-binding protein regucalcin (1995)
    Springer
    Molecular and cellular biochemistry 143 (1995), S. 67-71 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; gene distribution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The existence and expression of gene encoding the Ca2+-binding protein regucalcin in various species and tissues were investigated with Southern and Northern hybridization analyses using regucalcin cDNA (0.9 kb of open reading frame). Genomic Southern hybridization analysis demonstrated that regucalcin gene was widely conserved among higher animals including human, monkey, rat, mouse, dog, bovine, rabbit and chicken. The gene was not found in yeast. The Northern blot analysis of poly (A)+RNAs extracted from the liver of various species showed that regucalcin mRNA was predominantly expressed in rat and mouse, although the expression was also seen in human, bovine and chicken. Furthermore, the enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG indicated that hepatic regucalcin concentration was most pronounced in rat as compared with that of guinea pig, mouse and chicken. These observations show that the gene expression of regucalcin and its protein synthesis is unique in the liver of rats, suggesting the existence of a specific mechanism in demonstrating regucalcin synthesis from gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00925928
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  • 11
    Electronic Resource
    Electronic Resource
    17β-Estradiol stimulates the expression of hepatic calcium-binding protein regucalcin mRNA in rats (1995)
    Springer
    Molecular and cellular biochemistry 143 (1995), S. 137-141 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; estrogen ; gene expression ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of nuclear receptor-related hormones on the expression of hepatic calcium-binding protein regucalcin mRNA in rats was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb of open-reading frame). A single subcutaneons administration of 17β-estradiol (0.5, 1.0 and 2.0 mg/kg body weight) in rats induced a remarkable increase of regucalcin mRNA in liver; the level was about 200% of control at 24 h after the administration of 2.0 mg/kg. The increase showed about 350% even at 6 h after the administration. Meanwhile, hepatic regucalcin mRNA level was not appreciably altered by a single subcutaneous administration of thyroxine (T4) (20, 40 and 80 mg/kg) or hydrocortisone (10 and 30 mg/kg) in rats. The present study demonstrates that the expression of hepatic regucalcin mRNA is stimulated by estrogen action in the liver nuclei of rats.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01816947
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  • 12
    Electronic Resource
    Electronic Resource
    Thyroid hormone inhibits fatty acid synthase gene transcription in chicken liver (1995)
    Springer
    Molecular and cellular biochemistry 144 (1995), S. 105-108 
    Details
    ISSN: 1573-4919
    Keywords: fatty acid synthase ; gene expression ; and thyroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of triiodothyronine (T3) on regulation of fatty acid synthase in chicken liver was investigated. In hypothyroid animals, enzyme activity was about one half of that in euthyroid animals. T3 treatment increased the enzyme activity in hypothyroid animals. There is little difference in both the mRNA concentration and the transcription rate between euthyroid and hypothyroid animals. T3 treatment markedly decreased both the mRNA concentration and the transcription rate in euthyroid and hypothyroid animals. These results suggested that T3 maintained the normal level of enzyme expression primarily by stimulating the post-transcriptional step, while the transcription of the gene was inhibited by hyperthyroidism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00944388
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  • 13
    Electronic Resource
    Electronic Resource
    Thiol modulation of TNFα and IL-1 induced MnSOD gene expression and activation of NF-κB (1995)
    Springer
    Molecular and cellular biochemistry 148 (1995), S. 45-57 
    Details
    ISSN: 1573-4919
    Keywords: manganese ; superoxide dismutase ; gene expression ; hyperoxide lung injury ; nuclear factor kappa B
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract TNFα and IL-1 each can activate NF-κB and induce gene expression of manganese superoxide dismutase (MnSOD), a mitochondrial matrix enzyme which can provide critical protection against hyperoxic lung injury. The regulation of MnSOD gene expression is not well understood. Since redox status can modulate NF-κB and potential κB site(s) exist in the MnSOD promoter, the effect of thiols (including NAC, DTT and 2-ME) on TNFα and IL-1 induced activation of NF-κB and MnSOD gene expression was investigated. Activation of NF-kB and increased MnSOD expression were potentiated by thiol reducing agents. In contrast, thiol oxidizing or alkylating agents inhibited both NF-κB activation and elevated MnSOD expression in response to TNFα or IL-1. Since protease inhibitors TPCK and TLCK can inhibit NF-κB activation, we also investigated the effect of these compounds on MnSOD expression and NF-κB activation. TPCK and TLCK each inhibited MnSOD gene expression and NF-κB activation. Since the MnSOD promoter also contains anAP-1 binding site, the effect of thiols and thiol modifying agents on AP-1 activation was investigated. Thiols had no consistent effect onAP-1 activation. Likewise, some of the thiol modifying compounds inhibited AP-1 activation by TNFα or IL-1, whereas others did not. Since diverse agents had similar effects on activation of NF-κB and MnSOD gene expression, we have demonstrated that activation of NF-κB and MnSOD gene expression are closely associated and that reduced sulfhydryl groups are required for cytokine mediation of both processes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00929502
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  • 14
    Electronic Resource
    Electronic Resource
    Post-castration rebound of an androgen regulated prostatic gene (1988)
    Springer
    Molecular and cellular biochemistry 84 (1988), S. 3-15 
    Details
    ISSN: 1573-4919
    Keywords: prostate ; androgens ; gene expression ; in situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary After castration, the rat dorsolateral prostate M-40 mRNA initially decreased then rebounded to precastrated levels. The cellular site of M-40 expression and its renewed expression after castration was defined by in situ hybridization histochemistry. In situ hybridization with either a 32P-labeled or biotin-labeled M-40 cDNA probe demonstrated that M-40 mRNA levels were higher in the lateral than dorsal prostate. A second androgen regulated gene, RWB, also was highly expressed in the lateral prostate. The biotinylated cDNA probes provided microscopic resolution of the expressing cells, revealing two distinct morphologies of lateral epithelium which expressed both the M-40 and RWB mRNA. These morphologies appeared in ducts which contained either epithelial cell sheets that were highly convoluted or thinner epithelial cells with a minimal degree of convolution. The RWB mRNA decreased in both cell populations in response to androgen withdrawal. The decline and reappearance of M-40 mRNA also appeared in both epithelial cell types. These data demonstrated that after castration the M-40 mRNA initially decreased as expected for an androgen sensitive gene and then progressed to a fully inducible state. The mechanism of this progression remains to be elucidated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00235188
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  • 15
    Electronic Resource
    Electronic Resource
    Expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats: The stimulation by calcium administration (1995)
    Springer
    Molecular and cellular biochemistry 146 (1995), S. 71-77 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb of open-reading frame). Regucalcin mRNA was expressed in the kidney cortex, and this expression was clearly increased by a single intraperitoneal administration of calcium chloride solution (5–15 mg Ca/100 g body weight) in rats; this increase was remarkable at 60–120 min after the administration. Thyroparathyroidectomy (TPTX) caused a slight decrease of regucalcin mRNA levels in the kidney cortex. However, the administration of calcium (10 mg/100 g) in TPTX rats produced a clear increase of regucalcin mRNA levels in the kidney cortex. The subcutaneous administration of calcitonin (10–100 MRC mU/100 g) or parathyroid hormone [1–34] (1–10 U/100 g) in TPTX rats which received calcium (10 mg/100 g) administration did not cause an appreciable alteration of regucalcin mRNA levels in the kidney cortex, suggesting that the mRNA expression is not stimulated by calcium-regulating hormones. The administration of trifluoperazine (TFP; 5 mg/100 g), an inhibitor of Ca2+/calmodulin action, completely blocked the expression of regucalcin mRNA stimulated by calcium administration. Now, calcium content in the kidney cortex was significantly elevated by a single intraperitpneal administration of calcium (10 mg/100 g) in rats. The present study clearly demonstrates that the expression of regucalcin mRNA in the kidney cortex is stimulated by calcium administration in rats. This expression may be mediated through Ca2+/calmodulin action in the kidney cortex.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00926884
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  • 16
    Electronic Resource
    Electronic Resource
    Suppressed expression of calcium-binding protein regucalcin mRNA in the renal cortex of rats with chemically induced kidney damage (1995)
    Springer
    Molecular and cellular biochemistry 151 (1995), S. 55-60 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium ; gene expression ; kidney damage ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The alteration of Ca2+-binding protein regucalcin mRNA expression in the kidney cortex of rats administered cisplatin and cephaloridine, which can induce kidney damage, was investigated. Cisplatin (0.25, 0.5 and 1.0 mg/100 g body weight) or cephaloridine (25, 50 and 100 mg/100 g) was intraperitoneally administered in rats, and 1, 2 and 3 days later they were sacrificed. The alteration in serum findings after the administration of cisplatin (1.0 mg/100 g) or cephaloridine (50 and 100 mg/100 g) demonstrated chemically induced kidney damage; blood urea nitrogen (BUN) concentration increased markedly and serum inorganic phosphorus or calcium concentration decreased significantly. Moreover, the administration of cisplatin (1.0 mg/100 g) or cephaloridine (100 mg/100 g) caused a remarkable increase of calcium content in the kidney cortex of rats, indicating kidney damage. The expression of regucalcin mRNA in the kidney cortex was markedly reduced by the administration of cisplatin or cephaloridine in rats, when the mRNA levels were analyzed by Northern blotting using rat liver regucalcin cDNA (0.9 kb). The mRNA decreases were seen with the used lowest dose of cisplatin or cephaloridine. The present study clearly demonstrates that the mRNA expression of Ca2+-binding protein regucalcin in the kidney cortex of rats is decreased by chemically induced kidney damage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01076896
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  • 17
    Electronic Resource
    Electronic Resource
    Molecular remodelling in hypertrophied hearts from polyomavirus large T-antigen transgenic mice (1995)
    Springer
    Molecular and cellular biochemistry 152 (1995), S. 131-141 
    Details
    ISSN: 1573-4919
    Keywords: gene expression ; mRNA ; proto-oncogenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Polyomavirus large T-antigen transgenic mice develop cardiac hypertrophy characterized by an increase in atrial natriuretic factor and β-myosin heavy chain isoform expression. The aim of this study was to examine changes in proto-oncogene expression in hypertrophied hearts from the transgenic mice. Expression of early growth response-1 (Egr-1) mRNA was detected in hearts from all 15 transgenic mice, but was not detectable in 13 control mice. Reverse transcriptase-polymerase chain reaction experiments usingEgr-1-specific primers confirmed the increase inEgr-1 mRNA in enlarged hearts from the transgenic mice. Expression of c-jun,junD and Ha-ras mRNAs was increased in the transgenic hearts 3, 17 and 2.8-fold, respectively. Western blots showed an increase in c-myc, c-jun and ras protein in hypertrophied transgenic hearts. Immunofluorescence analyses confirmed an increase in Egr-1 and c-jun protein in transgenic cardiomyocytes. Proliferating cell nuclear antigen, Ki-ras and HSP 90 mRNAs were decreased 22, 2.7 and 3-fold, respectively in the transgenic hearts. Not altered in most hypertrophied hearts was expression of c-fos, junB, p53, c-neu, c-myc, HSP70, HSP27, TGF-β or IGF-1 mRNAs. Proto-oncogene and growth factor gene expression in hypertrophy induced by PVLT expression is modulated, with some proto-oncogenes increased and others decreased in expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01076075
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  • 18
    Electronic Resource
    Electronic Resource
    The cat hemoglobin polymorphism: Southern blot analysis of theβ-globin gene region from cats of various Hb A/Hb B phenotypes (1988)
    Springer
    Biochemical genetics 26 (1988), S. 239-248 
    Details
    ISSN: 1573-4927
    Keywords: cat hemoglobins ; polymorphism ; DNA ; restriction endonuclease ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The molecular basis for the genetic control of variable proportions of the two cat hemoglobins Hb A(α2β 2 A ) and Hb B (α2β 2 B ) was investigated. Ratios of Hb A/Hb B vary between 50/50 and 90/10 among members of the mongrel cat population, with clusters around 50, 35, and 10% Hb B. Genomic DNA from cats of 50/50, 70/30, and 90/10 phenotypes were cut by restriction endonucleasesHindIII,EcoRI,BamHI,Bg1II,and Pst1 and hybridized to a fragment of the human β-globin gene. The results of the Southern blots suggested a pattern of homozygote, heterozygote, homozygote for the respective cat phenotypes, 50/50, 70/30, and 90/10. Therefore, the cat hemoglobin polymorphism seems to result from the possible combinations of an allelic gene pair.
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    URL: http://dx.doi.org/10.1007/BF00561463
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  • 19
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    Structural organization and differential expression of carrot β-fructofuranosidase genes: identification of a gene coding for a flower bud-specific isozyme (1995)
    Springer
    Plant molecular biology 28 (1995), S. 189-194 
    Details
    ISSN: 1573-5028
    Keywords: β-fructofuranosidase ; invertase ; gene expression ; gene structure ; flower buds ; Daucus carota
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three genomic clones (Inv *Dc1, Inv *Dc2 and Inv *Dc3) were isolated by using the cDNA for carrot cell wall β-fructofuranosidase as a probe. The expression patterns of the three genes differed markedly. High levels of Inv *Dc1 transcripts were found in leaves and roots of young carrot, whereas in plants with developing tap roots no transcripts were detected. A high level of mRNA of Inv *Dc1 was also present in suspension-cultured cells. In developing reproductive organs, only low levels of transcripts of Inv *Dc1 were found in flower buds and flowers and none at later stages of development. In contrast, Inv *Dc2 and Inv *Dc3 were not expressed in vegetative plant organs. Invb1 *Dc1 was exclusively and strongly expressed in flower buds, and Inv *Dc3 at a very low level in suspension-cultured cells.
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    URL: http://dx.doi.org/10.1007/BF00042049
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    High levels of ripening-specific reporter gene expression directed by tomato fruit polygalacturonase gene-flanking regions (1995)
    Springer
    Plant molecular biology 28 (1995), S. 423-435 
    Details
    ISSN: 1573-5028
    Keywords: fruit ; gene expression ; promoter ; ripening ; tomato ; transgenic plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 1.4 kb 5′ polygalacturonase (PG) gene-flanking region has previously been demonstrated to direct ripening-specific chloramphenicol acetyl transferase (CAT) expression in transgenic tomato plants. The steady state level of CAT mRNA in these plants was estimated to be less than 1% of the endogenous PG mRNA. Further constructs containing larger PG gene-flanking regions were generated and tested for their ability to direct higher levels of reporter gene expression. A 4.8 kb 5′-flanking region greatly increased levels of ripening-specific reporter gene activity, while a 1.8 kb 3′ region was only shown to have a positive regulatory role in the presence of the extended 5′ region. Transgenic plants containing the CAT gene flanked by both of these regions showed the same temporal pattern of accumulation of CAT and PG mRNA, and steady-state levels of the transgene mRNA were equivalent to 60% of the endogenous PG mRNA on a per gene basis. The proximal 150 bp of the PG promoter gave no detectable CAT activity. However, the distal 3.4 kb of the 4.8 kb 5′ PG promoter was shown to confer high levels of ripening-specific gene expression when placed in either orientation upstream of the 150 bp minimal promoter. The DNA sequence of the 3.4 kb region revealed a 400 bp imperfect reverse repeat, and sequences which showed similarity to functionally significant sequences from the ripening-related, ethylene-regulated tomato E8 and E4 gene promoters. The possible roles of the flanking regions in regulating PG gene expression are discussed.
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    URL: http://dx.doi.org/10.1007/BF00020391
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    Organization, inheritance and expression of acetohydroxyacid synthase genes in the cotton allotetraploid Gossypium hirsutum (1995)
    Springer
    Plant molecular biology 28 (1995), S. 837-846 
    Details
    ISSN: 1573-5028
    Keywords: acetohydroxyacid synthase ; gene organization ; gene expression ; herbicide resistance ; cotton ; Gossypium hirsutum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The acetohydroxyacid synthase (AHAS) gene family of the cotton AD allotetraploid Gossypium hirsutum has been cloned and characterized. We have identified six different AHAS genes from an analysis of genomic clones and Southern blots of genomic DNA. Four of the six genes are organized as tandem pairs, in which the genes are separated by only 2–3 kb. Conservation of restriction fragment length polymorphisms between G. hirsutum and A-genome and D-genome-containing diploid cottons was sufficient to assign the single genes in clones A5 and A19 to the A and D subgenomes, respectively. Each diploid genome has one tandem pair, but in these cases we could not make specific subgenomic assignments. DNA and deduced amino acid sequences were determined for the A5 and A19 genes, and an AHAS cDNA clone isolated from a leaflibrary. The sequence of the A19 gene matches that of the cDNA clone, while the A5 gene is 97.8% similar. The four genes comprising the tandem pairs are much less similar to the cDNA clone. The deduced amino acid sequences of the mature polypeptides encoded by the A5 and A19 genes are collinear with the housekeeping forms of AHAS from Arabidopsis thaliana, Nicotiana tabacum and Brassica napus. The constitutive expression of A5 and A19 was confirmed with RNase protection assays and northern blots. We conclude that these genes encode the main house-keeping froms of AHAS in G. hirsutum. Among the four AHAS genes comprising the two tandem pairs, at least two are functional. These genes exhibit either low-level constitutive expression (one or both of the ‘downstream’ genes of each pair), or highly specific expression in reproductive tissue (one or both of the ‘upstream’ genes of each pair). The AHAS gene family of G. hirsutum is more complex than that of other plants so far examined.
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    URL: http://dx.doi.org/10.1007/BF00042069
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    Isolation of cDNA clones of genes with altered expression levels in phosphate-starved Brassica nigra suspension cells (1995)
    Springer
    Plant molecular biology 28 (1995), S. 859-870 
    Details
    ISSN: 1573-5028
    Keywords: Brassica ; phosphate starvation ; gene expression ; β-glucosidase ; mineral nutrition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Differential gene expression at the transcriptional level was examined as an initial step in the investigation of the Pi starvation response of Brassica nigra suspension cells. Total RNA was extracted from 7-day old cells grown in media containing either no Pi, 1.25 mM or 10 mM Pi., In vitro translation was carried out using their respective poly(A)+ RNA isolates and the resultant polypeptides were separated on a high-resolution SDS-PAGE gel. Scanning densitometry identified four polypeptides (ca. 31.7, 32.3, 52.5 and 64.8 kDa) present only in the Pi-starved samples. Screening by differential hybridization was performed on a cDNA library constructed from mRNA isolated from Pi-starved cells. Probes prepared from mRNA from Pi-deficient and Pi-sufficient cells identified a number of clones representing mRNA species that were preferentially transcribed under Pi deficiency. These phosphate starvation-responsive (psr) clones were placed into eleven groups as determined by cross-hybridization. Northern blots showed that the corresponding genes are inducible in both mild and severe Pi starvation conditions. Preliminary sequencing identified one of the clones as being homologous to β-glucosidases from several plant species. The possible role of β-glucosidase during Pi starvation and the identities of the other psr genes are discussed.
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    URL: http://dx.doi.org/10.1007/BF00042071
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    Molecular cloning of two isoinhibitor forms of chymotrypsin inhibitor 1 (CI-1) from barley endosperm and their expression in normal and mutant barleys (1988)
    Springer
    Plant molecular biology 10 (1988), S. 521-535 
    Details
    ISSN: 1573-5028
    Keywords: barley ; chymotrypsin inhibitor ; gene expression ; endosperm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Full-length cDNA clones for barley chymotrypsin inhibitor 1 (CI-1) have been isolated from an endosperm-specific cDNA library. Hybridization and nucleotide sequence analyses indicate that these cDNAs represent two distinct types of CI-1 mRNA which we have called CI-1A and CI-1B. Both mRNAs encode polypeptides of 83 residues (M r=8790 and 8960) which differ at eleven positions. The full-length cDNA sequences do not predict N-terminal signal peptide extensions indicating that CI-1 is synthesized in the mature form in contrast to the homologous proteinase inhibitors of tomato and potato. Northern hybridization experiments show that the CI-1 genes are under strict developmental and organ-specific control. CI-1 transcripts were first detected in the developing barley endosperm between 12 and 14 days after anthesis but no CI-1-related sequences were detected in the RNA preparations from shoots, leaves or roots. The expression of CI-1 was also studied in the high-lysine barley mutants Hiproly, Risø 56 and Risø 1508. Approximately 15-fold (Hiproly) and 4-fold (Risø 56 and 1508) higher levels of CI-1 mRNA were detected in the mutant endosperms compared to normal barley. These results correlate well with the increased deposition of CI-1 in the high-lysine lines and show that the differential expression is controlled mainly at the level of transcription or stability of the mRNA. Using Southern-blots of barley DNA we estimate that there are three copies of CI-1 per haploid genome in both normal and mutant barley lines.
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    URL: http://dx.doi.org/10.1007/BF00033607
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    Isolation of an alfalfa histone H3 gene: structure and expression (1988)
    Springer
    Plant molecular biology 11 (1988), S. 641-649 
    Details
    ISSN: 1573-5028
    Keywords: codon usage ; gene expression ; histone H3 gene ; Medicago sativa ; somatic embryos
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A histone H3 gene was isolated from a dicotyledonous plant, alfalfa (Medicago sativa). The sequence analysis of this gene revealed no obvious GC preference in its codon usage. Apart from containing most of the typical consensus sequences found in both animal and plant histone genes, the alfalfa H3 gene exhibits distinct structural features such as (1) the unusual location of two GATCC motifs in its 5′ flanking sequence, (2) the existence of a CGCGGATC on the nonsense strand at position −232, (3) the existence of a long palindromic structure, and (4) several polyadenylation signal-like sequences in the 3′ flanking region. There are about 160 copies of histone H3 gene in alfalfa tetraploid genome. Using the alfalfa H3 gene as a probe to study the pattern of histone H3 transcripts in the alfalfa, we found that the H3 RNAs are undetectable in leaves, more in stems than in roots, and highest in somatic embryos. Moreover, the RNA products of H3 genes in all alfalfa tissues tested show unusually long nontranslated region compared to those of animal histone genes. An additional high molecular weight species of H3 transcript was detected only in somatic embryos.
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    URL: http://dx.doi.org/10.1007/BF00017464
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    Molecular cloning of two tandemly arranged peroxidase genes from Populus kitakamiensis and their differential regulation in the stem (1995)
    Springer
    Plant molecular biology 28 (1995), S. 677-689 
    Details
    ISSN: 1573-5028
    Keywords: linked gene ; gene expression ; peroxidase ; Populus kitakamiensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genomic library was prepared from Populus kitakamiensis and screened with the cDNA for an anionic peroxidase from P. kitakamiensis. One genomic clone was isolated that contained two tandemly oriented genes for anionic peroxidases, prxA3a and prxA4a. Both genes consisted of four exons and three introns; the introns had consensus nucleotides, namely, GT and AG, at their 5′ and 3′ ends, respectively. The prxA3a and prxA4a genes encoded 347 and 343 amino acid residues, respectively, including putative signal sequences at the amino-termini. Putative promoters and polyadenylation signals were found in the flanking regions of both genes. The sequence of the coding region of prxA3a was completely identical to that of the cDNA clone pA3, whereas the sequence of the coding region of prxA4a was only 73% identical to that of the cDNA clone pA3. Northern blot analysis showed that the patterns of expression of the mRNAs that corresponded to prxA3a and prxA4a differed in stems of P. kitakamiensis.
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    URL: http://dx.doi.org/10.1007/BF00021193
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    Gerbera hybrida (Asteraceae) imposes regulation at several anatomical levels during inflorescence development on the gene for dihydroflavonol-4-reductase (1995)
    Springer
    Plant molecular biology 28 (1995), S. 935-941 
    Details
    ISSN: 1573-5028
    Keywords: anthocyanin ; Compositae ; corolla ; dfr ; flower development ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the ornamental cut flower plant Gerbera hybrida the spatial distribution of regulatory molecules characteristic of differentiation of the composite inflorescence is visualized as the various patterns of anthocyanin pigmentation of different varieties. In order to identify genes that the plant can regulate according to these anatomical patterns, we have analysed gene expression affecting two enzymatic steps, chalcone synthase (CHS) and dihydroflavonol-4-reductase (DFR), in five gerbera varieties with spatially restricted anthocyanin pigmentation patterns. The dfr expression profiles vary at the levels of floral organ, flower type and region within corolla during inflorescence development according to the anthocyanin pigmentation of the cultivars. In contrast, chs expression, although regulated in a tissue-specific manner during inflorescence development, varies only occasionally. The variation in the dfr expression profiles between the varieties reveals spatially specific gene regulation that senses the differentiation events characteristic of the composite inflorescence.
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    URL: http://dx.doi.org/10.1007/BF00042077
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    Tissue-specific expression of a pea legumin gene in seeds of Nicotiana plumbaginifolia (1988)
    Springer
    Plant molecular biology 10 (1988), S. 203-214 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; gene expression ; legumin (Pisum) ; Nicotiana ; seed storage protein ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 3.4-kilobase genomic DNA fragment from Pisum sativum L. containing the LegA gene, which encodes a major legumin storage protein, was transferred to Nicotiana plumbaginifolia using an Agrobacterium tumefaciens strain containing the Bin 19 binary vector system. Northern hybridisation analysis of legA-transformed plants demonstrated that legumin-specific RNA was present in developing seeds but not in developing leaves. Legumin protein was immunologically detected in the mature seeds of legA-transformed plants, and was present as the correct-size protein composed of disulphide-bonded polypeptides. It is concluded that the transferred pea genomic fragment contains all the information necessary for seed-specific expression of the legA gene, and for correct processing of the primary transcript and the precursor legumin protein.
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    URL: http://dx.doi.org/10.1007/BF00027397
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    Cytoplasmic HSP70 homologues of pea: differential expression in vegetative and embryonic organs (1995)
    Springer
    Plant molecular biology 27 (1995), S. 441-456 
    Details
    ISSN: 1573-5028
    Keywords: HSP70 ; HSC70 ; seed development ; imbibition ; chaperone ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Eukaryotes express several cytoplasmic HSP70 genes, and their encoded proteins participate in diverse cellular processes. Three cDNAs encoding highly expressed cytoplasmic HSP70 homologues from Pisum sativum were cloned and characterized. They were designated PsHSP71.2, PsHSC71.0, and PsHSP70b. These HSP70 genes have different expression profiles in leaves: PsHSP71.2 is observed only in response to heat stress, PsHSC71.0 is present constitutively, and PsHSP70b is weakly constitutively expressed, but induced strongly in response to heat stress. In addition to being heat induced, the PsHSP71.2 mRNA is also expressed in zygotic, but not maternal organs of developing pea seeds, while PsHSC71.0 and PsHSP70b mRNAs are present in maternal and zygotic organs throughout seed development. Immunoblot analysis of parallel protein samples detects a 70 kDa polypeptide in all samples, and a 72 kDa polypeptide that corresponds to the PsHSP71.2 gene product is observed in cotyledons beginning at mid-maturation and in axes beginning between late maturation and desiccation. This polypeptide is not detected in the seed coat. The 72 kDa polypeptide remains abundant in both cotyledons and axes through germination, but declines substantially between 48 and 72 h after the onset of imbibition. Differential control of HSP70 expression during heat stress, seed maturation, and germination is consistent with the hypothesis that there are functional distinctions between cytoplasmic HSP70s.
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    URL: http://dx.doi.org/10.1007/BF00019312
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    The isocitrate lyase gene of cucumber: Isolation, characterisation and expression in cotyledons following seed germination (1995)
    Springer
    Plant molecular biology 27 (1995), S. 487-497 
    Details
    ISSN: 1573-5028
    Keywords: Cucumis sativus ; gene expression ; glyoxylate cycle ; glyoxysome ; isocitrate lyase ; seed germination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cucumber (Cucumis sativus L.) genome contains only a single gene encoding the glyoxylate cycle enzyme isocitrate lyase (ICL). The cucumber icl gene has been isolated and sequenced, revealing only two small introns. The predicted amino acid sequence is more than 85% identical to ICL from other higher plants, and contains the C-terminal tripeptide Ser-Arg-Met which resembles a peroxisomal targeting sequence. The icl gene is coordinately expressed with the malate synthase (ms) gene after seed germination in both the light and the dark, suggesting that these genes may contain similar DNA elements regulating transcription. The start of transcription of the icl gene was determined and the DNA sequences upstream compared with the region of the ms gene promoter known to regulate transcription. This comparison revealed a highly conserved DNA sequence at similar positions in each gene.
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    URL: http://dx.doi.org/10.1007/BF00019316
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    Isolation of a full-length mitotic cyclin cDNA clone CycIIIMs from Medicago sativa: Chromosomal mapping and expression (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1059-1070 
    Details
    ISSN: 1573-5028
    Keywords: Alfalfa ; cell division cycle ; chromosomal location ; cyclin ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyclins in association with the protein kinase p34cdc2and related cyclin-dependent protein kinases (cdks) are key regulatory elements in controlling the cell division cycle. Here, we describe the identification and characterization of a full-length cDNA clone of alfalfa mitotic cyclin, termed CycIIIMs. Computer analysis of known plant cyclin gene sequences revealed that this cyclin belongs to the same structural group as the other known partial alfalfa cyclin sequences. Genetic segregation analysis based on DNA-DNA hybridization data showed that the CycIIIMs gene(s) locates in a single chromosomal region on linkage group 5 of the alfalfa genetic map between RFLP markers UO89A and CG13. The assignment of this cyclin to the mitotic cyclin class was based on its cDNA-derived sequence and its differential expression during G2/M cell cycle phase transition of a partially synchronized alfalfa cell culture. Sequence analysis indicated common motifs with both the A- and B-types of mitotic cyclins similarly to the newly described B3-type of animal cyclins.
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    URL: http://dx.doi.org/10.1007/BF00020880
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    Molecular cloning and expression analysis of peroxidase genes from wheat (1995)
    Springer
    Plant molecular biology 29 (1995), S. 647-662 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; peroxidase ; powdery mildew ; splicing ; Triticum aestivum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A PCR-based screening approach was used to isolate genomic clones from wheat encoding peroxidase isozymes. Three complete genes (pox1, pox2 and pox4) and one truncated gene (pox3) were characterized. The nucleotide sequences predicted mature proteins of 31 kDa, in which all the highly conserved motifs of secreted plant peroxidases were preserved. The coding regions showed 73–83% DNA sequence identity, with the highest level of similarity noted for the tandemly oriented pox2 and pox3. Expression of respective pox genes in various tissues of wheat was assessed by the RT-PCR technique, which showed that all four genes are active. The primary pox1 mRNA was spliced to remove three introns, whereas processing of the other pox transcripts involved only two intervening sequences. Splicing occurred at consensus GU/AG splice sites except for the first introns of pox1, pox2 and pox4 transcripts, where processing took place at unusual GC donor sites. The RNA analysis suggested that the pox1, pox2 and pox4 genes are predominantly expressed in roots. Lower levels of expression were found for pox4 and pox3 in leaves. Infection of wheat by the powdery mildew fungus selectively induced expression of pox2 in leaves.
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    URL: http://dx.doi.org/10.1007/BF00041156
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    A non-photosynthetic ferredoxin gene is induced by ethylene in Citrus organs (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1211-1221 
    Details
    ISSN: 1573-5028
    Keywords: ferredoxin ; Citrus ; ethylene ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence and expression of mRNA homologous to a cDNA encoding a non-photosynthetic ferredoxin (Fd1) from Citrus fruit was investigated. The non-photosynthetic nature of this ferredoxin was deduced from: (1) amino acid sequence alignments showing better scores with non-photosynthetic than with photosynthetic ferredoxins, (2) higher expression in tissues containing plastids other than chloroplast such as petals, young fruits, roots and peel of fully coloured fruits, and (3) the absence of light-dark regulation characteristic of photosynthetic ferredoxins. In a phylogenetic tree constructed with higher-plant ferredoxins, Citrus fruit ferredoxin clustered together with root ferredoxins and separated from the photosynthetic ferredoxins. Non photosynthetic (root and fruit) ferredoxins, but not the photosynthetic ferredoxins, have their closest homologs in cyanobacteria. Analysis of ferredoxin genomic organization suggested that non-photosynthetic ferredoxins exist in Citrus as a small gene family. Expression of Fd1 is developmentally regulated during flower opening and fruit maturation, both processes may be mediated by ethylene in Citrus. Exogenous ethylene application also induced the expression of Fd1 both in flavedo and leaves. The induction of non-photosynthetic ferredoxins could be related with the demand for reducing power in non-green, but biosynthetically active, tissues.
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    URL: http://dx.doi.org/10.1007/BF00020463
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    The effect of matrix attached regions (MAR) and specialized chromatin structure (SCS) on the expression of gene constructs in cultured cells and in transgenic mice (1995)
    Springer
    Molecular biology reports 22 (1995), S. 37-46 
    Details
    ISSN: 1573-4978
    Keywords: MAR ; SCS ; insulation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The flanking sequences of several genes have been shown to direct a position independent expression of transgenes. Attempts to completely identify the insulating sequences have failed so far. Some of these sequences contain a matrix attached region (MAR) located in the flanking part of the genes. This article will show that the MARs in cultured cells located in the 3' OH region of the human apolipoprotein B100 (Apo B100) and within the SV40 genome were unable to stimulate and insulate transgene expression directed by the promoters from a rabbit whey acidic protein (WAP) gene or from human cytomegalovirus (hCMV) early genes. In transgenic mice, the MAR from the Apo B100 and SV40 genes did not enhance the expression of a transgene containing the rabbit whey acid protein (WAP) promotor, the late gene SV40 intron (VP1 intron), the bovine growth hormone (bGH) cDNA and the SV40 late gene terminator. This construct was even toxic for embryos. Similarly, the specialized chromatin structure (SCS) from the Drosophila 87A7 HSP70 gene reduced chloramphenicol acetyl transferase (CAT) activity when added between a cytomegalovirus (CMV) enhancer and a Herpes simplex thymidine kinase (TK) gene promoter. This inhibitory action was almost complete when a second SCS sequence was added before the CMV enhancer. Sequences from the firefly luciferase and from the human gene cathepsin D cDNA used as control unexpectedly showed a similar inhibitory effect when added to the CMVTKCAT construct instead of SCS. When added before the CMV enhancer and after the transcription terminator in the CMVTKCAT construct, the SCS sequence was unable to insulate the integrated gene as seen by the fact that the level of CAT in cell extracts were by no means correlated with the number of copies in individual clones. From these data, it is concluded that i) a MAR containing the canonical AT rich sequences does not amplify the expression of all gene constructs ii) AT rich MAR sequences do not have per se an insulating effect iii) Drosophila SCS from the 87A7 HSP70 gene has no insulating effect in all gene constructs (at least in mammalian cells) iv) and the addition of a DNA fragment between an enhancer and a promoter in a gene construct cannot be used as a reliable test to evaluate its insulating property.
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    URL: http://dx.doi.org/10.1007/BF00996303
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    Molecular cloning and analysis of four potato tuber mRNAs (1988)
    Springer
    Plant molecular biology 11 (1988), S. 255-269 
    Details
    ISSN: 1573-5028
    Keywords: potato ; patatin ; proteinase inhibitor ; cloning ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tuberization in potato is a complex developmental process involving the expression of a specific set of genes leading to the synthesis of tuber proteins. We here report the cloning and analysis of mRNAs encoding tuber proteins. From a potato tuber cDNA library four different recombinants were isolated which hybridized predominantly with tuber mRNAs. Northern blot hybridization experiments showed that three of them, pPATB2, p303 and p340, can be regarded as tuber-specific while the fourth, p322, hybridizes to tuber and stem mRNA. Hybrid-selected in vitro translation and nucleotide sequence analysis indicate that pPATB2 and p303 represent patatin and the proteinase inhibitor II mRNA respectively. Recombinant p322 represents an mRNA encoding a polypeptide having homology with the soybean Bowman-Birk proteinase inhibitor while p340 represents an mRNA encoding a polypeptide showing homology with the winged bean Kunitz trypsin inhibitor. In total, these four polypeptides constitute approximately 50% of the soluble tuber protein. Using Southern blot analysis of potato DNA we estimate that these mRNAs are encoded by small multigene families.
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    URL: http://dx.doi.org/10.1007/BF00027383
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    A homologue of the MAP/ERK family of protein kinase genes is expressed in vegetative and in female reproductive organs of Petunia hybrida (1995)
    Springer
    Plant molecular biology 27 (1995), S. 339-350 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; embryo sac ; ovule ; Petunia hybrida ; protein ; kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mitogen activated protein (MAP) kinase pathway of eukaryotes is stimulated by many growth factors and is required for the integration of multiple cellular signals. In order to study the function of MAP kinases during plant ovule development we have synthesized a Petunia hybrida ovule-specific cDNA library and screened for MAP protein kinase-related sequences using a DNA probe obtained by PCR. A full-length cDNA clone was identified (PMEK for Petunia hybrida MAP/ERK-related protein kinase) and shown to encode a protein related to the family of MAP/ERK protein kinases. Southern blot analysis showed that PMEK is a member of a small multigene family in P. hybrida. The cDNA codes for a protein (PMEK1) of 44.4 kDa with an overall sequence identity of 44% to the products of the mammalian ERK/MAP kinase gene, and the budding yeast KSS1 and FUS3 genes. PMEK1 displays 96 and 80% identity respectively with the tobacco NTF3 and Arabidopsis ATMPK1 kinases, and only 50% to the more distantly related plant MAP kinase MsERK1 from alfalfa. The two phosphorylation sites found in the loop between subdomain VII and VIII in all the other MAP kinases are also present in PMEK1. RNA gel blot and RT-PCR analyses demonstrated that PMEK1 is expressed in vegetative organs and preferentially accumulated in female reproductive organs of P. hybrida. In situ hybridization experiments showed that in the reproductive organs PMEK1 is expressed only in the ovary and not in the stamen.
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    URL: http://dx.doi.org/10.1007/BF00020188
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    Isolation and expression analysis of cDNA clones encoding a small and a large subunit of ADP-glucose pyrophosphorylase from sugar beet (1995)
    Springer
    Plant molecular biology 27 (1995), S. 191-197 
    Details
    ISSN: 1573-5028
    Keywords: ADP-glucose pyrophosphorylase (small and large subunit) ; DNA sequence ; gene expression ; starch synthesis ; sugar beet (Beta vulgaris L.)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cDNA cloning of a small and a large subunit of ADP-glucose pyrophosphorylase (AGPase) from sugar beet is reported. The deduced amino acid sequences are highly homologous to previously identified AGPase polypeptides from other plant species. Both subunits are encoded by low copy genes. When RNA gel blot experiments were performed, strongest expression was detected in sink and source leaves of greenhouse-grown sugar beet plants. A lower expression was found in other tissues tested, i.e. in the hypocotyl, the tap root and roots. In these tissues, slightly higher transcript levels were found for the small subunit gene than for the large subunit gene.
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    Expression of the betaine aldehyde dehydrogenase gene in barley in response to osmotic stress and abscisic acid (1995)
    Springer
    Plant molecular biology 27 (1995), S. 307-315 
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    ISSN: 1573-5028
    Keywords: glycine betaine ; betaine aldehyde dehydrogenase ; osmotic stress ; gene expression ; plant hormone ; abscisic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When subjected to salt stress or drought, some vascular plants such as barley respond with an increased accumulation of the osmoprotectant glycine betaine (betaine), being the last step of betaine synthesis catalyzed by betaine aldehyde dehydrogenase (BADH). We report here cloning and characterization of BADH cDNA from barley, a monocot, and the expression pattern of a BADH transcript. An open reading frame of 1515 bp encoded a protein which showed high homology to BADH enzymes present in other plants (spinach and sugar-beet) and in Escherichia coli. Transgenic tobacco plants harboring the clone expressed high levels of both BADH protein and its enzymatic activity. Northern blot analyses indicated that BADH mRNA levels increased almost 8-fold and 2-fold, respectively, in leaves and roots of barley plants grown in high-salt conditions, and that these levels decreased upon release of the stress, whereas they did not decrease under continuous salt stress. BADH transcripts also accumulate in response to water stress or drought, indicating a common response of the plant to osmotic changes that affect its water status. The addition of abscisic acid (ABA) to plants during growth also increased the levels of BADH transcripts dramatically, although the response was delayed when compared to that found for salt-stressed plants. Removal of plant roots before transferring the plants to high-salt conditions reduced only slightly the accumulation of BADH transcripts in the leaves.
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    URL: http://dx.doi.org/10.1007/BF00020185
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    Loss of chloroplast transcripts for proteins associated with photosystem II: an early event during heat-bleaching in Euglena gracilis (1995)
    Springer
    Plant molecular biology 27 (1995), S. 317-325 
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    ISSN: 1573-5028
    Keywords: chloroplasts ; gene expression ; heat bleaching ; photosynthesis ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A shift in the ratio of chlorophyll (Chl) a and Chl b is an early indicator of heat bleaching in Euglena gracilis. This observation prompted us to consider whether or not changes in steady-state levels of chloroplast transcripts and in transcriptional activity could limit the synthesis of Chl a-binding proteins in bleaching plastids. We found that the mature transcripts for CP47 and CP43, the Chl a-binding apoproteins of the proximal antenna of photosystem II, decline sharply very early during bleaching. Our study also shows that transcription of psbB and psbC, the chloroplast genes encoding CP47 and CP43, remains essentially unchanged during the same interval. We conclude that posttranscriptional events, such as mRNA stability, could play a major role in initiating an irreversible loss of chloroplast function in Euglena at a moderately elevated temperature. Lack of these transcripts would eventually impair the assembly of photosystem II in thylakoids.
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    Differential expression of two barley SNF1-related protein kinase genes (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1235-1240 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; gene family ; higher plants ; Hordeum vulgare ; metabolic regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have amplified and cloned DNA sequences derived from a gene encoding a SNF1 (sucrose-non-fermenting 1)-related protein kinase which differs from that previously reported from barley. Northern blot and polymerase chain reaction (PCR) analysis of RNA populations, using specific probes and oligonucleotide primers, indicated that the two SNF1-related genes are differentially regulated. One is expressed in all tissues, whereas the other is expressed at high levels in the seed endosperm and aleurone, but at levels undetectable by northern blot analysis in other tissues. Comparisons with other plant SNF1-related protein kinase genes suggest that the form which is expressed at greatly enhanced levels in the seed is less similar to the other plant homologues which have been reported and may be unique to cereals.
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    URL: http://dx.doi.org/10.1007/BF00020898
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    Evidence for some common signal transduction events for opposite regulation of nitrate reductase and phytochrome-I gene expression by light (1995)
    Springer
    Plant molecular biology 29 (1995), S. 25-35 
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    ISSN: 1573-5028
    Keywords: gene expression ; light regulation ; nitrate reductase ; phytochrome ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have explored the possible involvement of the phosphoinositide (PI) cycle and protein kinase C (PKC) in the phytochrome (Pfr)-mediated light signal transduction pathway using nitrate reductase (NR) and phytochrome-I (PhyI) genes as model systems. We have shown earlier that phorbol myristate acetate (PMA) completely replaces the red light effect in stimulating nitrate reductase activity and transcript levels in maize. In this paper, we present detailed evidence to show that PMA mimics the red light effect and follows similar kinetics to enhance NR steady-state transcript accumulation in a nitrate-dependent manner. We also show that PMA inhibits phyI steady-state transcript accumulation in a manner similar to red light, indicating that a PKC-type enzyme(s) may be involved in mediating the light effect in both cases. Serotonin or 5-hydroxytryptamine (5-HT), a stimulator of PI turnover, was also found to mimic the red light effect in enhancing NR transcript levels and inhibiting phyI transcript accumulation, indicating the role of the PI cycle in generating second messengers for regulating the two genes. These results indicate that phytochrome-mediated light regulation of NR and phyI gene expression may involve certain common steps in the signal transduction pathway such as the PI cycle and protein phosphorylation by a PKC-type enzyme.
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    URL: http://dx.doi.org/10.1007/BF00019116
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    Promoter analysis of the auxin-regulated tobacco glutathione S-transferase genes Nt103-1 and Nt103-35 (1995)
    Springer
    Plant molecular biology 29 (1995), S. 413-429 
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    ISSN: 1573-5028
    Keywords: auxin ; DNA binding factor ; gene expression ; glutathione S-transferase ; Nicotiana tabacum ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have analysed the promoter regions of two closely related auxin-regulated glutathione S-transferase genes. All active deletion constructs tested showed expression of the reporter gene β-glucuronidase (gusA) in root tips of young seedlings and newly developing lateral roots. Auxin treatment greatly enhanced the level of expression. The Nt103-1 promoter region −370/−276 was found to be necessary, at least as a quantitative element to confer auxin-responsiveness to a reporter gene, and sequences responsible for the auxin-responsiveness must be located downstream of −370. The region −651/−370 contains sequence information necessary for uninduced expression. The Nt103-35 promoter manifested its auxin-responsiveness within the −504/−310 region. Electrophoretic mobility shift analysis, using nuclear extracts from tobacco leaves and suspension cells, identified a factor binding to a sequence (ap103, TGAGTCT) at position −560 of the Nt103-1 promoter, which shows homology to the mammalian AP-1 site. A second factor was found to bind a sequence (as103, ATAGCTAAGTGCTTACG) with homology to the CaMV 35S promoter as-1 element. The as103 element is present in both promoters and positioned around −360, so within the region determined to be indispensable for the response to auxin. A third factor was found binding to the −276/−190 region of both promoters. Combined, these data point to the relevance of a 90 bp region for auxin-induced activity of both tobacco genes. The ASF-1 like factor binding to the as103 element within this region might be involved in mediating the auxin response.
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    A role for glutamate decarboxylase during tomato ripening: the characterisation of a cDNA encoding a putative glutamate decarboxylase with a calmodulin-binding site (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1143-1151 
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    ISSN: 1573-5028
    Keywords: differential screening ; gene expression ; Lycopersicon esculentum ; rin ; ripening inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A tomato fruit cDNA library was differentially screened to identify mRNAs present at higher levels in fruit of the tomato ripening mutant rin (ripening inhibitor). Complete sequencing of a unique clone ERT D1 revealed an open reading frame with homology to several glutamate decarboxylases. The deduced polypeptide sequence has 80% overall amino acid sequence similarity to a Petunia hybrida glutamate decarboxylase (petGAD) which carries a calmodulin-binding site at its carboxyl terminus and ERT D1 appears to have a similar domain. ERT D1 mRNA levels peaked at the first visible sign of fruit colour change during normal tomato ripening and then declined, whereas in fruit of the ripening impaired mutant, rin, accumulation of this mRNA continued until at least 14 days after the onset of ripening. This mRNA was present at much lower levels in other tissues, such as leaves, roots and stem, and was not increased by wounding. Possible roles for GAD, and its product γ-aminobutyric acid (GABA) in fruit, are discussed.
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    Isolation and characterisation of a melon cDNA clone encoding phytoene synthase (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1153-1162 
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    ISSN: 1573-5028
    Keywords: carotenoids ; cleavage site ; gene expression ; melon ; phytoene synthase ; ripening
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone (MEL5), encoding a protein homologous to phytoene synthase (PSY), has been isolated from a climacteric melon fruit cDNA library, using the tomato cDNA clone TOM5 [34] as a heterologous probe. MEL5 hybridised to a transcript of 1.65 kb which suggested that the 1.36 kb clone, isolated originally, was not full-length. The missing 5′ end was isolated by a reverse transcriptase-polymerase chain reaction (RT-PCR)-based method. This enabled the full sequence of the protein to be deduced and the cleavage site of the transit peptide for chromoplast import to be predicted. Northern analysis of RNA extracted from fruit samples of different ripening stages as well as from roots, leaves and flower petals was used to examine the expression pattern of the corresponding mRNA. The transcript corresponding to MEL5 is present at low quantities in unripe (green) fruit, reaches its highest levels when the fruit turns from green to orange and persists at lower levels during later ripening stages. A similar transcript was also detected in flower petals and in trace amounts in leaves and roots. Genomic Southern analysis indicates that the clone is homologous to a low-copy-number gene family. Sequence analysis showed a high degree of conservation among plant PSYs.
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    An unusual Group 2 LEA gene family in citrus responsive to low temperature (1995)
    Springer
    Plant molecular biology 29 (1995), S. 11-23 
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    ISSN: 1573-5028
    Keywords: drought ; flooding ; freezing tolerance ; gene expression ; salt
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Six cDNAs representing unique cold-induced sequences have been cloned from the hardy citrus relative Poncirus trifoliata. Among these, pBCORc115 and pBCORc119 were found to belong to the same gene family. Sequencing data indicated that pBCORc115 and pBCORc119 each contained an open reading frame, coding for a 19.8 kDa protein (COR19) and a smaller 11.4 kDa protein (COR11) respectively. Inspection of the deduced amino acid sequences revealed three large repeats in COR19, but only one was present in the COR11. Two elements: a Q-clustered tract and a K-rich motif were identified in each repeat. The K-rich motifs were similar to those of cotton D-11 and Group 2 LEA proteins. A Serine-cluster, a common feature in many Group 2 LEA-like proteins, was also found in these proteins, but it was in an unusual position at the carboxy-terminus. A bipartite motif of basic residues, similar to known nuclear targeting sequences, was also present in COR19 and COR11, suggesting that members of this protein family may have a nuclear targeting function. The expression of COR19 mRNA in response to cold acclimation, drought, flooding, and salinization was examined. COR19 expression in leaf tissue was induced in response to cold acclimation, but repressed during drought and flooding stress.
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    Abundance and half-life of the distinct oat phytochrome A3 and A4 mRNAs (1995)
    Springer
    Plant molecular biology 29 (1995), S. 367-377 
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    ISSN: 1573-5028
    Keywords: Avena sativa ; gene expression ; PHYA ; light regulation ; mRNA degradation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gene-preferential oligonucleotide probes were used to determined the relative abundance and half-lives of distinct oat phytochrome A (PHYA) mRNAs. Oat PHYA mRNAs are highly conserved in the 5′-untranslated region and the coding region, but the 3′-untranslated region has an overall lower sequence conservation and was the source of gene-preferential probes. PHYA3 mRNA was estimated to be ca. 61% of the oat PHYA mRNA pool present in poly(A)+ RNA from dark-grown seedlings. The half-lives for PHYA3 and PHYA4 mRNAs were both estimated to be ca. 30 min, and a similar short half-life was estimated for the average PHYA mRNA. Sequence comparisons of PHYA mRNAs from four grass species identified conserved sequences within the 5′- and 3′-untranslated regions that might be important for PHYA mRNA degradation.
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    Expression of arabidopsis cytosolic ascorbate peroxidase gene in response to ozone or sulfur dioxide (1995)
    Springer
    Plant molecular biology 29 (1995), S. 479-489 
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    ISSN: 1573-5028
    Keywords: Ascorbate peroxidase ; Arabidopsis thaliana ; gene expression ; guaiacol peroxidase ; ozone ; sulfur dioxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of ozone or sulfur dioxide on antioxidant enzymes were investigated in Arabidopsis thaliana. Plants were fumigated with 0.1–0.15 ppm ozone or sulfur dioxide up to about 1 week in an environment-controlled chamber. Both pollutants increased the activities of ascorbate peroxidase and guaiacol per-oxidase in leaves, but had little effect on the activities of superoxide dismutase, catalase, monodehydroascorbate reductase, dehydroascorbate reductase or glutathione reductase. Ozone was more effective than sulfur dioxide in increasing the activities of the peroxidases. Ascorbate peroxidase activity increased 1.8-fold without a lag period during fumigation with 0.1 ppm ozone, while guaiacol peroxidase activity increased 4.4-fold with a 1-day lag. Expression of the APX1 gene encoding cytosolic ascorbate peroxidase was further investigated. Its protein levels in leaves exposed to 0.1 ppm ozone for 4 or 8 days were 1.5-fold higher than in controls. Both ozone and sulfur dioxide elevated APX1 mRNA levels in leaves at 4 and 7 days, whereas at 1 day only ozone was effective. The induction of APX1 mRNA levels by ozone (3.4- to 4.1-fold) was more prominent than that by sulfur dioxide (1.6-to 2.6-fold). The APX1 mRNA level increased by day and decreased by night. Exposure of plants to 0.1 ppm ozone enhanced the APX1 mRNA level within 3 h, which showed a diurnal rhythm similar to that of the control. These results demonstrate that near-ambient concentrations of ozone as well as similar concentrations of sulfur dioxide can induce APX1 gene expression in A. thaliana.
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    Cloning and characterization of differentially expressed genes in imbibed dormant and afterripened Avena fatua embryos (1995)
    Springer
    Plant molecular biology 29 (1995), S. 823-831 
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    ISSN: 1573-5028
    Keywords: afterripening ; aldose reductase ; Avena fatua ; gene expression ; LEA ; seed dormancy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To analyze the patterns of gene expression associated with seed dormancy in wild oat (Avena fatua), we have isolated cDNA clones corresponding to genes that are differentially expressed in dormant and afterripened line M73 embryos. Gene transcripts of these clones were maintained in embryos of imbibed dormant caryopses, but declined rapidly in afterripened embryos after imbibition. GA3 treatment of dormant caryopses, which breaks dormancy, could lower the transcript levels in dormant embryos. When the germination of afterripened caryopses was inhibited by high temperature (35 °C), the decline in abundance of the transcripts in afterripened embryos was arrested. These genes were expressed to various degrees in water-stressed, but not in unstressed, 7-day-old seedlings. The expression of the genes was also ABA-inducible in afterripened embryos. The expression patterns in non-dormant line SH430 wild oat were similar to those of afterripened M73. DNA sequence analyses indicated that some of the cDNA clones encode LEA (late embryogenesis-abundant) proteins and aldose reductase. The significance of the expression of these genes in maintaining seed dormancy or longevity is discussed.
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    Molecular cloning and characterization of six cDNAs expressed during glucose starvation in excised maize (Zea mays L.) root tips (1995)
    Springer
    Plant molecular biology 28 (1995), S. 473-485 
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    ISSN: 1573-5028
    Keywords: carbon catabolite repression ; cDNA ; gene expression ; stress-induced genes ; glucose-starvation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to isolate glucose-starvation-related cDNAs in maize (Zea mays L.) root tips, a cDNA library was constructed with poly(A)+ mRNA from 24 h starved root tips. After differential screening of the library, we isolated six different cDNAs (named pZSS2 and pZSS7) which were expressed during glucose starvation. Time course analysis revealed that maximum expression of five of these genes occurs 30 h after the onset of the starvation treatment. On the contrary, the expression of mRNAs corresponding to pZSS4 was maximal at an early stage of starvation and then dramatically decreased. The expression of this gene did not seem to be specific for glucose starvation. The pattern of induction of the genes corresponding to pZSS2, pZSS3, pZSS5, pZSS6 and pZSS7 revealed that non-metabolizable sugars such as L-glucose and mannitol induce mRNA transcription similarly to glucose starvation. When D-glucose or any other metabolizable sugar was supplied, the level of transcripts was reduced. Nucleotide sequence analyses of the six cDNAs allowed identification of five of them by comparison with sequence data bases. The protein encoded by clone pZSS2 is analogous to a wound-induced protein from barley. Clones pZSS4 to pZSS7 encode, respectively, a transmembrane protein, a cysteine protease, a metallothionein-like protein and a chymotrypsin/subtilisin-like protease inhibitor. Clone pZSS3 shares no significant homology with any known sequence.
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    Cloning of a tomato polygalacturonase expressed in abscission (1995)
    Springer
    Plant molecular biology 28 (1995), S. 647-656 
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    ISSN: 1573-5028
    Keywords: abscission ; gene expression ; polygalacturonase ; ethylene ; auxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Abscission, organ separation, is accompanied by cell wall breakdown in separation layer cells. In tomato (Lycopersicon esculentum), ethylene-induced abscission is correlated with an increase in polygalacturonase (PG) and endo-β-1,4-D-glucanase (cellulase) activity. We have identified a putative, abscission-specific cDNA clone for PG, pTAPG1. The TAPG1 cDNA has 43% identity at the amino acid level with the tomato fruit PG. Genomic blot analysis suggests that the gene for TAPG1 is a member of a small subfamily of PG genes that is distinct from the tomato fruit PG. The TAPG1 cDNA hybridizes to mRNA expressed during the course of ethylene-induced leaf and flower abscission. A high level of PG transcript accumulation coincides with the occurrence of abscission. Auxin, an abscission inhibitor, and silver thiosulfate, an ethylene action inhibitor, suppressed accumulation of mRNA in leaf abscission zones complementary to the TAPG1 cDNA. Expression of TAPG1 transcripts is several-fold higher in flower abscission zones than in leaf abscission zones. The identification of cDNAs that encode abscission-specific PG provide and additional tool to study the regulation of abscission and cell wall dissolution in separation layer cells.
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    Intron-dependent transient expression of the maize GapA1 gene (1995)
    Springer
    Plant molecular biology 28 (1995), S. 667-676 
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    ISSN: 1573-5028
    Keywords: gene expression ; promoter ; glyceraldehyde-3-phosphate dehydrogenase ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transient expression experiments show that the maize GapA1 promoter exhibits a requirement for sequences contained within intron 1 and surrounding exon border regions for expression in maize Black Mexican Sweet cells. Maize GapA1-promoter constructs lacking intron 1 are inactive. Intron 1 and its exon border sequences, when reintroduced into constructs lacking introns, restore gene activity whereas intron 2 and its exon borders to not. The minimal promoter so defined encompasses roughly 250 bp upstream of the in vivo transcription start and appears also to include intron 1. An octameric sequence was identified in intron 1 of maize GapA1 which is similar to sequence motifs found in other maize introns known to increase transient expression. Partial restoration of gene expression in GapA1 constructs lacking intron 1 was achieved through insertion of the identified octameric sequence.
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    Aerobic fermentation in tobacco pollen (1995)
    Springer
    Plant molecular biology 28 (1995), S. 739-750 
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    ISSN: 1573-5028
    Keywords: alcohol dehydrogenase ; fermentation ; gene expression ; pollen ; pyruvate decarboxylase ; respiration ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We characterized the genes coding for the two dedicated enzymes of ethanolic fermentation, alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC), and show that they are functional in pollen. Two PDC-encoding genes were isolated, which displayed reciprocal regulation: PDC1 was anaerobically induced in leaves, whereas PDC2 mRNA was absent in leaves, but constitutively present in pollen. A flux through the ethanolic fermentation pathway could be measured in pollen under all tested environmental and developmental conditions. Surprisingly, the major factor influencing the rate of ethanol production was not oxygen availability, but the composition of the incubation medium. Under optimal conditions for pollen tube growth, approximately two-thirds of the carbon consumed was fermented, and ethanol accumulated into the surrounding medium to a concentration exceeding 100 mM.
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    Peroxisomal thiolase mRNA is induced during mango fruit ripening (1995)
    Springer
    Plant molecular biology 28 (1995), S. 811-820 
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    ISSN: 1573-5028
    Keywords: β-oxidation ; gene expression ; fruit ripening ; Mangifera indica ; peroxisomes ; thiolase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fruit ripening is a complex, developmentally regulated process. A series of genes have been isolated from various ripening fruits encoding enzymes mainly involved in ethylene and cell wall metabolism. In order to aid our understanding of the molecular basis of this process in a tropical fruit, a cDNA library was prepared from ripe mango (Mangifera indica L. cv. Manila). By differential screening with RNA poly(A)+ from unripe and ripe mesocarp a number of cDNAs expressing only in ripe fruit have been isolated. This paper reports the characterization of one such cDNA (pTHMF 1) from M. indica which codes for a protein highly homologous to cucumber, rat and human peroxisomal thiolase (EC 2.3.1.16), the catalyst for the last step in the β-oxidation pathway. The cDNA for the peroxisomal mango thiolase is 1305 bp in length and codes for a protein of 432 amino acids with a predicted molecular mass of 45 532 Da. Mango thiolase is highly homologous to cucumber thiolase (80%), the only other plant thiolase whose cloning has been reported, and to rat and human thiolases (55% and 55% respectively). It is shown by northern analysis that during fruit ripening THMF 1 is up-regulated. A similar pattern of expression was detected in tomato fruit. Wounding and pathogen infection do not appear to affect THMF 1 expression. The possible involvement of thiolase in fatty acid metabolism during fruit ripening will be discussed. To our knowledge this is the first report cloning of a plant gene involved in fatty acid metabolism showing an induction during fruit ripening.
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    The chloroplast-located glutamine synthetase of Phaseolus vulgaris L.: nucleotide sequence, expression in different organs and uptake into isolated chloroplasts (1988)
    Springer
    Plant molecular biology 11 (1988), S. 191-202 
    Details
    ISSN: 1573-5028
    Keywords: cDNA sequence ; chloroplast protein import ; gene expression ; glutamine synthetase ; nitrogen metabolism ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Work using a full-length cDNA clone has revealed that the plastid-located glutamine synthetase (GS) of Phaseolus vulgaris is encoded by a single nuclear gene. Nucleotide sequencing has shown that this cDNA is more closely related to a cDNA encoding the plastidic GS of Pisum sativum than to cDNAs encoding three different cytosolic GS subunits of P. vulgaris. The plastid GS subunits are initially synthesized as higher M r (47000) precursors containing an N-terminal presequence of about 50 amino acids which is structurally similar to the presequences of other nuclear-encoded chloroplast proteins. The precursor has been synthesized in vitro and is imported by isolated pea chloroplasts and processed to two polypeptides of the same size as native P. vulgaris chloroplast GS subunits (M r 42000). Experiments with fusion proteins show that the N-terminal 68 amino acids of this precursor allow the cytosolic GS subunit β also to be imported and processed by isolated chloroplasts. Polyadenylated mRNA specifically related to the plastidic GS gene is most highly abundant in chloroplast-containing organs (leaves and stems) but is also detectable in roots and nodules.
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    Molecular basis for the overproduction of 5-enolpyruvylshikimate 3-phosphate synthase in a glyphosate-tolerant cell suspension culture of Corydalis sempervirens (1988)
    Springer
    Plant molecular biology 11 (1988), S. 215-220 
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    ISSN: 1573-5028
    Keywords: glyphosate ; herbicide ; 5-enolpyruvylshikimic acid 3-phosphate (EPSP) synthase ; enzyme overproduction ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell cultures of Corydalis sempervirens adapted to growth in the presence of 5 mM glyphosate [N-(phosphonomethyl)glycine] display a 30- to 40-fold increase in the cellular content of 5-enolpyruvylshikimic acid 3-phosphate (EPSP) synthase, the target enzyme of the herbicide. Translatable mRNA activity as well as transcript levels for EPSP synthase were increased 8-to 12-fold in the adapted (glyphosate-tolerant) as compared to the non-adapted (glyphosate-sensitive) cultures. Northern blot analysis revealed a single 1.8 kb transcript after hybridization with an oligonucleotide probe deduced from the N-terminal amino acid sequence of the enzyme. No significant differences in the relative abundance of EPSP synthase-specific DNA sequences could be detected, however, in Southern and dot blot analyses of restricted DNA isolated from the two cultures. We conclude that the overproduction of EPSP synthase in glyphosate-tolerant C. sempervirens cells is not based on the amplification of the corresponding gene.
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    Cloning and characterization of a cDNA encoding a mRNA rapidly-induced by ABA in barley aleurone layers (1988)
    Springer
    Plant molecular biology 11 (1988), S. 495-506 
    Details
    ISSN: 1573-5028
    Keywords: abscisic acid ; barley aleurone layers ; cDNA cloning ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Abscisic acid (ABA) inhibits the gibberellic acid induced synthesis of α-amylase in barley aleurone layers, yet ABA itself induces more than a dozen polypeptides (Lin & Ho, Plant Physiol 82: 289–297, 1986). As part of our effort to elucidate the molecular action of ABA in barley aleurone layers, we have isolated and characterized an ABA-induced cDNA clone, pHV A1. This cDNA clone hybridizes to an RNA species of approximately 1.1 kb from ABA-treated barley aleurone layers. The level of this mRNA is tripled within 40 minutes after ABA treatment, reaches a peak at 8–12 h, and is present up to 48 h. The induction of this mRNA responds to concentrations of ABA as low as 10-9 M, but higher ABA concentrations induce higher expression of this mRNA. The products of hybrid-select translation and in vitro transcription/translation with pHV A1 comigrate on SDS gel as a 27 kDa polypeptide. However, the sequence of pHV A1 indicates that it has an open reading frame encoding a 22 kDa protein. This size discrepancy is probably due to the high content of the basic amino acid, lysine. This notion has been confirmed by two-dimensional gel electrophoresis showing that this polypeptide is one of the most basic proteins in ABA-treated barley aleurone layers. The deduced amino acid sequence of pHV A1 contains nine imperfect repeats 11 amino acids long which share homology with cotton Lea 7 protein (Baker, Steele & Dure, Plant Mol Biol, in press). The identity and function of the encoded product of pHV A1 is under investigation.
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    Molecular characterization of plastid pyruvate kinase from castor and tobacco (1995)
    Springer
    Plant molecular biology 27 (1995), S. 79-89 
    Details
    ISSN: 1573-5028
    Keywords: pyruvate kinase ; plastid ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Clones encoding two different forms of plastid pyruvate kinase (PKp; EC 2.7.1.40) have been isolated from both castor and tobacco seed cDNA libraries. One form, designated PKpA, from castor was described in a previous report, and the tobacco homologue of PKpA has now been isolated. In addition, a second cDNA, designated PKpG, has been identified and sequenced in both species. Western blot analysis, using antibodies raised against protein overexpressed from these clones, indicates that they encode the two predominant polypeptides of plastid pyruvate kinase from developing castor endosperm. In castor, both PKpA and PKpG are encoded by single genes. In the allotetraploid Nicotiana tabacum, there are two copies of each, one derived from each of the progenitors of this species. The expression of the genes for PKpA and PKpG was examined in various tissues from both castor and tobacco. In castor, both forms are expressed in developing and germinating endosperm and in the root but neither is expressed in the leaf. In tobacco, both forms are expressed in developing seeds but in mature tissues, PKpA is most abundant in roots and PKpG in leaves.
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    The phenylalanine ammonia-lyase gene family in Arabidopsis thaliana (1995)
    Springer
    Plant molecular biology 27 (1995), S. 327-338 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; multi-gene family ; phenylalanine ammonia-lyase ; phenylpropanoids ; promoters ; secondary metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phenylpropanoid derivatives are a complex class of secondary metabolites that have many important roles in plants during normal growth and in responses to environmental stress. Phenylalanine ammonialyase (PAL) catalyzes the first step in the biosynthesis of phenylpropanoids, and is usually encoded by a multi-gene family. Genomic clones for three Arabidopsis thaliana PAL genes containing the entire protein-coding region and upstream and downstream sequences have been obtained and completely sequenced. Two A. thaliana PAL genes (PAL1 and PAL2) are structurally similar to PAL genes that have been cloned from other plant species, with a single intron at a conserved position, and a long highly conserved second exon. Previously identified promoter motifs plus several additional sequence motifs were found in the promoter regions of PAL1 and PAL2. Expression of PAL1 and PAL2 is both qualitatively and quantitatively similar in different plant organs and under various inductive conditions. A third A. thaliana PAL gene, PAL3, differs significantly from PAL1 and PAL2 and other sequenced plant PAL genes. PAL3 contains an additional intron, and its deduced amino acid sequence is less homologous to other PAL proteins. The PAL3 promoter region lacks several sequence motifs conserved between A. thaliana PAL1 and PAL2, as well as motifs described in other genes involved in phenylpropanoid metabolism. A. thaliana PAL3 was expressed at very low levels under the conditions examined.
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    A type I element composed of the hexamer (ACGTCA) and octamer (CGCGGATC) motifs plays a role(s) in meristematic expression of a wheat histone H3 gene in transgenic rice plants (1995)
    Springer
    Plant molecular biology 27 (1995), S. 17-26 
    Details
    ISSN: 1573-5028
    Keywords: cis factor ; gene expression ; promoter ; transgenic rice ; wheat histone H3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Type I element (CCACGTCACCGATCCGCG) is a well-conserved regulatory element found in proximal promoter region of a certain class of plant histone genes, that is composed of two independent cis-acting elements of the hexamer (ACGTCA) and the reverse-oriented octamer (GATCCGCG) motifs. To investigate functional role(s) of the type I element in regulation of a wheat histone H3 gene (TH012) promoter activity in vivo, base substitution mutations were introduced into the element and activities of the mutated promoters were examined in cultured rice cells, and in regenerated roots and anther walls of transgenic rice plants by employing a GUS reporter system. Mutations of each or both of the hexamer and the octamer motifs caused a reduction in the promoter activity in protoplasts transfected transiently or stably transformed calli. The mutation of the octamer motif with or without the mutation of the hexamer motif caused a marked reduction of the promoter activity in the root meristem of transgenic rice although the mutation of the hexamer motif alone caused a weak reduction. In contrast to these results, no effect of the mutations of either the hexamer or the octamer motif was found in the anther wall in which replication-independent activity of the H3 promoter was observed. Our results suggested that the hexamer and the octamer motifs may play important role(s) in regulation of replication-dependent but not of replication-independent expression of the wheat histone H3 gene.
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    URL: http://dx.doi.org/10.1007/BF00019175
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    GASA, a gibberellin-regulated gene family from Arabidopsis thaliana related to the tomato GAST1 gene (1995)
    Springer
    Plant molecular biology 27 (1995), S. 743-752 
    Details
    ISSN: 1573-5028
    Keywords: plant hormone ; gibberellic acid ; GA-responsive ; gene expression ; HCA ; hydrophobic cluster analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A multiple gene family of at least four members, related to a GA-stimulated transcript (GAST1) from tomato, was characterized in Arabidopsis thaliana by analysing four related cDNAs, named GASA1 to GASA4. The corresponding peptides display comparable structural features: (1) a putative signal peptide of 18 to 23 residues; (2) a highly divergent hydrophilic region of about 22 amino acids; (3) a conservative 60 amino acid C-terminal domain containing 12 cysteines. This organization has also bean shown in two related peptides from tomato, GAST1 found in shoots and RSI-1 found in early lateral roots. Southern blot hybridization patterns showed single-copy genes for all four members of the GASA family. Accumulation of the various transcripts, monitored by northern blot hybridization, indicated that the various genes are expressed differentially in plant organs. Specific mRNAs were mostly detected in flower buds and immature siliques in the case of GASA1, in siliques and dry seeds in the case of GASA2 and 3, and in growing roots and flower buds in the case of GASA4. At least two of the GASA genes are activated in GA-deficient mutant ga5, as early as 4 to 8 h after spraying with 50 μM GA3. The complex patterns of expression and regulation of the various genes suggest that the related peptides are involved in a developmental regulation process in Arabidopsis.
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    Molecular cloning of two novel rice cDNA sequences encoding putative calcium-dependent protein kinases (1995)
    Springer
    Plant molecular biology 27 (1995), S. 953-967 
    Details
    ISSN: 1573-5028
    Keywords: calcium-dependent protein kinase ; gene expression ; protein phosphorylation ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated, from a cDNA library constructed from rice coleoptiles, two sequences, OSCPK2 and OSCPK11, that encode for putative calcium-dependent protein kinase (CDPK) proteins. OSCPK2 and OSCPK11 cDNAs are related to SPK, another gene encoding a rice CDPK that is specifically expressed in developing seeds [20]. OSCPK2 and OSCPK11-predicted protein sequences are 533 and 542 amino acids (aa) long with a corresponding molecular mass of 59436 and 61079 Da respectively. Within their polypeptide chain, they all contain those conserved features that define a plant CDPK; kinase catalytic sequences are linked to a calmodulin-like regulatory domain through a junction region. The calmodulin-like regulatory domain of the predicted OSCPK2 protein contains 4 EF-hand calcium-binding sites while OSCPK11 has conserved just one canonical EF-hand motif. In addition, OSCPK2-and OSCPK11-predicted proteins contain, at their N-terminal region preceding the catalytic domain, a stretch of 80 or 74 residues highly rich in hydrophilic amino acids. Comparison of the NH2-terminal sequence of all three rice CDPKs so far identified (OSCPK2, OSCPK11 and SPK) indicates the presence of a conserved MGxxC(S/Q)xxT motif that may define a consensus signal for N-myristoylation. OSCPK2 and OSCPK11 proteins are both encoded by a single-copy gene and their polyadenylated transcripts are 2.4 and 3.5 kb long respectively. OSCPK2 and OSCPK11 mRNAs are equally abundant in rice roots and coleoptiles. A 12 h white light treatment of the coleoptiles reduces the amount of OSCPK2 mRNA with only a slight effect on the level of OSCPK11 transcript. With anoxic treatments, OSCPK2 mRNA level declined significantly and promptly while the amount of OSCPK11 transcript remained constant.
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    Isolation, sequence and expression in Escherichia coli of the nitrite reductase gene from the filamentous, thermophilic cyanobacterium Phormidium laminosum (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1037-1042 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; cyanobacterium ; nitrite reductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nitrite reductase (NiR) gene (nirA) has been isolated and sequenced from the filamentous, thermophilic non-N2-fixing cyanobacterium Phormidium laminosum. Putative promoter-like and Shine-Dalgarno sequences appear at the 5′ end of the 1533 bp long nir-coding region. The deduced amino acid sequence of NiR from P. laminosum corresponds to a 56 kDa polypeptide, a size identical to the molecular mass previously determined for the pure enzyme, and shows a high identity with amino acid sequences from ferredoxin-dependent NiR. This cyanobacterial NiR gene has been efficiently expressed in Escherichia coli DH5α from the E. coli lac promoter and probably from the P. laminosum NiR promoter.
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    Structure of a maize tryptophan synthase alpha subunit gene with pith enhanced expression (1995)
    Springer
    Plant molecular biology 27 (1995), S. 1183-1188 
    Details
    ISSN: 1573-5028
    Keywords: differential screening ; maize ; pith ; trpA ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA derived from an abundant maize pith mRNA transcript and its corresponding genomic equivalent have been isolated and characterized. High transcript levels are seen in the pith and young leaves of maize plants, while no transcript is detected in seed tissue of any age. The protein encoded by the isolated gene has considerable homology with tryptophan synthase alpha subunit (trpA) from other organisms and the cDNA clone can complement an E. coli trpA mutant. These data support the conclusion that this cDNA and the corresponding genomic clone encode a maize trpA protein.
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    Isolation of a novel Arabidopsis ozone-induced cDNA by differential display (1995)
    Springer
    Plant molecular biology 29 (1995), S. 91-98 
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    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; gene expression ; hypersensitive response ; oxidative stress ; ozone ; pathogenesis-related proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have used a reverse transcriptase polymerase chain reaction procedure (differential display) to isolate cDNAs corresponding to transcripts that accumulate in ozone-treated Arabidopsis thaliana. In this report we describe the characterization of an ozone-induced transcript, AtOZI1. AtOZI1 mRNA in untreated plants was detected at low levels in cotyledons, leaves, and flower buds and at higher levels in roots and mature flowers. AtOZI1 mRNA accumulation was transiently induced in leaves 3- to 5-fold within the first 6 h of ozone treatment. AtOZI1 mRNA accumulation was also transiently induced 3- to 6-fold by photopathogenic Pseudomonas strains. Sequence analysis of AtOZI1 revealed that it encodes a 8.6 kDa basic protein that contains a putative signal peptide and two potential phosphorylation sites. Our results suggest that AtOZI1 represents a novel stress-related protein that accumulates in response to the production of active oxygen species.
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    Molecular study of a light-harvesting apoprotein of Giraudyopsis stellifer (Chrysophyceae) (1995)
    Springer
    Plant molecular biology 29 (1995), S. 135-148 
    Details
    ISSN: 1573-5028
    Keywords: chromophyte ; Chrysophyceae ; light-harvesting complex protein gene ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated a gene from a library of nuclear DNA for a chlorophyll a/c-binding protein (named Cac for chl a/c by analogy with Cab for chl a/b) of a chromophyte alga, Giraudyopsis stellifer, and sequenced it. The comparison of the deduced amino acid sequence with other chl a/c-and chl a/b-binding protein sequences shows that structural and functional features, i.e. the arrangement ‘en X’ of the two A and B transmembrane helices and the putative chl a-binding sites, are shared by both Chlorophyta and Chromophyta. Moreover, in contrast to Chlorophyta, a very strong identity is found among Chromophyta in the C helix, suggesting a major function associated to this specific region. Nevertheless, the primary structure of the apoprotein does not seem affected by the pigment composition in Chromophyta. As in the few other examples currently known, we confirm that the cac genes are nuclear-encoded and are part of a multigenic family. Northern blots, performed on poly(A)+ mRNA from G. stellifer, give evidence that the cac gene is light-induced at a transcriptional level and that no expression can be observed in the dark.
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    Isolation and expression pattern of a cDNA encoding a cathepsin B-like protease from Nicotiana rustica (1995)
    Springer
    Plant molecular biology 29 (1995), S. 379-384 
    Details
    ISSN: 1573-5028
    Keywords: cathepsin B ; gene expression ; Nicotiana ; thiol protease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sequence analysis of a 1.33 kb clone from a root cDNA library of Nicotiana rustica revealed an open reading frame encoding a protein of 356 amino acids. The deduced protein has high levels of homology to human cathepsin B protease and a cathepsin B-like cysteine protease from wheat but much lower levels of homology with other plant cysteine proteinases. Southern blotting experiments suggest a limited number of cathepsin B-like genes are present in the genome of N. rustica and also that of N. tabacum. RNA analysis involving a range of tissues, harvested from both Nicotiana species 4–5 h after the beginning of a 16 h photoperiod, revealed the cathepsin B-like gene was being expressed strongly in roots, stem and developing flowers but weakly in mature leaves. Further analysis of RNA extracted from leaf tissue of N. tabacum revealed the gene showed rhythmic expression and also that its expression increased in response to wounding. Analysis of leaf tissues harvested during the latter part of a 16 h photoperiod (11 and 16 h after illumination commenced) showed that transcript levels were two three times higher than in leaf tissue harvested either towards the end of the dark period or 5 h after illumination commenced. When leaf tissue was wounded at 11:00 (5 h after plants were illuminated), and harvested for RNA extraction 6 h later, the level of cathepsin B-like transcript in mesophyll tissue was found to be increased ca. 2-fold relative to the level detected in unwounded controls.
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    Tissue-specific and light-responsive regulation of the promoter region of the Arabidopsis thaliana chloroplast ω-3 fatty acid desaturase gene (FAD7) (1995)
    Springer
    Plant molecular biology 29 (1995), S. 599-609 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; chloroplast ; gene expression ; ω-3 fatty acid desaturase ; promoter ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Arabidopsis FAD7 gene encodes a chloroplast ω-3 fatty acid desaturase that catalyzes the desaturation of lipid-linked dienoic fatty acids (18:2 and 16:2). An 825 bp FAD7 promoter fragment upstream from the transcriptional start point contained several short sequences which were homologous to the cis-elements (box II, G-box, etc.) conserved in many light-responsive genes. We introduced the FAD7 promoter fused to the β-glucuronidase (GUS) or the luciferase (LUC) reporter gene into tobacco plants. The −825 promoter sequence conferred tissue-specific and light-responsive expression to both these reporter genes in transgenic tobacco, indicating that these expressions of the FAD7 gene were regulated mainly at the transcriptional level. Histochemical GUS staining showed that the activity of the FAD7 promoter is restricted to the tissues with chloroplast-containing cells although the staining was noticeably absent in the chloroplast-containing cells associated with vascular systems. The 5′ deletion experiments of the promoter revealed that the −362/ −166 region, containing two putative box II sequences, was responsible for the tissue-specific and light-responsive expression of the FAD7 gene.
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    Differential regulation of polygalacturonase and pectin methylesterase gene expression during and after heat stress in ripening tomato (Lycopersicon esculentum Mill.) fruits (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1101-1110 
    Details
    ISSN: 1573-5028
    Keywords: tomato ; polygalacturonase ; pectin methylesterase ; heat stress ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of extended heat stress on polygalacturonase (PG; EC 3.2.1.15) and pectin methylesterase (PME; EC 3.1.1.11) gene expression at mRNA, protein and activity levels in ripening tomato fruits were investigated. Steady state levels of PG mRNA declined at temperatures of 27°C and above, and a marked reduction in PG protein and activity was observed at temperatures of 32°C and above. Exogenous ethylene treatment did not reverse heat stress-induced inhibition of PG gene expression. Transfer of heat-stressed fruits to 20°C partly restored PG mRNA accumulation, but the rate of PG mRNA accumulation declined exponentially with duration of heat stress. Heat stress-induced inhibition of PME mRNA accumulation was recoverable even after 14 days of heat stress. In fruits held at 34°C, both PG and PME protein and activity continued to accumulate for about 4 days, but thereafter PG protein and activity declined while little change was observed in PME protein and activity. In spite of increases in mRNA levels of both PG and PME during the recovery of heat-stressed fruit at 20°C, levels of PG protein and activity declined in fruits heat-stressed for four or more days while PME protein and activity levels remained unchanged. Collectively, these data suggest that PG gene expression is being gradually and irreversibly shut off during heat stress, while PME gene expression is much less sensitive to heat stress.
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    The phytochrome gene family in tomato includes a novel subfamily (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1143-1155 
    Details
    ISSN: 1573-5028
    Keywords: gene family ; gene expression ; photoreceptor ; phytochrome ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Data presented here define five tomato phytochrome genes (PHY) and indicate the existence of additional PHY in the tomato genome. Portions of each gene, encoding amino acids 203 through 315 in a consensus amino acid sequence, were amplified by polymerase chain reaction. Four of these genes, PHYA, PHYB1, PHYB2 and PHYE, are members of previously identified PHY subfamilies, while the fifth, PHYF, is identified as a member of a new PHY subfamily. PHYA, PHYB1, PHYB2 and PHYE fragments encode amino acid sequences that share 88% to 98% sequence identity with their Arabidopsis counterparts. The PHYF fragment, however, encodes a polypeptide that shares only 65% to 74% sequence identity with previously identified Arabidopsis phytochromes. A phylogenetic analysis suggests that PHYF arose soon after, or perhaps prior to, the origin of angiosperms. This analysis leads to the prediction that PHYF might be widespread among angiosperms, including both monocotyledons and dicotyledons. Each of the five tomato PHY is expressed as a transcript of sufficient size to encode a full-length phytochrome apoprotein. Two PHYF transcripts, 4.4 and 4.7 kb in length, have been detected in 9-day-old light-grown seedlings, consistent with either multiple transcription start sites or differential processing. Analyses of genomic Southern blots hybridized with radiolabelled RNA probes derived from the five tomato PHY, as well as Arabidopsis PHYC, indicate that the tomato genome contains as many as 9 to 13 PHY. The tomato PHY family is apparently not only different from, but also larger than, the PHY family presently described for Arabidopsis.
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    The AQP2 water channel: Effect of vasopressin treatment, microtubule disruption, and distribution in neonatal rats (1995)
    Springer
    The journal of membrane biology 143 (1995), S. 165-175 
    Details
    ISSN: 1432-1424
    Keywords: Water channels ; Vasopressin ; Rat kidney ; Immunocytochemistry ; Microtubules ; Cell polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aquaporin 2 is a collecting duct water channel that is located in apical vesicles and in the apical plasma membrane of collecting duct principal cells. It shares 42% identity with the proximal tubule/thin descending limb water channel, CHIP28. The present study was aimed at addressing three questions concerning the location and behavior of the AQP2 protein under different conditions. First, does the AQP2 channel relocate to the apical membrane after vasopressin treatment? Our results show that AQP2 is diffusely distributed in cytoplasmic vesicles in collecting duct principal cells of homozygous Brattleboro rats that lack vasopressin. In rats injected with exogenous vasopressin, however, AQP2 became concentrated in the apical plasma membrane of principal cells, as determined by immunofluorescence and immunogold electron microscopy. This behavior is consistent with the idea that AQP2 is the vasopressin-sensitive water channel. Second, is the cellular location of AQP2 modified by microtubule disruption? In normal rats, AQP2 has a mainly apical and subapical location in principal cells, but in colchicine-treated rats, it is distributed on vesicles that are scattered throughout the entire cytoplasm. This is consistent with the dependence on microtubules of apical protein targeting in many cell types, and explains the inhibitory effect of microtubule disruption on the hydroosmotic response to vasopressin in sensitive epithelia, including the collecting duct. Third, is AQP2 present in neonatal rat kidneys? We show that AQP2 is abundant in principal cells from neonatal rats at all days after birth. The detection of AQP2 in early neonatal kidneys indicates that a lack of this protein is not responsible for the relatively weak urinary concentrating response to vasopressin seen in neonatal rats.
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    Electron microscopic immunocytochemical demonstration of separate neurophysin-vasopressinergic and neurophysin-oxytocinergic nerve fibres in the neural lobe of the rat hypophysis (1976)
    Springer
    Cell & tissue research 171 (1976), S. 31-37 
    Details
    ISSN: 1432-0878
    Keywords: Rat posterior pituitary ; Neurophysin-vasopressinergic and neurophysin-oxytocinergic fibres ; Immunocytochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of the unlabeled antibody peroxidase-antiperoxidase (PAP) technique at the electron microscopic level, it was demonstrated that the hormones of the neural lobe of the rat hypophysis are located in separate neurophysin-vasopressinergic and neurophysin-oxytocinergic nerve fibres. These observations confirm the results of our previous immunocytochemical studies at the light microscopic level.
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    Quantitative autoradiographic assessment of 3H-estradiol uptake in immunocytochemically characterized pituitary cells (1976)
    Springer
    Cell & tissue research 166 (1976), S. 25-35 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Cell types ; Estrogen ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Dry-mount autoradiography was combined with peroxidase immunocytochemistry to examine estrogen uptake in four pituitary cell types. Quantification by silver grain counts was used to compare 3H-estradiol uptake in nuclei of pituitary cells 60 min after i.v. injection into short-term (control) and long-term ovariectomized and in long-term thyroidectomized rats. Under all three hormonal states, the order of labeling intensity was: gonadotropes 〉 somatotropes 〉 lactotropes 〉 thyrotropes. Long-term ovariectomy caused a significant increase in estrogen uptake of gonadotropes, somatotropes and lactotropes, while uptake in thyrotropes decreased. Long-term thyroidectomy decreased uptake in somatotropes, lactotropes and thyrotropes while gonadotropes remained unchanged.
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    Immunocytochemical localization of nerve growth factor (NGF) in the submandibular gland of adult mice by light and electron microscopy (1976)
    Springer
    Cell & tissue research 169 (1976), S. 289-299 
    Details
    ISSN: 1432-0878
    Keywords: Nerve growth factor ; Submandibular gland mice ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nerve growth factor (NGF) was localized in the submandibular gland of adult male mice by a direct immunocytochemical method using highly purified antibodies against NGF coupled to horseradish peroxidase. In light microscopic sections the reaction product was entirely confined to the cells of the secretory tubules. The acinar part of the gland was free of reaction product. This finding was confirmed by electron microscopy. Within the cells NGF was localized exclusively in the apical secretory granules. No reaction was observed in the rough endoplasmic reticulum, the Golgi region or in the granules of the basal part of the cells. This observation favours the assumption that NGF is derived from a precursor molecule and that the precursor is transformed into immunologically active NGF within the secretory granules during their transport from the basal to the apical part of the tubular cells. Stimulation of the submandibular gland with carbachol (2 mg/kg) led to a massive release of the content of the secretory granules, including NGF, into the salivary duct.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219602
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    Ultrastructural immunocytochemical localization of luteinizing hormone-releasing hormone (LH-RH) in the median eminence of the guinea pig (1976)
    Springer
    Cell & tissue research 169 (1976), S. 157-166 
    Details
    ISSN: 1432-0878
    Keywords: Luteinizing hormone-releasing hormone ; Median eminence ; Electron microscopy ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary LH-RH was localized at the ultrastructural level in axons and nerve terminals of the median eminence of the male guinea pig. LH-RH positive neuronal profiles were most concentrated in the medial-dorsal aspect of the infundibular stalk and in the post-infundibular median eminence at the level immediately following separation of the stalk from the base of the brain. LH-RH containing axon profiles were most abundant in the palisade zone; nerve terminals in contact with the hypophysial portal vasculature were relatively rare. The hormone was present within granules that measured 900–1,200 Å in axons of the palisade zone and 400–800 Å in nerve terminals abutting on the portal plexus. The differently sized granules represent heterogeneous populations.
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    URL: http://dx.doi.org/10.1007/BF00214205
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    Cell differentiation of the fetal rat anterior pituitary in vitro (1976)
    Springer
    Cell & tissue research 170 (1976), S. 263-273 
    Details
    ISSN: 1432-0878
    Keywords: Fetal pituitary ; Cell differentiation ; Immunocytochemistry ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Adenohypophysial primordia of rat embryos at 13 to 15 days gestation were cultured in Parker 199 synthetic medium for 2 to 11 days. At the end of the culture period their fine structure and the presence of immunoreactive trophic hormones using the peroxidase-labeled antibody technique were investigated. The degree of differentiation in the glands depends largely on the age of the embryos furnishing the explants. Cultured pituitaries explanted on the 13th day of gestation contain only ACTH-positive cells and about 15% of the cells are granular. The granules are 50–100 nm in diameter in some cells, while in other cells they range from 50 to 200 nm. In cultivated adenohypophysial primordia of embryos on the 15th day of intrauterine life ACTH, prolactin, LH and TSH cells are evident, but only the same two kinds of granular cells can be observed with the electron microscope. The extent of cytodifferentiation in the glands explanted on the 14th day of gestation is intermediate between the two other groups. The data suggest that the fetal rat pituitary has the capacity of self-differentiation but to a lesser extent than that of the in situ hypophysis.
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    URL: http://dx.doi.org/10.1007/BF00224303
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    Immunocytochemical demonstration of separate vasotocinergic and mesotocinergic neurons in the amphibian hypothalamic magnocellular neurosecretory system (1976)
    Springer
    Cell & tissue research 175 (1976), S. 289-296 
    Details
    ISSN: 1432-0878
    Keywords: Amphibian hypothalamus ; Vasotocinergic and mesotocinergic neurons ; Neurophysins ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the unlabeled antibody peroxidase-antiperoxidase (PAP) technique at the light microscopic level, it was demonstrated that, in the amphibian magnocellular hypothalamo-hypophysial neurosecretory system, vasotocin and mesotocin are synthesized in separate neurons. A tendency to preferential location of the two kinds of neuronal perikarya is described. The neurosecretory perikarya are the origin of separate vasotocinergic and mesotocinergic axons. In the neural lobe, the pattern of distribution of the two types of axons is different. The coarse ventricular “dendrites” of both kinds of neurons are hormone-containing processes. Staining with anti-bovine neurophysin I serum suggested that the vasotocinergic and the mesotocinergic neurons synthesize different neurophysins.
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    URL: http://dx.doi.org/10.1007/BF00218707
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    Electron microscopic immunocytochemical demonstration of separate vasotocinergic and mesotocinergic nerve fibres in the neural lobe of the amphibian hypophysis (1976)
    Springer
    Cell & tissue research 173 (1976), S. 461-464 
    Details
    ISSN: 1432-0878
    Keywords: Amphibian posterior pituitary ; Vasotocinergic and mesotocinergic fibres ; Immunocytochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the unlabeled antibody peroxidase-antiperoxidase (PAP) technique at the electron microscopic level, it was demonstrated that the hormones of the posterior pituitary of Rana temporaria are located in separate vasotocinergic and mesotocinergic nerve fibres. This observation confirms the results of our previous immunocytochemical studies at the light microscopic level.
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    URL: http://dx.doi.org/10.1007/BF00224308
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    Immunocytochemical study of the hypothalamo-neurohypophysial system (1976)
    Springer
    Cell & tissue research 175 (1976), S. 165-181 
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    ISSN: 1432-0878
    Keywords: Pig ; Neurophysin-I ; Neurophysin-II ; Immunocytochemistry ; Specific neurophysin neurosecretory systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antibodies raised against porcine neurophysin-I and porcine neurophysin-II using an injection regime in rabbits over a short time period, were used to localize neurophysin-I and neurophysin-II in hypothalamic neurosecretory elements of the domestic pig. In transverse section, neurophysin-II containing cells were more abundant in the dorsal medial region of the rostral supraoptic nucleus (SON) as compared with the distribution of neurophysin-I neurons. The main bulk of the cells of the SON were heavily stained for neurophysin-I with neurophysin-II containing cells positioned dorsal from the edge of the optic chiasma. Neurosecretory cells of the SON as seen in sagittal section also showed a differential staining for neurophysins-I and -II. Rostral regions of the pig paraventricular nucleus (PVN) contained magnocellular elements near the third ventricle which were stained predominantly for neurophysin-II. In regions corresponding to the caudal PVN there appeared two populations of neurosecretory neurons: (a) an area of cells adjacent to the third ventricle which contained neurophysin-II antigen and (b) a group of densely populated cells in the dorsal-lateral region which was stained for neurophysin-I. The results support the existence in the pig of at least two distinct populations of neurosecretory neurons corresponding to the neurophysin-I and neurophysin-II neurosecretory system.
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    URL: http://dx.doi.org/10.1007/BF00232077
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    Mutant vasopressin precursor in the endoplasmic reticulum of the Brattleboro rat. Ultrastructural evidence from individual “vasopressin” cells localized with the light microscope by use of a new gold/silver method for immunostain enhancement (1988)
    Springer
    Cell & tissue research 253 (1988), S. 671-676 
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    ISSN: 1432-0878
    Keywords: Vasopressin precursor ; Endoplasmic reticulum ; Gold/silver intensification ; Immunocytochemistry ; Brattleboro rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This ultrastructural study demonstrates that the vasopressin immunoreactivity found in the occasional, densely stained cells in the hypothalamus of the homozygous Brattleboro rat is localized in the rough endoplasmic reticulum. 50-μm Vibratome sections were stained with anti-vasopressin serum by use of a peroxidase method with 3,3-diaminobenzidine as chromogen. The diaminobenzidine end-product has a specific capability to bind gold particles from a chloroauric acid solution and the bound gold was used to precipitate silver grains from a silver developer. The stained sections were flat embedded in resin and ultrathin sections were cut of areas containing the immuno-identified occasional cells. In these densely stained, vasopressin-immunoreactive cells of homozygous Brattleboro rats the rough endoplasmic reticulum was dilated. The lumen of the reticulum contained both end-products of diaminobenzidine and gold/silver grains, but some parts of the reticulum appeared unstained. No other cell organelles were immunostained and no secretory granules were found. In control rats, gold/silver deposits were found throughout the cytoplasm of vasopressin-immunoreactive cells. In these immunostained cells secretory granules were seen.
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    URL: http://dx.doi.org/10.1007/BF00219759
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    Lysosomes and pancreatic islet function (1988)
    Springer
    Cell & tissue research 252 (1988), S. 9-15 
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    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Insulin ; Immunocytochemistry ; Lysosomes ; Crinophagy ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrastructural studies of pancreatic islets have suggested that crinophagy provides a possible mechanism for intracellular degradation of insulin in the insulin-producing B-cells. In the present study, a quantitative estimation of crinophagy in mouse pancreatic islets was attempted by morphometric analysis of lysosomes containing immunoreactive insulin. Isolated islets were incubated in tissue culture for one week in 3.3, 5.5 or 28 mmol/l glucose. The lysosomes of the pancreatic B-cells were identified by morphological and enzyme-cytochemical criteria and divided into three subpopulations comprising primary lysosomes and insulin-positive or insulin-negative secondary lysosomes. Both the volume and numerical density of the primary lysosomes increased with increasing glucose concentration. The proportion of insulin-containing secondary lysosomes was highest at 5.5 and lowest at 3.3 mmol/l glucose. Insulin-negative secondary lysosomes predominated at 3.3 mmol/l glucose. Studies of the dose-response relationships of glucose-stimulated insulin biosynthesis and insulin secretion of the pancreatic islets showed that biosynthesis had an apparent Km-value for glucose of 7.0 mmol/l, whereas it was 14.5 mmol/l for secretion. The pronounced crinophagic activity at 5.5 mmol/l glucose may thus be explained by the difference in glucose sensitivity between insulin biosynthesis and secretion resulting in an intracellular accumulation of insulin-containing secretory granules. The predominance of insulin-negative secondary lysosomes at 3.3 mmol/l glucose may reflect an increased autophagy, whereas the predominance of primary lysosomes at 28 mmol/l glucose may reflect a generally low activity of intracellular degradative processes.
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    URL: http://dx.doi.org/10.1007/BF00213820
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    Structural studies of nerve terminals containing melanin-concentrating hormone in the eel, Anguilla anguilla (1988)
    Springer
    Cell & tissue research 251 (1988), S. 433-439 
    Details
    ISSN: 1432-0878
    Keywords: Melanin-concentrating hormone ; Immunocytochemistry ; Pituitary gland ; White- and black-background adaptation ; Teleost, Anguilla anguilla
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Eels were adapted to black- or white-coloured backgrounds and the pituitary glands were prepared for light and electron microscopy. Immunocytochemical staining was used to study the distribution of the neurohypophysial melanin-concentrating hormone in the neurointermediate lobe. The hormone was located in small, elliptical, electron-opaque neurosecretory granules, measuring approximately 120×90 nm. The neurones terminated on blood vessels in the centre of the neurohypophysis and on the basement membrane separating neural and intermediate lobe tissues. The results of both light and electron immunocytochemistry and of radioimmunoassay are consistent with a higher rate of hormone release from eels adapted to white backgrounds than from those adapted to black backgrounds. In addition to this, when fish that had been adapted to white tanks were transferred to black tanks, there was an accumulation of irMCH in the gland and an increased numerical density of secretory granules at nerve terminals. These results reinforce the proposal that MCH is released during adaptation to a white background, to cause melanin concentration and to inhibit MSH release, and that its release is halted in black-adapted fish.
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    URL: http://dx.doi.org/10.1007/BF00215852
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    Localization of pigment-dispersing hormone (PDH) immunoreactivity in the central nervous system of Carcinus maenas and Orconectes limosus (Crustacea), with reference to FMRFamide immunoreactivity in O. limosus (1988)
    Springer
    Cell & tissue research 253 (1988), S. 199-208 
    Details
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone ; FMRFamide ; Immunocytochemistry ; Carcinus maenas ; Orconectes limosus (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of antisera raised against synthetic pigment-dispersing hormone (PDH) of Uca pugilator and FMRFamide, the distribution of immunoreactive structures in the central nervous system (CNS) of Carcinus maenas and Orconectes limosus was studied by light microscopy. In both species, a total of 10–12 PDH-positive perikarya occur amongst the anterior medial, dorsal lateral and angular somata of the cerebral ganglion (CG). In C. maenas, one PDH-perikaryon was found in each commissural ganglion (COG) and several more in the thoracic ganglion. In O. limosus, only four immunopositive perikarya could be demonstrated in the ventral nerve cord, i.e., two somata in the anterior and two in the posterior region of the suboesophageal ganglion (SOG). PDH-immunoreactive tracts and fiber plexuses were present in all central ganglia of both species, and individual axons were observed in the connectives. FMRFamide-immunoreactivity was studied in O. limosus only. Neurons of different morphological types were found throughout the entire CNS, including numerous perikarya in the anterior medial, anterior olfactory, dorsal lateral and posterior cell groups of the CG. Four perikarya were found in the COG, six large and numerous smaller ones in the SOG, and up to eight cells in each of the thoracic and abdominal ganglia. In each ganglion, the perikarya form fiber plexuses. Axons from neurons belonging to the CG could be traced into the ventral nerve cord; nerve fibers arising from perikarya in the SOG appeared to project to the posterior ganglia. In none of the structures examined colocalization of PDH- and FMRF-amide-immunoreactivity was observed.
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    URL: http://dx.doi.org/10.1007/BF00221755
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    Choline acetyltransferase immunocytochemical staining of the rat supraoptic nucleus and its surroundings (1988)
    Springer
    Cell & tissue research 254 (1988), S. 119-124 
    Details
    ISSN: 1432-0878
    Keywords: Choline acetyltransferase ; Immunocytochemistry ; Light and electron microscopy ; Supraoptic nucleus ; Paraventricular nucleus ; Rat (Wistar, Long Evans, Brattleboro)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two different monoclonal antibodies raised against choline acetyltransferase were used, together with preembedding immunocytochemical techniques, to visualize the possible cholinergic innervation of the supraoptic and paraventricular nuclei of the rat hypothalamus. Light microscopy confirmed the presence of a group of bipolar and multipolar immunoreactive neurones in the hypothalamus dorsolateral to the supraoptic nucleus as well as numerous immunopositive fibers. Electron microscopy showed that the immunopositive cell bodies contained the usual perikaryal organelles while most immunoreactive fibers appeared dendritic; immunonegative terminals made synaptic contact onto these profiles. Immunopositive terminals making synaptic contact onto dendritic profiles were also noted in this area. In contrast, light microscopy showed no immunoreactivity to choline acetyltransferase in the magnocellular nuclei themselves. Electron microscopy revealed some immunopositive profiles along the boundaries of both nuclei, along the optic chiasm adjacent to the supraoptic nucleus and in the ventral glial lamina but not within the nuclei proper. Surprisingly, these immunopositive profiles appeared dendritic and were often contacted by one or more immunonegative synapses. Our observations thus indicate that cell bodies and dendrites in the supraoptic and paraventricular nuclei are not directly innervated by cholinergic synapses. The functional significance of the putative cholinergic dendrites in close proximity to magnocellular neurones remains to be determined.
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    URL: http://dx.doi.org/10.1007/BF00220024
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    Presence of an oxytocin-like peptide in the hypothalamus and neurohypophysis of a turtle (Mauremys caspica) and a snake (Natrix maura) (1995)
    Springer
    Cell & tissue research 279 (1995), S. 75-84 
    Details
    ISSN: 1432-0878
    Keywords: Oxytocin ; Neurophysin ; Vasotocin ; Mesotocin ; Suprachiasmatic nucleus ; Medial nucleus of the infundibular recess ; Immunocytochemistry ; Natrix maura (Serpentes) ; Mauremys caspica (Chelonia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The probable presence of oxytocin in the hypothalamo-hypophysial system of two reptilian species, the snake Natrix maura and the turtle Mauremys caspica, was re-investigated. A high-pressure liquid chromatographic analysis of the turtle neural lobe revealed the existence of vasotocin, mesotocin, and a third compound co-eluting with oxytocin. Brains from both species were fixed by vascular perfusion with Bouin's fluid. Adjacent paraffin sections were immunostained using antisera against the following substances: (1) bovine oxytocin-neurophysin; (2) a mixture of bovine oxytocin-neurophysin and vasopressin-neurophysin; (3) dogfish neurophysins; (4) oxytocin; (5) arginine-vasotocin; (6) mesotocin; (7) somatostatin. Immunoreactivity against oxytocin was found in parvocellular neurons of the snake suprachiasmatic nucleus and cerebrospinal-fluid contacting neurons of the medial nucleus of the infundibular recess of both species, the latter immunoreactivity being much more conspicuous in the turtle. Numerous fibers containing immunoreactive oxytocin extended between the medial nucleus of the infundibular recess, and the internal region of the medium eminence and the neural lobe. The oxytocin-immunoreactivity in all locations was completely abolished by preabsorption of the anti-oxytocin serum with three different oxytocin preparations. None of the neurons of the suprachiasmatic and medial nucleus of the infundibular recess, including the oxytocin-immunoreactive elements, reacted with either the antineurophysin sera used, or the anti-vasotocin or anti-mesotocin antibodies. The possible existence of a reptilian oxytocin-neurophysin is discussed. The alternative that, in the reptilian hypothalamus, neurons synthesize a compound closely related to, but different from oxytocin is also considered.
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    URL: http://dx.doi.org/10.1007/BF00300693
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    The biased intracellular accumulation of the β-subunit of salmon gonadotropin (GTH II) in the pituitary of rainbow trout Oncorhynchus mykiss, during gametogenesis (1995)
    Springer
    Cell & tissue research 279 (1995), S. 93-99 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Gonadotropin ; Subunits ; Gonadotropes ; Immunocytochemistry ; Immunoblotting ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone (α and IIβ subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the α- and IIβ-subunits shows that colocalization of the α- and IIβ-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, α-immunolabeling is absent in the large globules, even though IIβ labeling is abundant throughout the period of seasonal gametogenesis. The α-specific antiserum recognizes the intact α-subunit as well as the reduced and deglycosylated α-subunits by immunoblotting. These results indicate that an accumulation of the IIβ-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of IIβ-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained IIβ-subunits remains unknown.
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    URL: http://dx.doi.org/10.1007/BF00300695
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    Regeneration and post-metamorphic development of the central nervous system in the protochordate Ciona intestinalis: a study with monoclonal antibodies (1995)
    Springer
    Cell & tissue research 279 (1995), S. 421-432 
    Details
    ISSN: 1432-0878
    Keywords: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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    URL: http://dx.doi.org/10.1007/BF00318500
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    Presence of embryonic myosin in normal postural muscles of the adult rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 541-548 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Musle ; striated ; skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 μ2), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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    URL: http://dx.doi.org/10.1007/BF00318358
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    Development of melanin-concentrating hormone-immunoreactive elements in the brain of gilthead seabream (Sparus auratus) (1995)
    Springer
    Cell & tissue research 282 (1995), S. 523-526 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Melanin-concentrating hormone ; Immunocytochemistry ; Development ; ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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    URL: http://dx.doi.org/10.1007/s004410050504
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    Ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the dogfish (1995)
    Springer
    Cell & tissue research 282 (1995), S. 33-40 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development ; ontogenetic ; Vitellointestinal duct ; Pancreas ; exocrine ; Pancreas ; endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050456
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    Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium (1995)
    Springer
    Cell & tissue research 281 (1995), S. 77-83 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Esophagus ; Epithelial cells ; Intestinal lectin ; L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells form a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00307960
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    Immunocytochemical localization of CCAP, a novel crustacean cardioactive peptide, in the nervous system of the shore crab, Carcinus maenas L. (1988)
    Springer
    Cell & tissue research 254 (1988), S. 347-360 
    Details
    ISSN: 1432-0878
    Keywords: Crustacean cardioactive peptide (CCAP) ; Proctolin ; FMRFamide ; Leu-enkephalin ; Immunocytochemistry ; Ultrastructural immunogold-labeling ; Pericardial organs ; Neurosecretion ; Carcinus maenas (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Polyclonal antibodies were raised in rabbits against synthetic crustacean cardioactive peptide (CCAP) conjugated to bovine thyroglobulin, and were used to map CCAP-immunoreactive structures in the central nervous system of Carcinus maenas. As expected, the neurohemal pericardial organs (PO) displayed abundant immunoreactivity in nerve fibers and terminals. In addition, immunoreactive neurons were demonstrated in other parts of the nervous system. At least some of them do not appear to terminate in neurohemal structures and may have a non-endocrine, as yet unknown function. Immunoreactive perikarya with a diameter of 25–30 μm occur in the brain. They project into the optic and antennary neuropil, and into the eyestalk. One cell was found in the medulla terminalis of the eyestalk and in the connective ganglion, respectively. From the latter, axonal branches could be traced into the brain and the thoracic ganglia (TG). In the TG, small-diameter perikarya give rise to extensive networks of varicose fibers. Some of the perikarya occur in a characteristic paired arrangement with larger CCAP-immunoreactive somata (diameter 40–50 μm). These pairs of one small and one large cell occur in all mouthpart and leg segments of the TG, except the abdominal ganglia (AG), where only large cells were found. The main projections of the large neurons comprise one or more fibers in each of the seven segmental nerves (SN), leading to neurosecretory terminals in the PO. The fibers in the SN are joined by branches of an ascending axonal tract from the large perikarya in the AG. The large-type perikarya are considered to be the principal source of CCAP in the PO. The optic ganglia in the eyestalk, except the medulla terminalis, the neurohemal sinus gland and the stomatogastric nervous system are devoid of CCAP-immunoreactivity. In axon terminals of the PO, CCAP is not colocalized with other PO-neuropeptides, i.e. proctolin-, FMRFamide-like, and Leu-enkephalin-like immunoreactive materials. Electron-microscopic immunocytochemistry revealed a distinct CCAP-containing granule type in specific axon profiles and terminals in the PO. The architecture of CCAP-immunoreactive neurons is discussed with respect to previous morphological studies on the origin and pathways of fibers terminating in the PO.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00225807
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  • 91
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    Co-existence and origin of peptidergic and adrenergic nerves in the guinea pig uterus (1988)
    Springer
    Cell & tissue research 254 (1988), S. 517-530 
    Details
    ISSN: 1432-0878
    Keywords: Uterus ; Autonomic innervation ; Neuropeptides ; Immunocytochemistry ; Retrograde tracing ; Pregnancy ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The occurrence and distribution of peptidergic nerves in the guinea pig uterus was studied by means of immunocytochemistry using numerous neuropeptide antisera. Neuropeptide Y (NPY)-immunoreactive (IR) nerves were the most abundant, whereas substance P (SP)-, calcitonine gene-related peptide (CGRP)-, and neurokinin A (NKA)-IR nerves were less frequent, and peptide histidine isoleucine (PHI)-IR nerves were the most sparse. Chemical sympathectomy by means of 6-hydroxydopamine, and capsaicin treatment revealed the division of the peptidergic nerves into three separate populations: (1) NPY-IR nerves, which co-existed with adrenergic nerves, (2) SP-, CGRP-and NKA-IR nerves, which mutually co-existed, and (3) PHI-IR nerves. Parallel-running adrenergic/NPY-IR and SP-IR nerves could be found with very similar although not completely identical morphological appearance. Paracervical ganglia contained neurotensin-and dynorphin A-IR cell bodies in addition to cell bodies with immunoreactivities similar to those in prevertebral ganglia. Combined retrograde tracing with True blue and immunocytochemistry showed that the adrenergic and NPY-IR uterine nerves originate in paracervical and prevertebral ganglia. In the prevertebral ganglia the cellular origin was the same for adrenergic and NPY-IR nerves. In contrast, SP-, CGRP-,and NKA-IR nerves originated in dorsal root ganglia. At full-term pregnancy all the neuropeptide immunoreactivities had vanished, probably reflecting a fetus-induced general nerve degeneration.
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    URL: http://dx.doi.org/10.1007/BF00226501
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  • 92
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    Pinealocytes immunoreactive with antisera against secretory glycoproteins of the subcommissural organ: A comparative study (1988)
    Springer
    Cell & tissue research 254 (1988), S. 469-480 
    Details
    ISSN: 1432-0878
    Keywords: Pineal complex ; Pinealocytes, receptor line ; Subcommissural organ ; Immunocytochemistry ; Protein secretion ; Neuroendocrine system Geotria australis (Cyclostomata) ; Onkorhynchus kisutch (Teleostei) ; Eupsophus roseus (Anura) ; Heloderma suspectum, Varanus monitor (Lacertilia) ; Domestic fowl ; Rat ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of light-microscopic immunocyto-chemistry two polyclonal antibodies (AFRU, ASO; see p. 470) directed against secretory glycoproteins of the subcom-missural organ were shown to cross-react with cells in the pineal organ of lamprey larvae, coho salmon, a toad, two species of lizards, domestic fowl, albino rat and bovine (taxonomic details, see below). The AFRU-immunoreactive cells were identified as pinealocytes of the receptor line (pineal photoreceptors, modified photoreceptors or classical pinealocytes, respectively) either due to their characteristic structural features or by combining AFRU-immunoreaction with S-antigen and opsin immunocytochemistry in the same or adjacent sections. Depending on the species, AFRU- or ASO-immunoreactions were found in the entire perikaryon, inner segments, perinuclear area, and in basal processes facing capillaries or the basal lamina. In most cases, only certain populations of pinealocytes were immunolabeled; these cells were arranged in a peculiar topographical pattern. In lamprey larvae, immunoreactive pinealocytes were observed only in the pineal organ, but not in the parapineal organ. In coho salmon, the immunoreaction occurred in S-antigen-positive pinealocytes of the pineal end-vesicle, but was absent from S-antigen-immunoreactive pinealocytes of the stalk region. In the rat, AFRU-immunoreaction was restricted to S-antigen-immunoreactive pinealocytes found in the deep portion of the pineal organ and the habenular region. These findings support the concept that several types of pinealocytes exist, which differ in their molecular, biochemical and functional features. They also indicate the possibility that the AFRU- and ASO-immunoreactive material found in certain pinealocytes might represent a proteinaceous or peptidic compound, which is synthesized and released from a specialized type of pinealocyte in a hormone-like fashion. This cell type may share functional characteristics with peptidergic neurons or paraneurons.
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    URL: http://dx.doi.org/10.1007/BF00226496
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    Mapping of enkephalin-related peptides in the nervous system of the blowfly, Calliphora vomitoria, and their co-localization with cholecystokinin (CCK)- and pancreatic polypeptide (PP)-like peptides (1988)
    Springer
    Cell & tissue research 251 (1988), S. 399-415 
    Details
    ISSN: 1432-0878
    Keywords: Enkephalin-related peptides ; Immunocytochemistry ; Neuropeptides ; Co-existence of peptides ; Neurosecretory cells ; Blowly, Calliphora vomitoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of enkephalin-like immunoreactive material has been studied in the CNS of C. vomitoria. The presence of both Met- and Leu-enkephalin-related peptides is suggested by differential immunostaining with a variety of antisera. Comparisons made between certain of the enkephalin-immunoreactive perikarya, nerve fibres and terminals with cells in corresponding positions as evidenced in previously published neuroanatomical studies of the dipteran brain have suggested specific enkephalinergic pathways. As examples, one Met-enkephalin-immunoreactive neuron appears to link the lobula with the dorsal protocerebrum, and a group of Leu-enkephalin cells in the pars intercerebralis appear to have arborisations in both the central body (fan-shaped body) and the tritocerebral neuropil around the oesophageal foramen. Neuronal pathways of this type indicate that the enkephalin-like peptides of the fly brain are functioning as neurotransmitters and/or neuromodulators. In the thoracic ganglia, symmetrically arranged cells, immunoreactive to both Met- and Leu-enkephalin antisera, are positioned ventrally in pairs on either side of the mid-line in a sagittal plane. Very little immunoreactive material is observed in the neuropil, however, and the source of the accumulation of Leu-enkephalin-immunoreactivity in the dorsal neural sheath is not certain. It is suggested that this material, in contrast to that present in areas of the brain, acts as a neurohormone and that it may have a physiological role following its release into the haemolymph. The enkephalin-like immunoreactive material of certain neurons identified within the brain and thoracic ganglion shows a complex pattern of co-existence with pancreatic polypeptide- and gastrin/cholecystokinin-like peptides.
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    URL: http://dx.doi.org/10.1007/BF00215849
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    Thyrotropin-releasing hormone (TRH)-immunoreactive structures in the brain of the domestic mallard (1988)
    Springer
    Cell & tissue research 251 (1988), S. 441-449 
    Details
    ISSN: 1432-0878
    Keywords: Thyrotropin-releasing hormone (TRH) ; Immunocytochemistry ; Neuropeptides ; Avian brain ; Domestic mallard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of immunoreactive thyrotropin-releasing hormone (TRH) in the central nervous system of the domestic mallard was studied by means of the peroxidase-antiperoxidase technique. After colchicine pretreatment, the highest number of TRH-immunoreactive perikarya was found in the parvocellular subdivision of the paraventricular nucleus and in the preoptic region; a smaller number of immunostained perikarya was observed in the lateral hypothalamic area and in the posterior medial hypothalamic nucleus. TRH-immunoreactive nerve fibers were detected throughout the hypothalamus, forming a dense network in the periventricular area, paraventricular nucleus, preoptic-suprachiasmatic region, and baso-lateral hypothalamic area. TRH-containing nerve fibers and terminals occurred in the organon vasculosum of the lamina terminalis and in the external zone of the median eminence in juxtaposition with hypophyseal portal vessels. Scattered fibers were also seen in the internal zone of the median eminence and in the rostral portion of the neural lobe. Numerous TRH-immunoreactive fibers were detected in extra-hypothalamic brain regions: the highest number of immunoreactive nerve fibers was found in the lateral septum, nucleus accumbens, olfactory tubercle, and parolfactory lobe. Moderate numbers of fibers were located in the basal forebrain, dorsomedial thalamic nuclei, hippocampus, interpeduncular nucleus, and the central gray of the mesencephalon. The present findings suggest that TRH may be involved in hypophysiotropic regulatory mechanisms and, in addition, may also act as neuromodulator or neurotransmitter in other regions of the avian brain.
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    URL: http://dx.doi.org/10.1007/BF00215853
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    Abundant GABAergic innervation of rat posterior pituitary revealed by inhibition of GABA-transaminase (1988)
    Springer
    Cell & tissue research 251 (1988), S. 59-64 
    Details
    ISSN: 1432-0878
    Keywords: Posterior pituitary ; Immunocytochemistry ; Anti-GABA ; GABA-transaminase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antibody against gamma-aminobutyric acid (GABA) was used to identify GABAergic elements immunocytochemically in the rat posterior pituitary. In order to increase the intracellular concentration of GABA, rats were treated with the GABA-transaminase inhibitor gamma-vinyl-GABA (GVG). Light-microscopic observations of Vibratome and semithin sections revealed the presence of numerous immunoreactive nerve fibers throughout the neural lobe; the mean number and length of these fibers increased by 90% after GVG treatment. Electron microscopy demonstrated the immunostained axons to be of small diameter. The reaction product was confined to small vesicles. No immunostaining occurred in pituicytes. The richness of the GABAergic innervation of the neural lobe contrasts with previous reports using antibodies against glutamate decarboxylase and supports the idea that GABA participates in the presynaptic control of neurosecretion.
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    URL: http://dx.doi.org/10.1007/BF00215447
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    Localization of avian LHRH-immunoreactive neurons in the hypothalamus of the domestic fowl, Gallus domesticus, and the Japanese quail, Coturnix coturnix (1988)
    Springer
    Cell & tissue research 251 (1988), S. 51-58 
    Details
    ISSN: 1432-0878
    Keywords: Hypothalamus ; LHRH-containing neurons ; Chicken LHRH ; Chicken GnRH-II ; Immunocytochemistry ; Domestic fowl ; Japanese quail
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of LHRH-containing perikarya and nerve fibers in the hypothalami of the domestic fowl and Japanese quail was investigated by means of the specific immunoperoxidase ABC method, using antisera against chicken LHRH-I ([Gln8]-LHRH), chicken GnRH-II ([His5-Trp7-Tyr8]-LHRH [2–10]) and mammalian LHRH ([Arg8]-LHRH). Chicken LHRH-I-immunoreactive perikarya were sparsely scattered in the nucleus preopticus periventricularis (POP), nucleus filiformis (FIL) and nucleus septalis medialis (SM), and in bilateral bands extending from these nuclei into the septal area in both species. A few reactive perikarya were also observed in the nucleus accumbens (Ac) and lobus parolfactorius (LPO). Numerous cLHRH-I-immunoreactive fibers were widely scattered in the preoptic, septal and tuberal areas, and were densely concentrated in the external layer of the median eminence and in organum vasculosum of the lamina terminalis (OVLT) in both species. Anti-mammalian LHRH serum cross-reacted weakly with perikarya and fibers immunoreactive to anti-cLHRH-I serum in normal chicken and quail. Anti-cGnRH-II[2–10] serum immunoreacted with magnocellular neurons distributed in the rostral end of the mesencephalon along the midline close to the nervus oculomotorius (N III). These perikarya were apparently different from cLHRH-I immunoreactive neurons. No immunoreactive cells and fibers against anti-cGnRH-II[2–10] were observed in the hypothalamus and median eminence of the chicken or quail. Anti-cGnRH-II[2–10] bound specifically with cGnRH-II. The morphological evidence suggests that cGnRH-II may not be secreted into the portal circulation to act as hypothalamic hormone.
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    URL: http://dx.doi.org/10.1007/BF00215446
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    Substance P-like immunoreactivity in the stomatogastric nervous systems of the crab Cancer borealis and the lobsters Panulirus interruptus and Homarus americanus (1988)
    Springer
    Cell & tissue research 252 (1988), S. 515-522 
    Details
    ISSN: 1432-0878
    Keywords: Substance P-related peptides ; Immunocytochemistry ; Invertebrate ganglia ; Peptidergic neurons ; Stomatogastric nervous system ; Cancer borealis, Panulirus interruptus, Homarus americanus (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of substance P-like immunoreactivity in the stomatogastric nervous systems of three decapod crustacean species, Cancer borealis, Homarus americanus, and Panulirus interruptus, was studied. The stomatogastric ganglion showed dense staining in the neuropil, but none in the somata. A single neuron stained in the esophageal ganglion. Lucifer yellow backfills and intracellular injections followed by incubation with the substance P antibody showed that the axons of this neuron project into the inferior esophageal nerves towards the paired commissural ganglia. The commissural ganglia showed a pronounced projection from a large bundle of fibers in the anterior medial portion of the circumesophageal connective. Additionally, less dense neuropil and stained somata were seen in the commissural ganglia. Staining was completely blocked by preabsorption with authentic substance P, physalaemin, eledoisin, and substance K. These data suggest that in the nervous system of crustacean species a molecule with C-terminal homology to substance P and other tachykinins is released as a neuroregulator in the stomatogastric ganglion.
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    URL: http://dx.doi.org/10.1007/BF00216638
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    Diversity of prolactin systems in the insect Leucophaea maderae: Use of antiserum polyclonality for immunocytochemical detection of neuropeptide heterogeneity (1988)
    Springer
    Cell & tissue research 252 (1988), S. 557-563 
    Details
    ISSN: 1432-0878
    Keywords: Prolactin-like neuropeptides ; Immunocytochemistry ; Brain ; Neuroendocrine structures ; Insects ; Leucophaea maderae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of prolactin-like neuropeptides was demonstrated immunocytochemically in the brain and affiliated neuroendocrine structures of the insect Leucophaea maderae. Use of the unlabelled peroxidase-antiperoxidase method of Sternberger revealed a rather widespread and differential distribution of reaction products resembling human (hPRL) and ovine (oPRL) prolactin. Tests with antirat PRL antibody were negative. The specificity of the antibodies used was established by liquid-phase absorptions and confirmed in tissue control systems. In L. maderae, anti-oPRL identifies part of an oPRL-like molecule different from human and rat PRL. Anti-hPRL reveals part of a human and ovine PRL-like molecule different from rat prolactin. These results indicate the occurrence, in the nervous tissue of one insect species, of at least two types of prolactin-like molecules.
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    URL: http://dx.doi.org/10.1007/BF00216642
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    Retention of carboxypropeptides in type-II collagen fibrils in chick embryo chondrocyte cultures (1988)
    Springer
    Cell & tissue research 252 (1988), S. 619-624 
    Details
    ISSN: 1432-0878
    Keywords: Collagen ; Carboxypropeptide ; Chondrocyte ; Tissue culture ; Immunocytochemistry ; Immunofluorescence ; Chick embryo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antibody reacting with the C-propeptide of chick type-II procollagen was used in an attempt to localize this terminal extension of the procollagen molecule (by immunogold labelling) during early collagen fibrillogenesis in chondrocyte cultures. After 2 days in culture the chondrocytes were surrounded by pericellular type-II collagen, as demonstrated by an indirect immunofluorescence labelling technique. An electron microscopy study of these cultures showed that the collagen fibrils were thin (∼ 15 nm diameter), with a poorly visible cross striation, sometimes enhanced by slight thickenings. The antibody against the C-propeptide of type-II procollagen labelled most of the collagen fibrils, according to a very regular pattern constituting a 60 nm periodicity. After 3 days the label was still present on the pericellular collagen fibrils but disappeared from the collagen fibrils of the extracellular matrix. Our results indicate that the C-propeptide of type-II procollagen is retained in the newly formed fibrils.
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    URL: http://dx.doi.org/10.1007/BF00216649
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    Immunocytochemical localisation of some lysosomal hydrolases, their presence in luminal fluid and their directional secretion by human epididymal cells in culture (1995)
    Springer
    Cell & tissue research 280 (1995), S. 415-425 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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    URL: http://dx.doi.org/10.1007/BF00307815
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