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  • Rat  (190)
  • Nitrogen fixation  (118)
  • Springer  (308)
  • American Association for the Advancement of Science
  • 2010-2014  (2)
  • 1980-1984  (306)
  • 1925-1929
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1004-1006 
    ISSN: 1420-9071
    Keywords: Rat ; adrenocortical responsiveness ; ACTH ; plasma ; corticosterone ; plasma ; corticotropin releasing factor (CRF)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the course of studying the plasma adrenocorticotropic hormone (ACTH) and corticosterone responses to synthetic corticotropin releasing factor (CRF), we noted some disparity in the responses. A higher dose (20 μg compared with 5 μg per rat i.a.) produced an equal plasma ACTH but greater plasma corticosterone response in adult male rats. Thus, we examined the possibility that CRF increases adrenocortical responsiveness to ACTH. CRF significantly (p〈0.0005) increased the plasma corticosterone response to ACTH in rats pretreated with dexamethasone. Thus, synthetic CRF increases corticosterone secretion in rats not only by stimulating ACTH secretion, but also by increasing the adrenocortical responsiveness to ACTH.
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  • 2
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    Cellular and molecular life sciences 40 (1984), S. 974-975 
    ISSN: 1420-9071
    Keywords: Rat ; prostaglandins ; gastric lesion ; intragastric distension model ; stress model ; indomethacin ; somatostatin preventive effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The inhibition of endogenous prostaglandin synthesis by indomethacin treatment blocks the somatostatin preventive effect on the gastric lesions induced in a stress model and has no preventive effect on an intragastric distension model.
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  • 3
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    Journal of molecular evolution 21 (1984), S. 19-32 
    ISSN: 1432-1432
    Keywords: Leghemoglobin ; Gene duplication ; Gene linkage ; Concerted evolution ; Nitrogen fixation ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed the sequences of soybean leghemoglobin genes as an initial step toward understanding their mode of evolution. Alignment of the sequences of plant globin genes with those of animals reveals that (i) based on the proportion of nucleotide substitutions that have occurred at the first, second, and third codon positions, the time of divergence of plant and animal globin gene families appears to be extremely remote (between 900 million and 1.4 billion years ago, if one assumes constancy of evolutionary rate in both the plant and animal lineages) and (ii) in addition to the normal regulatory sequences on the 5′ end, an approximately 30-base-pair sequence, specific to globin genes, that surrounds the cap site is conserved between the plant and animal globin genes. Comparison of the leghemoglobin sequences with one another shows that (i) the relative amount of sequence divergence in various coding and noncoding regions is roughly similar to that found for animal globin genes and (ii) as in animal globin genes, the positions of insertions and deletions in the intervening sequences often coincide with the locations of direct repeats. Thus, the mode of evolution of the plant globin genes appears to resemble, in many ways, that of their animal counterparts. We contrast the overall intergenic organization of the plant globin genes with that of animal genes, and discuss the possibility of the concerted evolution of the leghemoglobin genes.
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  • 4
    ISSN: 1420-9071
    Keywords: Rat ; cerebrospinal fluid, human ; analgesia ; naloxone ; pain indifference, congenital ; opiates, endogenous
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary CSF from a patient with congential indifference to pain was found to produce analgesia in the rat following intracerebroventricular injections. The analgesic effect was attenuated by pretreatment with naloxone suggesting the involvement of hyperactive endogenous opiate mechanisms in this patient.
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  • 5
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    Cellular and molecular life sciences 40 (1984), S. 1368-1369 
    ISSN: 1420-9071
    Keywords: Rat ; kidney ; hypertensive ; prostaglandin dehydrogenase ; hexokinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 15-Hydroxyprostaglandin dehydrogenase (PGDH) surged in hypertensive (SHR) and normotensive (WKY) rat kidney at 8 days of age, is greatest in SHR. Hexokinase fell in SHR at 17 days of age, but thereafter was similar to WKY. This suggests multisystem enzymatic abnormalities in SHR kidney during development of hypertension.
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  • 6
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    Cellular and molecular life sciences 40 (1984), S. 1008-1010 
    ISSN: 1420-9071
    Keywords: Rat ; ethanol preference ; acetaldehyde self-administration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Peripherally self-injected acetaldehyde in interaction with environmental and nutritional variables significantly enhances alcohol drinking in rats and suggests an involvement of acetaldehyde in voluntary alcohol intake.
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  • 7
    ISSN: 1432-0827
    Keywords: Rat ; Calvarium ; Electron microscopy ; Preosteoclasts ; Osteoclasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary This is a study of the fine structure of cells of the 20-day fetal rat calvarium. Special attention is given to identifying and characterizing preosteoclasts. These cells are relatively common and located largely, but not exclusively, at the endocranial bone surface. The preosteoclasts are characterized by abundant mitochondria, an incomplete perinuclear Golgi apparatus, and variable-shaped dense granules. The dense granules are unique in appearance in that they contain an internal dense matrix surrounded by a clear halo. Most granules are circular in shape but some are elongate or tubular in form. Granules with identical appearance are observed in osteoclasts. The preosteoclasts are mononucleate, or occasionally binucleate. It is suggested that because preosteoclasts are morphologically distinctive and relatively abundant, it should be feasible to separate these cells from a heterogeneous cell isolate.
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  • 8
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    Calcified tissue international 34 (1982), S. 211-213 
    ISSN: 1432-0827
    Keywords: Rat ; incisor ; ameloblasts ; enamel ; 45Ca autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Rats were injected with45Ca and horseradish peroxidase to determine the patterns of45Ca incorporation into incisor enamel and the morphological types of the overlying maturation ameloblasts.45Ca autoradiography showed no differences in the patterns of incorporation into enamel between routinely embedded and freeze-dried specimens. Enamel overlaid by ruffle-ended ameloblasts was much more heavily labeled while that overlaid by smooth-ended ameloblasts showed only moderate labeling. The observations lend further support to the hypothesis that the ruffle-ended cells are very active in mineralizing enamel and that the smooth-ended cells are in a passive, restorative phase.
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  • 9
    ISSN: 1432-0827
    Keywords: Connective Tissue ; Vitamin D deficiency ; Hydroxyproline excretion ; Glycosaminoglycan excretion ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The urinary excretion of two connective tissue metabolites was studied in both control and vitamin D deficient rats. Hydroxyproline (HyPRO) excretion was determined after 2, 13 and 22 months (experiment I). It decreased with aging in animals receiving the control diet. On the contrary, this excretion increased as a function of age in vitamin D deficient animals. At the age of 22 months, HyPRO excretion was respectively 31 and 1708 µg a day in control and deficient animals. HyPRO and glycosaminoglycans (GAG) excretion was measured on a group of both control and vitamin D deficient rats at the age of 21 months (experiment II). These results confirm the high excretion of HyPRO in deficient animals. On the contrary, the GAG excretion was higher in control animals than in deficient ones, the mean excretion being respectively 412 and 234 µg a day.
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  • 10
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    Calcified tissue international 32 (1980), S. 63-68 
    ISSN: 1432-0827
    Keywords: Rat ; Bone formation ; Fluorochrome ; Microphotometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A method for quantitative studies of the formation rate of bone has been developed. After vital staining with calcein, the fluorescence of a bone section was measured with a microphotometer controlled by a mini computer. After staining the bone structure with alizarin red S in a second step, the section was measured in transmitted light. The two data sets were combined and the shortest distances between the bone surface and the fluorescence lines were computed. With this information the distance distribution and the bone area between the label and the surface could be calculated in two different ways: with the single labeling and the continuous labeling techniques. The advantages and disadvantages of the two methods are discussed and compared with those of other techniques.
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  • 11
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    Calcified tissue international 32 (1980), S. 113-122 
    ISSN: 1432-0827
    Keywords: Rat ; Bone ; Metaphysis ; Quantitative ; Aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of this work was to analyze the proximal tibial metaphysis of the 170 g rat in a quantitative histologic fashion which would allow some relation to tissue age to be established. Stained 3 µm thick tissue sections were analyzed with the aid of a Merz grid on an eyepiece reticule and a light microscope. Tissue mass and cell distribution were studied in all areas. The rate of change in tissue mass during aging of the metaphysis was calculated. Two regions of the metaphysis were identified. One, corresponding to the primary spongiosa, less than 4.45 days of age, is a region of high turnover of hard tissue and high numbers of osteoblasts and osteoprogenitor cells. The other, corresponding to the secondary spongiosa, is a region of relatively low net tissue turnover and low numbers of osteoblasts and osteoprogenitor cells. Osteoclasts were found relatively more uniformly distributed through the metaphysis than were osteoblasts and osteoprogenitor cells. The rate of bone formation in the primary spongiosa is 50 times that found in the Haversian bone of the rib of 5-year-old humans and about 500 times that found at the cortical-endosteal surface of ribs of 5-year-old humans. It is argued that both cell distribution and tissue distribution in the metaphysis support the concept that osteoblasts and osteoclasts, rather than osteocytes, are responsible for the maturation of the metaphysis. The inhomogeneous distribution of both cells and tissue in the metaphysis has definite meaning for the interpretation of findings concerning the incorporation of radionuclides into the skeleton.
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  • 12
    ISSN: 1432-0827
    Keywords: Rat ; Hypophysectomy ; Dietary phosphorus deprivation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The demands of growth are known to exacerbate the effect of phosphorus deprivation (PD). We examined whether changes associated with PD could be prevented in young rats in which growth and growth hormone (GH) were eliminated by hypophysectomy (HPX) and whether PD in normal intact rats (INT) was associated with increased secretion of GH. INT or thyroxine- and ACTH-replaced HPX rats were fed one of the three diets: 0.31% P (NP); 0.027% P (LP), and 0.31% P, pair-fed with LP-mates (NP-PF). The results indicate that HPX did not qualitatively alter several physiologic responses to PD: (a) serum and urinary phosphorus (P) decreased and urinary calcium (Ca) increased; (b) net intestinal Ca retention fell and duodenal sac uptake of45Ca rose; and (c) external P balance was restored and duodenal sac uptake of32P-phosphate increased. Only the hypercalcemia seen in INT, LP rats was prevented by HPX. In INT rats serum immunoassayable GH levels, measured in single samples, were not different between different dietary groups while pituitary bioassayable GH was reduced in both LP and NP-PF rats when compared to the NP rats. Thus, except for hypercalcemia, the physiologic responses associated with PD are not prevented by the elimination of growth and GH, and the development of these responses in INT rats was not associated with a consistent or specific alteration in GH secretion.
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  • 13
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    Calcified tissue international 33 (1981), S. 223-231 
    ISSN: 1432-0827
    Keywords: Protein deficiency ; Bone formation ; Skeletal development ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Protein deficiency was produced by freely feeding young rats a 1% lactalbumin diet for 12 weeks in order to study the effects of protein-calorie malnutrition on skeletal development. During the experiment the food and caloric intake and weight of the experimental animals decreased, while those parameters of the control animals progressively increased. However, when gross caloric intake was expressed as a function of the metabolic size of the animal, the caloric consumption was similar for both groups of animals. The protein-deficient animals exhibited micro-radiographic and histological features of an abnormal pattern of endochondral bone formation. Appositional bone growth, as determined by the daily appositional rate and the percentage of endosteal surfaces undergoing active bone formation, was significantly decreased in these animals, as was the percentage of periosteal surfaces exhibiting resorption. Both chemical analyses of the whole bone and electron probe microanalysis in the specific area of actively calcifying bone revealed no significant differences between the mineral content of control and protein-deficient animals. This study distinguishes the effects of protein deficiency from that of combined protein-calorie deprivation and demonstrates that the abnormal skeletal development observed was the result of a decrease in the quantity of bone formed rather than an altered mineral content.
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  • 14
    ISSN: 1432-0827
    Keywords: Rat ; Incisor ; Amelogenesis ; Acid phosphatase ; Ferritin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Acid phosphatase was localized in rat incisor ameloblasts without prior decalcification. Whenβ-glycerophosphate was used as the substrate, an intense reaction was observed in the supranuclear region of the secretory ameloblasts. But the reaction was dramatically reduced at the transitional stage and was very weak in the maturation ameloblasts. Whenp-nitrophenylphosphate was the substrate, the reaction product was consistently seen in the Golgi cisternae and the vesicular components of the ameloblasts at all stages of enamel development. These observations suggest that there are two acid phosphatases in ameloblasts. One is in the secretory ameloblasts and the other in the transition and maturation ameloblasts. X-ray micro-analyses for Fe and Pb showed that Fe and acid phosphatase were in the ferritin-containing vesicles at the later stage of enamel maturation. This evidence suggests that ferritin is digested in these vesicles for the release of the Fe pigment to the enamel. An increase in the number of intercellular bridges between ameloblasts was correlated with the dramatic decrease in height of ameloblasts at the pigment release stage. The ameloblast membranes were acid phosphatase positive at the intercellular bridges whenp-nitrophenylphosphate was the substrate. This activity may be involved in the reduction in the surface area of the ameloblast membranes.
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  • 15
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    Calcified tissue international 34 (1982), S. 376-381 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Bone ; Actin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Preliminary indications of the occurrence of actin and myosin in crude matrix vesicle preparations have been reported previously. In the present study extracellular matrix vesicles from rat alveolar bone were isolated. They were further purified by a sucrose density gradient. SDS-polyacrylamide gel electrophoresis of the purified vesicles revealed the presence of a polypeptide with a molecular weight of 43 K daltons and with electrophoretic mobility identical to that of blood platelet actin. The limited proteolysis of both 43 K dalton vesicular polypeptide and actin byStaphylococcus aureus-V8-protease revealed three fragments with identical electrophoretic mobility. In addition, the vesicular preparations inhibited the activity of DNase I, a property typical of actin monomers. Filamentous material extracted from matrix vesicles showed ultrastructural features of F-actin. Reaction of this material with heavy meromyosin resulted in arrowhead formation, which is characteristic of acto-heavy meromyosin. The occurrence of actin in extracellular matrix vesicles may account for their budding from the osteoblastic plasma membrane, their possible motility in the matrix, and maintenance of the spherical shape.
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  • 16
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    Calcified tissue international 35 (1983), S. 107-110 
    ISSN: 1432-0827
    Keywords: Calcium ; Glucocorticoid ; Vitamin D ; Osteoporosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Eighty-eight adult male Sprague-Dawley rats were given a diet with either (a) 0.5% Ca and 0.6% P or (b) 0.01% Ca and 0.6% P. Osteopenia was created by adding prednisolone to the diet. The prophylactic effect of oral 1,25(OH)2D3 on the osteopenia was studied. It was found that prednisolone osteopenia in the rat was associated with defective Ca absorption. By giving an oral dose of 1,25(OH)2D3, it was possible to maintain normal Ca absorption during prednisolone treatment and to prevent the bone loss. No significant hypercalcemia or any kidney calcifications were seen. These results are in contrast to earlier findings, in which subcutaneous administration of 1,25(OH)2D3 failed to prevent prednisolone osteopenia because of its tendency to increase bone resorption.
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  • 17
    ISSN: 1432-2048
    Keywords: Alnus ; Ammonium ; Carbon translocation ; Endophyte damage ; Nitrogen fixation ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cloned plants of Alnus incana (L.) Moench were inoculated and grown without combined nitrogen for seven weeks. The effects of ammonium on the function and structure of the root nodules were studied by adding 20 mM NH4Cl (20 mM KCl=control) for four days. Nitrogenase activity decreased to ca. 50% after one day and to less than 10% after two days in ammonium treated plants, but was unaffected in control plants. The results were similar at photon flux densities of 200 and 50 μmol m-2 s-1. At the higher light level the effect was concentration dependent between 2 and 20 mM NH4Cl. The recovery was slow, and more than 11 d were needed for plants treated with 20 mM ammonium to reach initial activity. The distribution of 14C to the root nodules after assimilation of 14CO2 by the plants was not changed by the ammonium treatment. Microscopical studies of root nodules showed high frequencies of endophyte vesicles being visually damaged in nodules from ammonium-treated plants, but not in nodules from control plants. When nitrogenase activity was restored, visually damaged vesicles were again few, whereas young developing vesicles were numerous. The slow recovery, the 14C-translocation pattern, and the structural changes of the endophyte indicate a more complex mechanism of ammonium influence than simply a short-term reduction in supply of carbon compounds to the nodules.
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  • 18
    ISSN: 1432-2048
    Keywords: Ammonia ; Nitrogen fixation ; Nodule ; Senescence (root nodules) ; Ureide ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract During early development (up to 18 d after sowing) of nodules of an “effective” cowpea symbiosis (Vigna unguiculata (L.) Walp cv. Vita 3: Rhizobium strain CB756), rapidly increasing nitrogenase (EC 1.7.99.2) activity and leghaemoglobin content were accompanied by rapid increases in activities of glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 2.6.1.53), enzymes of denovo purine synthesis (forming inosine monophosphate) xanthine oxidoreductase (EC 1.2.3.2), urate oxidase (EC 1.7.3.3), phosphoenolpyruvate carboxylase (EC 4.1.1.31) and led to increased export of ureides (allantoin and allantoic acid) to the shoot of the host plant in the xylem. Culturing plants with the nodulated root systems maintained in the absence of N2 (in 80 Ar: 20 O2, v/v) had little effect on the rates of induction and increase in nitrogenase activity and leghaemoglobin content but, in the absence of N2 fixation and consequent ammonia production by bacteroids, there was no stimulation of activity of enzymes of ammonia assimilation or of the synthesis of purines or ureides. Addition of NO 3 - (0.1–0.2 mM) relieved host-plant nitrogen deficiency caused by the Ar: O2 treatment but failed to increase levels of enzymes of N metabolism in either the bacteroid or the plant-cell fractions of the nodule. Premature senescence in Ar: O2-grown nodules occurred at 18–20 d after sowing, and resulted in reduced levels of nitrogenase activity and leghaemoglobin but increased the activity of hydroxybutyrate oxidoreductase (EC 1.1.1.30).
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  • 19
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Glycine ; Nitrogen fixation ; Proplastid ; Purine synthesis ; Root nodule ; Ureide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Subcellular organelle fractionation of nitrogen-fixing nodules of soybean (Glycine max (L.) Merr.) indicates that a number of enzymes involved in the assimilation of ammonia into amino acids and purines are located in the proplastids. These include asparagine synthetase (EC 6.3.1.1), phosphoribosyl amidotransferase (EC 2.4.2.14), phosphoglycerate dehydrogenase (EC 1.1.1.95), serine hydroxymethylase (EC 2.1.2.1), and methylene-tetrahydrofolate dehydrogenase (EC 1.5.1.5). Of the two isoenzymes of asparate aminotransferase (EC 2.6.1.1) in the nodule, only one was located in the proplastid fraction. Both glutamate synthase (EC 1.4.1.14) and triosephosphate isomerase (EC 5.3.1.1) were associated at least in part with the proplastids. Glutamine synthetase (EC 6.3.1.2) and xanthine dehydrogenase (EC 1.2.1.37) were found in significant quantities only in the soluble fraction. Phosphoribosylpyrophosphate synthetase (EC 2.7.6.1) was found mostly in the soluble fraction, although small amounts of it were detected in other organelle fractions. These results together with recent organelle fractionation and electron microscopic studies form the basis for a model of the subcellular distribution of ammonium assimilation, amide synthesis and uredie biogenesis in the nodule.
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  • 20
    ISSN: 1432-2048
    Keywords: Allantoin ; Amino acids ; Bleeding sap ; Nitrogen fixation ; Phaseolus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dwarf french beans, Phaseolus vulgaris L., were grown with or without inoculation with rhizobia (strain 3644), and with or without a combined nitrogen source (nitrate or ammonium ions). The distribution of radioactivity into products of dark 14CO2 assimilation was studied in roots or nodules from these plants. A detailed study was also made of the distribution and rates of excretion of nitrogen in xylem bleeding sap in 28 day old plants grown on the various sources of nitrogen. Whereas detached nodules accumulated radioactive glycine, serine and glutamate when incubated with 14CO2, bleeding sap from plants root fed 14CO2 contained low levels of radioactivity in these compounds but higher levels in allantoin. Chemical analysis showed allantoin to be the major compound transported in the xylem of nodulated plants, whether or not they were fed on combined nitrogen. In contrast uninoculated plants accumulated mainly amino acids in the bleeding sap, the amount and chemical composition of which depended on the combined nitrogen source.
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  • 21
    ISSN: 1432-2048
    Keywords: Glyoxylate ; Isonicotinic acid hydracide ; Medicago ; Nitrogen fixation ; Photorespiration ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation (measured as acetylene reduction) by whole nodulated alfalfa plants was stimulated when the plants were treated with isonicotinic acid hydracide (INH) and glyoxylate, both inhibitors of the glycolate pathway of carbohydrate metabolism, at concentrations of 300 and 100 mM, respectively. Reducing energetic loses caused by photorespiration results in an increase in the symbiotic nitrogen fixation.
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  • 22
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    Planta 152 (1981), S. 544-552 
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Lichens ; Nitrogen fixation ; Peltigera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Nostoc in the cephalodia of the lichen Peltigera aphthosa Willd. fixed 15N2 and the bulk of the nitrogen fixed was continuously transferred from it to its eukaryotic partners (a fungus and a green alga, Coccomyxa sp.). Kinetic studies carried out over the first 30 min, after exposure of isolated cephalodia to 15N2, showed that highest initial 15N2-labelling was into NH 4 + . After 12 min little further increase in the NH 4 + label occurred while that in the amide group of glutamine and in glutamate continued to increase. The 15N-labelling of the amino group of glutamine and of aspartate increased more slowly, followed by an increase in the labelling of alanine. When total incorporation of 15N-label was calculated, the overall pattern was found to be rather similar except that, throughout the experiment, the total 15N incorporated into glutamate was about six times greater than that into the amide group of glutamine. Pulse chase experiments, in which 14N2 was added to cephalodia previously exposed to 15N2, showed that the NH 4 + pool rapidly became depleted of 15N-label, followed by decreases in the labelling of glutamate, the amide group of glutamine and aspartate. The 15N-labelling of alanine, however, continued to increase for a period. When isolated cephalodia were treated with L-methionine-SR-sulphoximine, an inhibitor of glutamine synthetase (EC 6.3.1.2), and azaserine, an inhibitor of glutamate synthase (EC 2.6.1.53), there was no detectable labelling in glutamine although the 15N-labelling of glutamate increased unimpaired. On treating the cephalodia with amino-oxyacetate, an inhibitor of aminotransferase activity, the alanine pool decreased. Evidence was obtained that glutamine synthetase and glutamate synthase were located in the Nostoc, and that glutamate dehydrogenase (EC 1.4.1.4) and various amino-transferases were located in the cephalodial fungus. Possible implications of these findings are discussed.
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  • 23
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    Environmental management 7 (1983), S. 177-187 
    ISSN: 1432-1009
    Keywords: Nitrogen ; Model ; Agriculture ; Mass balance ; Ground-water ; Denitrification ; Immobilization ; Dry deposition ; Nitrogen fixation ; Nitrate ; Florida
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract A detailed nitrogen budget was devised for agricultural activities in the Florida peninsula, based on routine data published by state agricultural agencies. The model demonstrates that important unmonitored fluxes of nitrogen can often be calculated by mass balance on individual model compartments, and that the reasonability of poorly quantified fluxes can be assessed. The results of such models can be very useful in designing and assessing the results of field experiments and in prioritizing environmental monitoring programs.
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  • 24
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Regulation ; Guanosine 5′-diphosphate 3′-diphosphate (ppGpp)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Addition of ammonium to N2 fixing cultures of Azotobacter vinelandii, Klebsiella pneumoniae and Clostridium pasteurianum rapidly reduced the intracellular levels of guanosine 5′-diphosphate 3′-diphosphate (ppGpp) by 70–90%. This change might reflect a regulatory role of ppGpp in nitrogen metabolism.
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  • 25
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    Archives of microbiology 136 (1983), S. 20-25 
    ISSN: 1432-072X
    Keywords: Hydrogen production ; Nitrogen fixation ; Hydrogen recycling ; Hydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutants of Rhodopseudomonas capsulata lacking uptake hydrogenase activity have been isolated among those unable to grow photoautotrophically. Studies with these mutants showed increases in nitrogenase mediated H2 production from all substrates tested. In addition, photosynthetic synthetic growth on N2 with malate as carbon source was not affeced by the block in H2 uptake even under low light. Under these growth conditions hydrogen was observed to accumulate in mutant but not in wild-type cultures. This finding suggested that H2 was evolved by nitrogenase during N2 fixation by this photosynthetic bacterium and was efficiently recycled in the wild type.
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  • 26
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    Archives of microbiology 135 (1983), S. 287-292 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Respiration ; Nitrogen fixation ; Heterocysts ; K m for O2 ; Anabaena variabilis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Simultaneous measurements of acetylene reduction by Anabaena variabilis and the concentration of dissolved oxygen in the suspension were made using a specially designed vessel which allowed measurements under steady-state conditions. The rate of acetylene reduction in the dark increased with increasing oxygen concentrations until a maximum value was reached at 300 μM O2 (corresponding to 30% O2 in the gas phase at 35°C). This presumably results from a requirement for energy provided by respiration. Measurements of the dependence of respiration rate on dissolved oxygen concentration were made under comparable conditions using an open system to allow conditions close to steady-state to be obtained. The respiration rate of diazotrophically grown Anabaena variabilis had a dependence on oxygen concentration corresponding to the sum of two activities. These had K m values of 1.0 μM and 69 μM and values of V max of similar magnitude. Only the high affinity activity was observed in nitrate-grown cyanobacteria lacking heterocysts, and this presumably represent activity in the vegetative cells. The oxygen concentration dependence of the low affinity activity resembled that for the stimulation of acetylene reduction. We interpret this as the result of oxygen uptake by the heterocysts. The results are consistent with the idea that in intact filaments of cyanobacteria O2 enters heterocysts much more slowly than it enters the vegetative cells.
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  • 27
    ISSN: 1432-072X
    Keywords: Ammonium export ; Ammonium assimilation ; Glutamine synthetase ; Nitrogen fixation ; Rhizobium sp. 32H1
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    Notes: Abstract The relationship between ammonium assimilation and ammonium export has been studied in free-living, N2-fixing Rhizobium sp. 32H1. After 55 to 67 h of microaerobic growth under a gas phase of 0.2% O2 – 1.0% CO2 – 98.8% Ar high levels of nitrogenase were observed concomitant with a slightly adenylylated glutamine synthetase (GSI) and some glutamine synthetase (GSII) activity. However, after growth of 89 h, or longer, GSI became adenylylated and the level of GSII had decreased. When the gas phase was shifted to 0.2% O2 – 1.0% CO2 – 98.8% N2, a lag was observed before ammonium export could be detected in the 55 to 67 h cultures. No lag in ammonium export was observed in the cultures previously grown for 89 h. The onset of ammonium export in the 55 to 67 h cultures was found to correlate with the adenylylation state of GSI. There appeared to be no correlation between the level of GSII and the export of ammonium. Neither an increase in the adenylylation level of GSI nor ammonium export was observed when the 55 to 67 h cultures were maintained under the Ar gas mixture.
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  • 28
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    Archives of microbiology 127 (1980), S. 163-165 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Arthrobacter ; Corynebacterium ; Anabaena azollae
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    Topics: Biology
    Notes: Abstract Coryneform bacteria were found associated with the nitrogen fixing blue-green alga, Anabaena azollae in the leaf cavity of Azolla caroliniana. Plate counts indicated ca. 7,400±1,900 bacterial cells per mature leaf cavity or approximately 1 bacterial cell for every algal cell. No other type of bacterium was found in these cavities.
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  • 29
    ISSN: 1432-072X
    Keywords: Fluorescent antibody staining ; Nitrogen fixation ; Symbiosis ; Anabaena azollae ; Azolla caroliniana
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    Notes: Abstract Fluorescent antibody staining indicated differences in surface antigenicity in Anabaena azollae cells fresh from the leaf cavities of the fern, Azolla caroliniana, and algae which were isolated and subcultured from this fern. Such results suggest that either changes in antigenicity occur in this phycobiont during culturing or that isolation selects for an antigenically different mutant strain capable of in vitro growth.
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  • 30
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    Archives of microbiology 129 (1981), S. 238-239 
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    Keywords: Rhizobium ; Disaccharide ; Bacteroid ; Transport ; Nitrogen fixation
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    Topics: Biology
    Notes: Abstract Slow growing strains of rhizobia appear to lack both uptake systems and catabolic enzymes for disaccharides. In the fast-growing strains of rhizobia there are uptake mechanisms and catabolic enzymes for disaccharide metabolism. In Rhizobium leguminosarum WU 163 and WU235 and R. trifolii WU290, sucrose and maltose uptake appears to be constitutive whereas in R. meliloti WU60 and in cowpea Rhizobium NGR234 uptake of these disaccharides is inducible. There is evidence that there are at least two distinct disaccharide uptake systems in fast-growing rhizobia, one transporting sucrose, maltose and trehalose and the other, lactose. Disaccharide uptake is via an active process since uptake is inhibited by azide, dinitrophenol and carbonyl cyanide m-chlorophenylhydrazone but not by arsenate. Bacteroids of R. leguminosarum WU235 and R. lupini WU8 are unable to accumulate disaccharides.
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  • 31
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    Archives of microbiology 130 (1981), S. 38-43 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Gene expression ; Regulation ; Messenger RNA ; Transcription ; Klebsiella pneumoniae
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    Notes: Abstract Nitrogenase messenger RNA synthesis in Klebsiella pneumoniae was determined by labelling cells with (3H)uracil and isolating total RnA, which was then hybridized to filterbound recombinant plasmid pSA30 DNA carrying the nitrogenase structural genes nifH, D, and K. Derepression of nitrogenase mRNA starts 1.5 h before the onset of nitrogenase activity (as measured by acetylene reduction). Exposure of nif-derepressed cultures to either NH 4 + , air, or high temperatures (39° C) results in a rapid decrease of the synthesis rates both of nitrogenase mRNA and nitrogenase polypeptides. Nitrogenase mRNA is remarkably stable. After blocking transcription with rifampicin, hybridizable and actively translatable nitrogenase mRNA survives with an average half-life of 18 min. Half-lives are considerably shorter when rifampicin-inhibited cultures are simultaneously shifted to conditions which are non-permissive for nitrogenase synthesis, pointing to some posttranscriptional influence on nitrogenase mRNA stability. In all experiments performed there was no evidence for uncoupling of nitrogenase mRNA synthesis from nitrogenase mRNA translation, indicating that nitrogenase synthesis is regulated solely by transcriptional control.
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  • 32
    ISSN: 1432-072X
    Keywords: Rhizobium trifolii ; Symbiosis ; Nodulation ; Nitrogen fixation ; Symbiotic genes ; Reiterated sequences ; Plasmid
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    Notes: Abstract A Rhizobium trifolii symbiotic plasmid specific gene library was constructed and the physical organisation of regions homologous to nifHDK, nifA and nod genes was determined. These symbiotic gene regions were localised to u 25 kb region on the sym-plasmid, pPN1. In addition four copies of a reiterated sequence were identified on this plasmid, with one copy adjacent to nifH. No rearrangement of these reiterated sequences was observed between R. trifolii bacterial and bacteroid DNA. Analysis of a deletion derivative of pPN1 showed that these sequences were spread over a 110 kb region to the left of nifA.
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  • 33
    ISSN: 1432-072X
    Keywords: Ammonia assimilation ; Lichen symbioses ; Nitrogen fixation ; 15N kinetics ; Peltigera canina
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    Notes: Abstract On following N2-incorporation and subsequent metabolism in the lichen Peltigera canina using 15N as tracer, it was found, over a 30 min period, that greatest initial labelling was into NH 4 + followed by glutamate and the amide-N of glutamine. Labelling of the amino-N of glutamine, aspartate and alanine increased slowly. Pulse-chase experiments using 15N confirmed this pattern. On inhibiting the GS-GOGAT pathway using l-methionine-dl-sulphoximine and azaserine, 15N enrichment of glutamate, alanine and aspartate continued although labelling of glutamine was undetectable. From this and enzymic data, NH 4 + assimilation in the P. canina thallus appears to proceed via GS-GOGAT in the cyanobacterium and via GDH in the fungus; aminotransferases were present in both partners. The cyanobacterium assimilated 44% of the 15N2 fixed; the remainder was liberated almost exclusively as NH 4 + and then assimilated by fungal GDH.
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  • 34
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    Archives of microbiology 135 (1983), S. 103-109 
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    Keywords: RNA polymerase ; Transcription ; Nitrogen fixation ; Symbiosis ; Rhizobium
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    Notes: Abstract DNA-dependend RNA polymerase (EC 2.7.7.6) from Rhizobium japonicum was purified. The subunit structure was found to be ββ′α2σ, with the following apparent molecular weights determined by electrophoresis: M r (β and β') 150,000 each, M r (σ) 96,000, M r (α) 40,000, M r (holoenzyme) 490,000, M r (core enzyme) 380,000. The recovery of σ was 28%. RNA polymerase from aerobically grown R. japonicum cells and from nitrogen-fixing cells have the same electrophoretic properties suggesting that no chemical modification of the enzyme occurs when cells undergo this metabolic differentiation. The enzyme is Mg2+-dependent, rifampicin-sensitive, and has optimal activity at alkaline pH (8–10) and at 35–40° C. It binds strongly to bacteriophage T7 promoters, weakly to antibiotic resistance genes, and not at all to cloned R. japonicum nif DNA. Preliminary in vitro transcription experiments, including nif DNA as template, revealed that additional factors may be required for selective transcription from promoters.
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  • 35
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    Archives of microbiology 136 (1983), S. 219-221 
    ISSN: 1432-072X
    Keywords: Xanthobacter ; Nitrogen fixation ; Oxygen sensitivity ; Nitrogen metabolism ; Glutamine synthetase ; Glutamate synthase
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    Notes: Abstract N2-fixation was investigated in the chemoautotrophic hydrogen bacterium Xanthobacter H4-14. N2-fixing batch cultures of this organism could only be grown at pO2 values of around 0.02 bar, and in continuous culture dissolved oxygen tensions above 16 μM were found to inhibit N2-fixation. Xanthobacter H4-14 utilized a variety of amino acids, nitrate and ammonia as nitrogen sources. Cell-free extracts from steady-state continuous cultures of ammonia grown, nitrate grown and N2-fixing Xanthobacter were assayed for the presence of ammonia assimilation enzymes. No alanine dehydrogenase or glutamate dehydrogenase activity was detected. Ammonia was assimilated exclusively via the glutamine synthetase/glutamate synthase pathway, irrespective of the extracellular concentration of ammonia.
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  • 36
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    Archives of microbiology 124 (1980), S. 161-167 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Photosynthesis ; Green alga ; Chl a and b ; DCMU ; Light and O2 dependency
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    Notes: Abstract A unicellular alga which can grow in the light without a combined nitrogen source was isolated from a hot spring. The cells were almost spherical, usually 5–10 μm in diameter. Absorption spectra of the watersoluble pigments and of the acetone-extracted ones revealed the existence of chlorophyll a and b and the absence of phycobilins. Thin sections examined by electron microscopy revealed an eukaryotic organization with features typical of the coccoid green algae (the Chlorococcales). Cells divided by internal cytokinesis and subsequent liberation of daughter cells from the parental wall, in a way similar to Chlorella. The alga reduced acetylene to ethylene and incorporated 15N2 into cell protoplasm when incubated in a low oxygen atmosphere. Nitrogenase activity was light-dependent, microaerophilic and thermophilic. Although the association of symbiotic nitrogen fixing prokaryotes with the cells may still be possible, any such organisms have not so far been detected.
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  • 37
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    Archives of microbiology 127 (1980), S. 115-118 
    ISSN: 1432-072X
    Keywords: Proteus mirabilis ; Serratia marcescens ; Erwinia herbicola ; Nitrogen fixation ; nif genes ; his genes ; Plasmids
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    Notes: Abstract Plasmid pRD1, an R plasmid of the P incompatibility group which carries his and nif genes from Klebsiella pneumoniae in addition to drug resistance markers derived from RP4, was transferred to His- mutants of Serratia marcescens, Erwinia herbicola and Proteus mirabilis. His+ transconjugants were obtained at low but different frequencies according to recipient genus. Transconjugants all acquired the drug resistance, and were Nif+ in S. marcescens and E. herbicola, having acetylene-reducing activities of the same order of magnitude as the parent K. pneumoniae and fixing 15N2. No evidence for nif expression in P. mirabilis transconjugants was obtained though the nif genes were present.
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  • 38
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    Archives of microbiology 132 (1982), S. 219-224 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Nitrogen fixation ; Nodules ; Soybean
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    Notes: Abstract Several mutants defective in nodulation were isolated from Rhizobium japonicum strains 3I1b110 and 61A 76. Mutants of class I do not form nodules after incubation with soybean [Glycine max (L.) Merrill] for 17 days, but will do so by 28 days. When host plants other than G. max are infected with several of these strains, there is no detectable difference in the time of nodulation or size of nodules as compared to the wild type. Two mutants of class I (i. e., SM1 and SM2) have been shown previously to be altered in the lipopolysaccharide portion of their cell wall. Mutants of class II are not slow to nodulate but form fewer nodules than the wild type on all the host plants tested. Mutants of class III are unable to form nodules. Some bacteriophage-resistant mutants, altered in cell surface structure, fall into this class. Two mutants of class III do not bind to soybean roots.
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  • 39
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    Archives of microbiology 133 (1982), S. 172-177 
    ISSN: 1432-072X
    Keywords: Beggiatoa ; Nitrogen fixation ; Acetylene reduction ; Nitrate assimilation ; Microaerobic ; Isolation of marine strains
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    Notes: Abstract Four newly isolated marine strains of Beggiatoa and five freshwater strains were tested for nitrogen fixation in slush agar medium. All strains reduced acetylene when grown microaerobically in media containing a reduced sulfur source and lacking added combined nitrogen. The addition of 2 mmol N, as nitrate or ammonium salts, completely inhibited this reduction. Although not optimized for temperature or cell density, acetylene reduction rates ranged from 3.2 to 12 nmol·mg prot-1 min-1. Two freshwater strains did not grow well or reduce acetylene in medium lacking combined nitrogen if sulfide was replaced by thiosulfate. Two other strains grew well in liquid media lacking both combined nitrogen and reduced sulfur compounds but only under lowered concentrations of air. All freshwater strains grew well in medium containing nitrate as the combined nitrogen source. Since they did not reduce acetylene under these conditions, we infer that they can assimilate nitrate.
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  • 40
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    Archives of microbiology 133 (1982), S. 312-317 
    ISSN: 1432-072X
    Keywords: Rhizobium japonicum ; Rhizobium leguminosarum ; Formate metabolism ; Formate dehydrogenase ; Nitrogen fixation ; Nitrogenase ; Bacteroids ; Symbiosis
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    Notes: Abstract Formate metabolism supported nitrogen-fixation activity in free-living cultures of Rhizobium japonicum. However, formate0dependent nitrogense activity was observed only in the presence of carbon sources such as glutamate, ribose or aspartate which by themselves were unable to support nitrogenase activity. Formate-dependent nitrogenase activity was not detected in the presence of carbon sources such as malate, gluconate or glycerol which by themselves supported nitrogenase activity. A mutant strain of R. japonicum was isolated that was unable to utilise formate and was shown to lack formate dehydrogenase activity. This mutant strain exhibited no formate-dependent nitrogenase activity. Both the wild-type and mutant strains nodulated soybean plants effectively and there were no significant differences in the plant dry weight or total nitrogen content of the respective plants. Furthermore pea bacteroids lacked formate dehydrogenase activity and exogenously added formate had no stimulatory effect on the endogenous oxygen uptake rate. The role of formate metabolism in symbiotic nitrogen fixation is discussed.
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  • 41
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrogenase regulation ; Glutamine synthetase ; Methionine suofoximine ; Rhodospirillaceae
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    Notes: Abstract Methionine sulfoximine (MSX), an irreversible inhibitor of glutamine synthetase of Rhodopseudomonas palustris restored nitrogenase activity to cells in which nitrogenase had been completely inhibited by ammonia switch-off. After addition of MSX, there was a lag period before nitrogenase activity was fully restored. During this lag, glutamine synthetase activity progressively decreased, and near the time of its complete inhibition, nitrogenase activity resumed. Nitrogenase switch-off by ammonia thus required active glutamine synthetase. Glutamine itself caused nitrogenase inhibition whose reversal by MSX depended on the relative ratio of MSX to glutamine. Unlike ammonia, glutamine inhibited nitrogenase under conditions where glutamine synthetase activity was absent. This indicates that glutamine is the effector molecule in nitrogenase switch-off, for instance by interacting with the enzymatic system for Fe protein inactivation. The effects of glutamine and MSX were also dependent on the culture age. Possible explanation for this and for the competitive effects are a common binding site within the regulatory apparatus for nitrogenase, or, in part, within a common transport system. Some observations with MSX were extended to Rhodopseudomonas capsulata and agreed with those in R. palustris.
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  • 42
    ISSN: 1432-072X
    Keywords: Azotobacter vinelandii ; Continuous culture ; Oxygen control ; Nitrogen fixation ; Respiratory protection
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    Notes: Abstract Azotobacter vinelandii strain OP was grown in continuous culture at various dissolved oxygen concentrations of air (100% air saturation of the medium=225 ±14 μM O2). Sucrose was added as carbon source and either dinitrogen or ammonia as nitrogen sources. Irrespective of the nitrogen source steady state cultures showed the following general responses with dissolved oxygen concentrations increasing from about 1% to 30% air saturation: (i) cell protein levels, (ii) the amount of cell protein formed per sucrose consumed as well as (iii) nitrogenase activity decreased by at least a factor of two while (iv) cellular respiration increased. At higher oxygen concentrations the parameters changed only slightly, if at all. Increasing the sucrose concentration in the inflowing medium (s R) from 3 g/l to 15 g/l increased the total level of cellular respiration with nitrogen-fixing cultures but was more pronounced with ammonium-assimilating cultures. With nitrogen-fixing cultures cell protein levels increased five-fold while the ratio of protein formed per sucrose consumed as well as cellular nitrogenase activity remained unaffected. With ammonium-assimilating cultures the cell protein level was only doubled and the level of cell protein formed per sucrose consumed was decreased at the higher s R. Increasing the dilution rate at a constant oxygen concentration of 45% air saturation resulted in an almost parallel increase of both cellular respiratory and nitrogenase activity at low and moderate dilution rates. At high dilution rates nitrogenase activity increased steeply over the respiratory activity. Nitrogen-fixing cultures adapted to various oxygen concentrations were subjected to oxygen stress by increasing the oxygen concentration for 7 min. In all cases, this resulted in a complete inhibition (‘switch-off’) of nitrogenase activity. Upon restoration of the original oxygen concentration nitrogenase activity returned to a decreased level. The discussion arrives at the conclusion that some of the results are incompatible with the concept of respiratory protection of nitrogenase.
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  • 43
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    Archives of microbiology 136 (1983), S. 81-83 
    ISSN: 1432-072X
    Keywords: Ammonia production ; Anabaena ; Cyanobacteria ; Nitrate reductase ; Nitrogen fixation
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    Notes: Abstract In the filamentous heterocystous cyanobacterium Anabaena sp. strain ATCC 33047 dinitrogen fixation and nitrate reduction are mutually exclusive processes. Nitrate promotes nitrate reductase synthesis and represses nitrogenase formation. Inhibition of ammonium assimilation by l-methionine-d,l-sulfoximine (MSX) alleviates the repressive effect of nitrate on nitrogenase synthesis, thus indicating that the nitrate effect is indirect through metabolites generated from the ammonium derived from nitrate reduction. In MSX-treated cells both nitrate reduction and dinitrogen fixation take place simultaneously, although at different sites of the filament, without any apparent competition for the required reducing power. The MSX-treated Anabaena cells generate ammonium from both nitrate and dinitrogen, simultaneously.
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  • 44
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    Archives of microbiology 138 (1984), S. 212-216 
    ISSN: 1432-072X
    Keywords: Denitrification ; Nitrate respiration ; Nitrous oxide reduction ; Nitrogen fixation ; Azospirillum
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    Notes: Abstract Nitrous oxide reduction can consistently be demonstrated with high activities in cells of Azospirillum brasilense Sp 7 which are grown anaerobically in the presence of low amounts of nitrite. Azospirillum can even grow anaerobically with nitrous oxide in the absence of any other respiratory electron acceptor. Nitrous oxide reduction by Azospirillum is inhibited by acetylene, amytal and weakly by carbon monoxide. Azospirillum converts nitrous oxide to molecular nitrogen without the formation of ammonia. The cells must, therefore, be supplied with ammonia from nitrogen fixation during anaerobic growth with nitrous oxide. When no other nitrogen compound besides nitrous oxide is available in the medium, the bacteria synthesize nitrogenase from protein reserves in about 2 h. Nitrogenase synthesis is blocked by chloramphenicol under these conditions. In contrast, the addition of nitrate or nitrite to the medium represses the synthesis of nitrogenase. Nitrous oxide reduction by Azospirillum and other microorganisms is possibly of ecological significance, because the reaction performed by the bacteria may remove nitrous oxide from soils.
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  • 45
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    Archives of microbiology 130 (1981), S. 96-100 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrate respiration ; Denitrification ; Assimilatory nitrate reduction ; Dissimilatory nitrate reduction ; Acetylene reduction ; Azospirillum
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    Notes: Abstract Azospirillum spp. participate in all steps of the nitrogen cycle except nitrification. They can fix molecular nitrogen and perform assimilatory nitrate reduction and nitrate respiration. Culture conditions have been defined under which nitrate is used both as terminal respiratory electron acceptor and as nitrogen source for growth. Nitrate and, possibly to a very limited extent, nitrite, but not sulfate, iron or fumarate support anaerobic respiration. Under anaerobic conditions, nitrate can also supply energy for nitrogen fixation but without supporting growth. Nitrate-dependent nitrogenase activity lasts only for 3–4 h until the enzymes of assimilatory nitrate reduction are synthesized. Nitrite accumulates during this period and inhibits nitrogenase activity at concentrations of about 1 mM.
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  • 46
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    Archives of microbiology 140 (1984), S. 215-217 
    ISSN: 1432-072X
    Keywords: Glutathione reductase ; Cyanobacteria ; Nostoc muscorum ; O2 protection ; Glutathione ; Nitrogen fixation
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    Notes: Abstract Glutathione reductase activity was detected and characterized in heterocysts and vegetative cells of the cyanobacterium Nostoc muscorum. The activity of the enzyme varied between 50 and 150 nmol reduced glutathione· min-1·mg protein-1, and the apparent Km for NADPH was 0.125 and 0.200 mM for heterocysts and vegetative cells, respectively. The enzyme was found to be sensitive to Zn+2 ions, however, preincubation with oxidized glutathione rendered its resistance to Zn+2 inhibition. Nostoc muscorum filaments were found to contain 0.6–0.7mM glutathione, and it is suggested that glutathione reductase can regenerate reduced glutathione in both cell types. The combined activity of glutathione reductase and isocitrate dehydrogenase in heterocysts was as high as 18 nmol reduced glutathione·min-1·mg protein-1. A relatively high superoxide dismutase activity was found in the two cell types; 34.2 and 64.3 enzyme units·min-1·mg protein-1 in heterocysts and vegetative cells, respectively. We suggest that glutathione reductase plays a role in the protection mechanism which removes oxygen radicals in the N2-fixing cyanobacterium Nostoc muscorum.
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  • 47
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    Archives of microbiology 132 (1982), S. 333-337 
    ISSN: 1432-072X
    Keywords: Anabaena variabilis Kütz ; 14C-prelabeled blue-green algae ; Interaction respiration/photosynthesis ; CO2 exchange ; Nitrogen fixation
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    Notes: Abstract Prelabeled Anabaena variabilis Kütz. evolves 14CO2 in the light with KCN and DCMU (2,4-dichlorophenyl-1,1-dimethylurea) present, comparable to the dark control without inhibitors added. Double-reciprocal plots of CO2 release vs. light intensity with either KCN or KCN+DCMU present result in two straight lines intersecting at the ordinate. Apparently, reducing equivalents originating from carbohydrate catabolism are channeled into the photosynthetic electron-transport chain, competing for electrons from photosystem II. Under these conditions, the CO2 release is accompanied by a light-dependent oxygen uptake, presumably due to oxygen-reducing photosystem-I activity while ribulose-bisphosphate carboxylase is inhibited by KCN. Comparing nine blue-green algae it was shown that only nitrogen-fixing species release substantial amounts of CO2 in the light with KCN or KCN+DCMU present. This release is particularly obvious with Anabaena variabilis Kütz. under nitrogen-fixing conditions, but small when the alga is grown with combined nitrogen. We conclude that nitrogen-fixing species share a common link between respiratory and photosynthetic electron transport. The physiological role may be electron supply of nitrogenase by photosystem I.
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  • 48
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    Archives of microbiology 133 (1982), S. 6-10 
    ISSN: 1432-072X
    Keywords: Rhodospirillaceae ; Rhodopseudomonas globiformis ; Nitrogen metabolism ; Nitrogen fixation ; Glutamine synthetase
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    Notes: Abstract Rhodopseudomonas globiformis strain 7950 grew with a variety of amino acids, urea, or N2 as sole nitrogen sources. Cultures grown on N2 reduced acetylene to ethylene; this activity was absent from cells grown on nonlimiting NH 4 + . Glutamate dehydrogenase could not be detected in extracts of cells of strain 7950, although low levels of an alanine dehydrogenase were present. Growth ofR. globiformis on NH 4 + was severely inhibited by the glutamate analogue and glutamine synthetase inhibitor, methionine sulfoximine. High levels of glutamine synthetase (as measured in the γ-glutamyl transferase assay) were observed in cell extracts of strain 7950 regardless of the nitrogen source, although N2 and amino acid grown cells contained somewhat higher glutamine synthetase contents than cells grown on excess NH 4 + . Levels of glutamate synthase inR. globiformis were consistent with that reported from other phototrophic bacteria. Both glutamate synthase and alanine dehydrogenase were linked to NADH as coenzyme. We conclude thatR. globiformis is capable of fixing N2, and assimilates NH 4 + primarily via the glutamine synthetase/glutamate synthase pathway.
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  • 49
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Ultrastructure ; Nitrogen fixation ; Water stress ; Taxonomy ; DNA ; Plasmids
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    Notes: Abstract Two strains of desiccation-tolerant coccoid cyanobacteria, Chroococcus S24, a marine form, and Chroococcus N41, a cryptoendolith isolated from a hot-desert rock, have been characterized. The mol % DNA base compositions of the strains are 47.1 and 48.9% respectively. Plasmid DNA was not detected in either strain. The pigment contents and nutritional characteristics of the strains are identical. Both lack phycoerythrinoid pigments and, in culture, behave as slow-growing halotolerant marine forms with elevated requirements for Na+, Cl−, Mg2+ and Ca2+. Sucrose was the only carbon source of those tested that supported photoheterotrophic growth. Each strain synthesizes nitrogenase under anaerobic conditions but not in air. Morphologically the two strains are indistinguishable. They are considered to be independent isolates of the same cyanobacterial species. Chroococcus N41 was studied in detail with the electron microscope. When brought to equilibrium at matric water potentials of-168 MPa and lower (to-673 MPa=c0.12a w) the protoplast shrinks, but the cells maintain the same size and diameter as those at-2,156 kPa (MN medium; control); the sheath expands and remains attached to the cell wall outer membrane by fibrils. The cell wall, cell membrane, thylakoid membranes, cyanophycin granules and carboxysomes appeared intact in desiccated cells.
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  • 50
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    Theoretical chemistry accounts 60 (1982), S. 579-587 
    ISSN: 1432-2234
    Keywords: Nitrogen fixation ; Nitrogen complexes ; Carbon monoxide complexes ; Electronic effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract MNDO method is used to study the interaction of nitrogen and carbon monoxide molecules with a proton, hydrogen atom, hydride ion, hydrogen molecule ion and hydrogen molecule. Predicted geometries and heats of reaction of different complexes are presented. The wave functions are analyzed in terms of ground state charge distributions and overlap populations. Electronic effects accompanying complexation are also discussed.
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  • 51
    ISSN: 1432-2048
    Keywords: Nitrogen fixation ; Nodule development ; Senescence (nodules) ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cowpea (Vigna unguiculata (L.) Walp cv. Vita 3) seedlings inoculated with Rhizobium strain CB756 were cultured with their root systems maintained in air or in Ar: O2 (80:20, v/v) during early nodule development (up to 24 d after sowing). Compared with those in air, seedlings in Ar:O2 showed progressive N deficiency with inhibited shoot growth, reduced ribulose-1,5-bisphosphate carboxylase and total protein levels and loss of chlorophyll in the leaves. Nodule initiation, differentiation of infected and uninfected nodule tissues and the ultrastructure of bacteriod-containing cells were similar in the air and Ar: O2 treatments up to 16 d after sowing. Thereafter the Ar: O2 treatment caused cessation of growth and development of nodules, reduced protein levels in bacteroids and nodule plant cells, and progressive degeneration of nodule ultrastructure leading to premature senescence of these organs. Provision of NO 3 - (0.1–0.2 mM) to Ar: O2-grown seedlings overcame the abovementioned consequences of N2 deficiency on nodule and plant growth, but merely delayed the degenerative effects of Ar: O2 treatment on nodule structure and senescence. Treatment of Ar: O2-grown seedlings with NO 3 - greatly increased the protein level of nodules but the increase was largely restricted to the plant cell fraction as opposed to the bacteroids. By contrast, NO 3 - treatment of air-grown seedlings increased protein of bacteroid and host nodule fractions to the same relative extents when compared with air-grown plants not supplemented with NO 3 - . These findings, taken together with studies of the distribution of N in nodules of symbiotically effective plants grown from 15N-labeled seed, indicate that direct incorporation of fixation products by bacteroids may be a critical feature in the establishment and continued growth of an effective symbiosis in the cowpea seedling.
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  • 52
    ISSN: 1432-2048
    Keywords: Glutamine synthetase ; Leghaemoglobin ; Nitrogenase ; Nitrogen fixation ; Phaseolus ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activities of glutamine synthetase (GS), nitrogenase and leghaemoglobin were measured during nodule development in Phaseolus vulgaris infected with wild-type or two non-fixing (Fix-) mutants of Rhizobium phaseoli. The large increase in GS activity which was observed during nodulation with the wild-type rhizobial strain occurred concomitantly with the detection and increase in activity of nitrogenase and the amount of leghaemoglobin. Moreover, this increase in GS was found to be due entirely to the appearance of a novel form of the enzyme (GSn1) in the nodule. The activity of the form (GSn2) similar to the root enzyme (GSr) remained constant throughout the experiment. In nodules produced by infection with the two mutant strains of Rhizobium phaseoli (JL15 and JL19) only trace amounts of GSn1 and leghaemoglobin were detected.
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  • 53
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    Cell & tissue research 236 (1984), S. 373-381 
    ISSN: 1432-0878
    Keywords: Merkel cell surface ; Quinacrine fluorescence ; Lectins ; Vibrissae ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Surface carbohydrates on the Merkel cell of the outer root sheath (ORS) were investigated in 1to 4-day-old rat vibrissae by use of rhodamine isothiocyanate (RITC)-conjugated lectins. The red fluorescence of RITC provided a convenient assay for lectin binding to the Merkel cell, which is itself identified by its green fluorescence following selective uptake of the dye quinacrine. In monolayers or suspensions of freshly dissociated ORS cells, the Merkel cell showed high affinity for the α-fucose-specific lectin, Ulex europeus agglutinin I (UEA-I), thus revealing a novel feature for a basally located cell. Other high-affinity lectins included concanavalin A (Con A), wheat germ agglutinin (WGA), soybean agglutinin (SBA), and Ricinus communis agglutinin I (RCA-I). In contrast, Dolichos biflorus (DBA), Bandeiraea simplicifolia I and II (BS-I and BS-II), and peanut agglutinin (PNA) virtually excluded the Merkel cell, though PNA-binding sites were unmasked after neuraminidase treatment. Other dispersed ORS cells had varying lectin affinities, and generally binding was inhibited by a competing haptenic sugar. The pattern of lectin binding seen in cryostat and paraffin sections of the vibrissa suggested that the Merkel cells share surface properties with their neighboring basal and/or spinous cells; however, unshared properties are likely to exist since ingrowing mechanosensory nerves recognize the Merkel cells, and not other epidermal cells, as their targets.
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  • 54
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    Cell & tissue research 236 (1984), S. 711-715 
    ISSN: 1432-0878
    Keywords: Gastric antral mucosa ; Caerulein ; Gastrointestinal hormones ; Cholecystokinin ; Trophic effect ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The growth-promoting effect of caerulein on antral gastric mucosa was explored using Wistar rats. Implanted osmotic minipumps were used to administer submaximal doses of either caerulein or saline to normal rats for up to 4 days. In one group, reflux of bile and pancreatic juice into the stomach was avoided by previous surgical diversion of the distal common bile duct to the jejunum. DNA synthetic and mitotic activity in the antrum epithelium were estimated by 3H-thymidine pulse labelling and autoradiography during the administration of the peptide. The rate of cell migration was determined in animals killed 1, 2 and 3 days after the 3H-thymidine pulse. Administration of caerulein to normal rats provoked significant increases in both labelling and mitotic indices, and a significant acceleration of the upward cell migration in the glandular tubes. In the animals with distal diversion of bile and pancreatic secretions both labelling and mitotic indices were also increased over control values under the effect of the peptide. These data indicate that administration of caerulein stimulates cell proliferation in the antral gastric mucosa. This effect cannot be explained through increased reflux of pancreaticobiliary secretions in the stomach.
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  • 55
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    Cell & tissue research 236 (1984), S. 699-709 
    ISSN: 1432-0878
    Keywords: Testis ; Spermatogenic cycle ; Sertoli cell ; Lipid ; Morphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The volume and surface area of lipid inclusions often present in the cytoplasm of rat Sertoli cells was measured directly from semi-thin sections of perfusion-fixed testicular tissues using an image analyser linked to a light microscope. Sertoli cell nuclei were used as a reference for comparing any variations in the measured parameters of lipid inclusions during the rat spermatogenic cycle. Volume density of Sertoli cell lipid inclusions was assessed by morphometric analysis of Sertoli cells photographically reconstructed from electron micrographs. Maximum lipid content in Sertoli cells occurred during stages IX–XIV of the spermatogenic cycle, then declined at stages I–III and remained low from stages IV–VIII. The persistence and increase in number of many large Sertoli cell lipid inclusions beyond the stage where spermatid residual bodies are phagocytosed within the Sertoli cells (stage IX) suggests that the synthesis and lipolysis of Sertoli cell lipid inclusions represents an intrinsic functional cycle of the Sertoli cells. Stage-dependent variations in the lipid content of rat Sertoli cells offers morphological evidence that the metabolic duties of the Sertoli cells are synchronised with the spermatogenic cycle to provide local coordination of the proliferation and maturation of the germ cells.
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  • 56
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    Cell & tissue research 236 (1984), S. 717-724 
    ISSN: 1432-0878
    Keywords: Ovary ; Ovarian follicle ; Atresia ; Immunoregulation ; Immune tolerance ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Thy-1+ cells, producing Thy-1+ material, have been demonstrated by the indirect immunoperoxidase technique in the theca of growing ovarian follicles of the rat. OX-2 antigen, known as the minor glycoprotein of rat thymocytes, was detected in granulosa cells of non-growing follicles. Ia+ cells of dendritic type and/or activated macrophages were identified in the granulosa of advanced degenerating follicles, and remnants of the zona pellucida exhibited immunoglobulins. In some ovaries immunoglobulins were also bound to the zona pellucida of oocytes of early degenerating antral follicles. Medium-sized antral follicles with degenerating granulosa were occasionally invaded by cells carrying antigens of cytotoxic T lymphocytes or other T lymphocyte subsets, while degenerating large antral follicles were sometimes invaded by cells exhibiting antigen of cells with natural killer function (but not antigens of T lymphocytes). Granulosa cells of some degenerating antral follicles exhibited class-I antigens derived from the major histocompatibility complex. We suggest that cell-mediated control mechanisms of antigen expression and metabolism of tissue cells during their differentiation and degeneration should be considered in addition to the well-documented hormonal dependence of some tissues.
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  • 57
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    Cell & tissue research 237 (1984), S. 245-252 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Interstitial cells ; Astrocytes ; Immunocytochemistry ; Rat ; Mouse
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    Topics: Biology , Medicine
    Notes: Summary Antigenic markers characteristic of astrocytes and their differentiative states (i.e., glial fibrillary acidic protein (GFAP), vimentin, and M1 and C1 antigens) were investigated in the pineal gland of mouse and rat using double immunolabeling techniques. In both species the socalled interstitial cells as characterized by TEM were shown to be astrocytes, since they expressed vimentin, but neither fibronectin (a marker for fibroblasts and endothelial cells) nor the neuron-specific L1 antigen or tetanus toxin receptors. Subpopulations of vimentin-positive pineal astrocytes were also GFAP- and C1- antigen-positive. M1- antigenpositive cells were not detected. It is concluded that a considerable proportion of interstitial cells in the pineal gland of rat and mouse are immature astrocytes which, in contrast to other parts of the central nervous system, persist into adulthood.
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  • 58
    ISSN: 1432-0878
    Keywords: Thyroid gland, fetal ; Cytoskeleton ; Cytocha lasin B ; Vinblastine ; Colchicine ; Follicular development (thyroid) ; Tissue culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Thyrotropic hormone (TSH) or cAMP accelerate the formation of follicular cavities in the explanted thyroid gland of the 15-day-old rat fetus. Cytochalasin B or vinblastine and nocodazole or colchicine, which disorganize microfilamental and microtubular structures respectively, inhibit or completely block in vitro-induced folliculogenesis. Exposure of the thyroid tissue to lumicolchicine, a structural isomer of colchicine deprived of antimicrotubular activity, does not inhibit the activation of folliculogenesis induced by TSH. These results are strong evidence for the supposition that microfilaments and microtubules are involved in the TSH-stimulated mechanisms resulting in thyroid folliculogenesis. Folliculogenesis requires the integrity of both microfilaments and microtubules.
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  • 59
    ISSN: 1432-0878
    Keywords: CRF-neurons ; Hypothalamus ; Development, ontogenetic ; Rat
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    Topics: Biology , Medicine
    Notes: Summary Appearance of immunoreactive corticotropin-releasing factor (CRF)-containing neurons was studied in developing hypothalamus of the rat by use of antisera against rat- and ovine CRF. These neurons were first recognized in the lateral and paraventricular nuclei on days 15.5 and 16.5 of gestation, respectively, when antiserum against rat CRF was employed. Antiserum against ovine CRF revealed the cells two days later exclusively in the latter nucleus. In both nuclei, the neurons increased in number with development. The neurons in the paraventricular nucleus appeared to project their immunoreactive processes to the median eminence via the periventricular and lateral pathways. In the median eminence, the immunoreaction with antiserum to rat CRF was first recognized in its anterior portion in the form of dots on day 16.5 of gestation but as beaded fibers in the external layer on day 17.5; these structures increased in amount with development in rostro-caudal direction. Although antiserum to ovine CRF was less potent in immunostainability than antiserum to rat CRF, it also revealed the beaded fibers in the median eminence on day 17.5 of gestation. Since evidence is available that the paraventricular nucleus is involved in corticotropin release, it is concluded that, in rats, the hypothalamic regulatory mechanism controlling the release of corticotropin initially appears on days 16.5–17.5 of gestation.
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  • 60
    ISSN: 1432-0878
    Keywords: Oxytocin ; Paraventricular nucleus ; Supraoptic nucleus ; Anterior commissural nucleus ; Immunohistology ; Rat
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    Topics: Biology , Medicine
    Notes: Summary The effect of water deprivation or estrogen treatment on the oxytocin content of rat hypothalamic cells was examined using a quantitative immunohistological technique. Oxytocin-containing cells were visualized using the immunoperoxidase technique of Sternberger and a primary antiserum directed against oxytocin. The optical density of the darkest 3.2 μm diameter spot in the cytoplasm of a cell was used as a measure of the oxytocin content of that cell. Water deprivation produced a significant decrease in anti-oxytocin staining in the anterior commissural nucleus of males and females. There was a similar decrease in the paraventricular nucleus of males, but not in the paraventricular nucleus of females or the supraoptic nucleus of either males or females. Estrogen treatment of ovariectomized female rats produced a fall in anti-oxytocin staining in the anterior commissural, but not paraventricular or supraoptic nuclei.
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  • 61
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    Cell & tissue research 205 (1980), S. 11-17 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Subcommissural organ ; Vasopressin ; Oxytocin ; Vasotocin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vasopressin and oxytocin were specifically demonstrated in the rat brain using the unlabelled antibody-enzyme method and purification of the first antiserum. Vasopressin and oxytocin fibres extend via the subcommissural organ or habenular commissure into the pineal stalk and terminate in the anterior part of the pineal organ. In addition, immediately adjacent to the subsommissural organ many vasopressin-containing fibres run caudally toward the central grey. These results are discussed in relation to the proposed presence of vasotocin in the pineal gland.
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  • 62
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    Cell & tissue research 210 (1980), S. 167-170 
    ISSN: 1432-0878
    Keywords: Neuropeptides ; VIP-immunoreactive neurons ; Retina ; Amacrine cells ; Rat
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    Topics: Biology , Medicine
    Notes: Summary Immunoreactive vasoactive intestinal polypeptide (VIP) was detected in a population of amacrine cells in the retina of the rat. Processes of these cells reach both the inner and outer half of the inner plexiform layer where they form sublayers. The VIP neurons are different from previously known amacrine cell types.
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  • 63
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    Cell & tissue research 210 (1980), S. 181-189 
    ISSN: 1432-0878
    Keywords: Catecholamines ; Neurophysin ; Simultaneous demonstration ; Functional interaction ; Rat
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    Topics: Biology , Medicine
    Notes: Summary A method was developed that allows the analysis of neuropeptides and monoamines in a single tissue section by the application of the unlabeled antibody method for peptide staining to tissue sections freeze-dried for formaldehyde-induced monoamine histofluorescence. The hypothalamic magnocellular system of male albino rats served as a model for this study; neurons were stained with anti-neurophysin sera, which mark the vasopressin- and oxytocin-associated proteins. Neurophysin-containing perikarya appeared to be surrounded by catecholamine-containing varicosities. This phenomenon was seen to varying degrees within the supraoptic and paraventricular nuclei. The juxtaposition of varicosities and peptidergic neurons suggests an afferent fiber-target neuron relationship that might favor a functional interaction between monoamines and neuropeptides.
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  • 64
    ISSN: 1432-0878
    Keywords: Rat ; Preovulatory follicle ; Ultrastructure ; Degeneration ; Atresia
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    Topics: Biology , Medicine
    Notes: Summary To identify and describe ovarian follicles committed to undergo follicular degeneration (atresia), immature rats were primed with pregnant mare serum gonadotropin (PMSG). After PMSG treatment, preovulatory follicles develop but subsequently degenerate. Prior to the appearance of pyknotic nuclei (Stage I of atresia), degenerative changes were observed in focal areas of the granulosa cell layer. These changes include “blebbing” of the cytoplasm and alterations in the shape of the granulosa cells. The appearance of these degenerative changes coincides with a decrease in ovarian concentrations of estradiol and testosterone. Since estrogens and androgens maintain the follicle, the decline in estradiol and testosterone could be responsible for the further degenerative alterations that lead to complete deterioration of the preovulatory follicle. In Stage I atretic follicles, lysosome-derived autophagic vacuoles develop and macrophages invade both the thecal and granulosa cell layers. The combined actions of the autophagic vacuoles and macrophages could destroy both the granulosa-cell and thecal layers and thereby transform the preovulatory follicle into an ovarian cyst.
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  • 65
    ISSN: 1432-0878
    Keywords: Lymph node ; Macrophages ; Postcapillary high endothelial venules ; Rat
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    Notes: Summary The afferent lymphatic vessels of rat popliteal lymph nodes were interrupted, and the histological alterations in the lymph nodes occurring 1 to 14 weeks after operation were studied. One week after operation the number of macrophages was considerably reduced and continued to decrease during the subsequent time periods studied. A 6 weeks most macrophages had disappeared. Simultaneously the immunological activity diminished and had completely disappeared 8 weeks after operation. Three weeks after operation the endothelial cells of the postcapillary high endothelial venules had flattened, and the number of immigrating lymphocytes was greatly reduced. Subsequently the lymph nodes became depleted of both macrophages and lymphocytes, leaving only the reticuloendothelial framework.
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  • 66
    ISSN: 1432-0878
    Keywords: Adrenalectomy ; Hypophysectomy ; Compensatory adrenal growth ; Rat ; Stereology
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    Topics: Biology , Medicine
    Notes: Summary Compensatory hypertrophy of the remaining gland in unilaterally adrenalectomized rats was investigated by morphometric techniques. It was observed that compensatory adrenal growth occurred in both dexamethasone-treated and hypophysectomized rats, receiving maintenance doses of ACTH. However, it was only half that found in intact animals. These results support the view that activation of the hypothalamo-hypophyseal axis is not the unique mechanism underlying adrenal compensatory hypertrophy in the rat.
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  • 67
    ISSN: 1432-0878
    Keywords: Caldesmon ; Actin ; Immunocytochemistry ; Small intestine ; Smooth muscle ; Rat
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    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin-interacting protein) (Sobue et al. 1982) and of actin was studied in the rat's small intestine by means of light-microscopic immunocytochemistry. Positive immunostaining for caldesmon was seen in smooth muscle cells of the intestinal wall, and of blood vessels, and in the apical portion of the absorptive epithelial cells. The immunoreactivity in goblet cells was difficult to recognize. The positive reaction to immunostaining for actin showed almost the same pattern as that for caldesmon. These results suggest that this calmodulin-binding protein may play an important role in the control of actin-myosin interaction in smooth muscle cells and in non-muscle cells.
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  • 68
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    Cell & tissue research 235 (1984), S. 433-438 
    ISSN: 1432-0878
    Keywords: Mammary gland ; Ferritin-concanavalin A ; Concanavalin A ; Endocytosis ; Membrane reuse ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ferritin-concanavalin A (Fer-Con A) was used to label the apical plasma membrane of the lactating cell to determine whether membrane internalization takes place. Rat glands were infused in vivo via the teat with 0.2 mg of Fer-Con A in 0.2 ml tris buffer (pH 7.0) containing 0.1% trypan blue, the latter acting as a marker of the infusate. Tissues were obtained from separate animals 5, 10 and 60 min postinfusion. Fer-Con A was seen in alveolar lumina bound to the outer surfaces of apical plasma membrane, microvilli and milk fat globules. It was observed within lactating cells on the inner membrane surfaces of endocytotic vesicles, Golgi cisternae, and secretory vesicles containing casein micelles, and in multivesicular bodies and lysosomes. Internalization of the ferritin-lectin conjugate into casein-containing secretory vesicles was detectable in the 5-min postinfusion tissue. Lysosomes were the only structures in control tissue that contained particles bearing some resemblance to Fer-Con A. The data provide evidence that apical plasma membrane is internalized and distributed to a number of intracellular compartments.
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  • 69
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    Cell & tissue research 235 (1984), S. 449-452 
    ISSN: 1432-0878
    Keywords: Suprachiasmatic nucleus ; Morphometry ; Synapses ; Sexual dimorphism ; Rat
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    Topics: Biology , Medicine
    Notes: Summary The suprachiasmatic nucleus (SCN) of male rats was estimated to contain 16×106 synaptic appositions (unilaterally) or 250×106 appositions in 1 mm3 tissue of the nucleus with an average of 1404 appositions per neuron. There are significantly fewer synaptic appositions in the suprachiasmatic nucleus of female rats (15×106 per SCN or 236×106 in 1 mm3 tissue of SCN with 1264 appositions per neuron on an average). Additionally, numbers of various types of synapses (axo-somatic, invaginated, dendrodendritic and optic) are estimated.
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  • 70
    ISSN: 1432-0878
    Keywords: Monosodium-1-glutamate ; Neuropathology ; Rat ; Superior colliculus ; Toxicology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Systemic administration of monosodium-1-gluta-mate by single injections of 4 mg/g body weight in infant rats (2–10 days of age) results in acute swelling of cytoplasm and nuclear pyknosis of neurons in the stratum zonale and stratum griseum superficiale of the superior colliculus. Multiple daily doses of 4 mg/g body weight monosodium-1-glutamate result in an almost complete loss of neurons in these two superficial layers. The deeper layers appear not to be affected. No pathological effects were observed in the lateral geniculate body or pretectal complex. Light-and electron-microscopic studies reveal that the optic nerves are remarkably shrunken and many myelinated as well as unmyelinated axons are lost. Injection of 3Hproline into the vitreous body of one eye results in limited transport to the suprachiasmatic nucleus, lateral geniculate body and to lateral portions of the superior colliculus. The small percentage of intact axons in the optic nerve, as well as the limited proline transport from the eye, suggest that administration of monosodium-1-glutamate leaves intact some optic fibers, a portion of which belongs to the retinohypothalamic tract.
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  • 71
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Calcium pool ; Calcium release ; Electron microscopy ; Rat
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    Notes: Summary In an attempt to identify a cellular Ca2+-pool, from which calcium is released when secretagogues are applied, tissue fragments of the rat exocrine pancreas were incubated and fixed with glutaraldehyde in the presence of calcium. By means of this procedure electron-dense deposits were found on plasma membranes. X-ray microanalysis showed that these deposits contain calcium. Stimulation of tissue fragments with the use of the secretagogues carbachol or cholecystokinin reduced the number of deposits by about 80%. When the antagonist atropine was applied after carbachol stimulation, deposits reappeared on cell membranes, which then disappeared again after a second stimulation with cholecystokinin. In the presence of procaine, carbachol was inhibited and only slightly reduced the Ca2+-deposits on the plasma membranes. These results suggest that a calcium pool, from which calcium is released to induce enzyme secretion on stimulation, is located in the cell membrane
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  • 72
    ISSN: 1432-0878
    Keywords: Somatostatin ; Ontogenesis ; Electron-microscopic immunohistochemistry ; Median eminence ; Rat
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    Notes: Summary The development of immunoreactive (ir) somatostatin-containing nerve terminals in the rat median eminence (ME) has been examined electron-microscopically. Nerve fibers containing ir particles scattered throughout the axoplasm are first seen in the external layer of the ME on day 18.5 of gestation, and, on day 21.5 appear to terminate on the basement membrane of the perivascular space of the portal vessels. After birth, the fiber terminals contain several membrane-limited granules, which are labeled with ir PAP particles. Ultrathin, Epon-embedded sections of ME, treated by the protein A gold-labeling method for somatostatin, demonstrate positively labeled granules in the nerve fibers in the postnatal ME, but in the prenatal tissue, no specific gold-labeling is found. These findings show that, in the external layer of the ME, somatostatin storing occurs in the granules in the axonal terminals after birth.
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  • 73
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    Cell & tissue research 236 (1984), S. 491-493 
    ISSN: 1432-0878
    Keywords: Brain vessels ; Basal lamina ; Pericytes ; Endothelial cells ; Glial cells ; Argyrophilic staining ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vibratome sections obtained from perfusion-fixed rat brains were stained by means of silver impregnation and physical development according to Gailyas (1970). Small pieces of the cerebral cortex were postfixed with buffered osmium tetroxide solution and processed for electron microscopy to examine the localization of the silver deposit at the cellular level. The cell surfaces of pericytes and smooth muscle cells were completely outlined by silver grains. Endothelial cells and perivascular astrocytes, however, showed an asymmetric distribution of the silver deposit, i.e., the deposit was restricted to the abluminal endothelial surface and to the astrocytic membrane adjacent to the vessel wall, respectively. The method allowed a clear-cut distinction between perikarya of endothelial cells and pericytes as well as glial cells in perivascular position, even at the light-microscopic level.
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  • 74
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    Cell & tissue research 236 (1984), S. 249-255 
    ISSN: 1432-0878
    Keywords: Oocyte ; Nucleolus ; Silver staining ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The nucleoli of dictyate-stage growing oocytes in rat ovaries were examined both with routine electron microscopy and electron microscopy after silver nitrate and ammoniacal silver nitrate (Ag-AS) staining. The nucleoli of the unilaminar follicular oocytes consist of twisted strands of dense fibrillar components, aggregates of granular components, and small fibrillar centers. After Ag-AS staining, silver grains are numerous on the dense fibrillar strands, fewer on the fibrillar centers, and very sporadic on the granular aggregates. The same stainability of three nucleolar components with the Ag-AS method was also confirmed in the nucleoli segregated by actinomycin D. During the transition of growing oocytes from bilaminar to plurilaminar follicle stage, the nucleolar dense fibrillar strands gradually conglomerate and are transformed into large and compact spherules. The stainability of dense fibrillar components with the Ag-AS method was lost along with this nucleolar transformation. These results may provide some new clues on the functional significance of AgAS-positive proteins in the nucleoli.
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  • 75
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    Cell & tissue research 237 (1984), S. 103-109 
    ISSN: 1432-0878
    Keywords: Synapses ; Synaptogenesis ; Development fetal ; Olfactory cortex ; Rat
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Electron microscopy was used to study synaptogenesis in prepyriform cortex of fetal rat pups during early stages of synapse formation. Of special interest is the frequent occurrence of unapposed, developing synaptic specializations in axon and growth cone profiles. The location and morphology of the unapposed specializations suggests that thay are presynaptic in nature. These presumably immature presynaptic specializations are found in the lateral olfactory tract and subjacent cortex. Intermediate forms between uncontacted presynaptic specializations and definitive synapses suggest a synaptogenic sequence in which initial development of an immature presynaptic specialization begins without apposition of a postsynaptic element at that location. This implies that initiation of presynaptic development is not dependent upon postsynaptic contact and also raises the question of whether synaptic contacts could be established via presynaptic induction of postsynaptic formation.
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    Cell & tissue research 237 (1984), S. 185-186 
    ISSN: 1432-0878
    Keywords: Gap junction ; Cytoskeleton ; Heart ; Ultrarapid freezing ; Rat
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Using ultrarapid-freezing techniques and freezefracture electron microscopy, we report here a close association between cardiac gap junctions and specialized membrane domains containing regularly-spaced furrows. These specialized furrowed domains are observed only during periods of gap junction re-organisation (i.e., connexon redistribution) and may reflect the presence of underlying cytoskeletal elements controlling the position of connexons in the membrane.
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  • 77
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    Cell & tissue research 237 (1984), S. 371-373 
    ISSN: 1432-0878
    Keywords: Endothelium ; Transport, intracellular ; Transport vesicles, channels ; Micropinocytosis ; Capillaries ; Endometrium ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Three types of transendothelial channels are described in the endothelium of blood capillaries in the endometrium of the rat. It is postulated that they may function as pores draining interstitial fluid to the venous blood.
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  • 78
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    Keywords: Thyroid ; Immunocytochemistry ; Caldesmon ; Actin ; Endocytosis ; Rat
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    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin interacting protein; Sobue et al. 1982) and actin was studied in the rat thyroid gland by means of light-microscopic immunocytochemistry, and the fine-structural distribution of actin filaments was examined by use of heavy meromyosin (HMM). Caldesmon and actin were demonstrated in the apical cytoplasm of almost all the follicle epithelial cells in normal as well as TSH-treated animals. Immunoreactivities for both caldesmon and actin showed almost the same pattern in localization. The smooth muscle cells of the blood vessels were also positive for caldesmon and actin. By electron microscopy, numerous actin filaments decorated by HMM and running perpendicularly or randomly to the apical surface were recognized in the apical cytoplasm of the follicle epithelial cell. These results suggest that caldesmon and actin, in conjugation with calmodulin, play a role in the regulation of cellular activity such as exocytosis and endocytosis in the apical portion of the follicle epithelial cell.
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  • 79
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    Cell & tissue research 209 (1980), S. 1-10 
    ISSN: 1432-0878
    Keywords: Myoepithelial cell ; Exocrine gland ; Scanning electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By removing connective tissue components with enzymatic digestion followed by HCl-hydrolysis, myoepithelial cells (MECs) of the terminal portion in a variety of exocrine glands of the rat were examined with the scanning electron microscope. The profile of MECs varied considerably from gland to gland; MECs in the lactating mammary gland have a few long cytoplasmic processes in close contact with those of adjacent cells forming a continuous network around the terminal portion. Those of the exorbital lacrimal gland are stellate with many thin radiating processes with tapered ends that terminate freely. MECs in the sublingual gland are characterized by a number of broad and extensive cellular processes. MECs in the submandibular gland are similar in appearance to those of the exorbital lacrimal gland, but with more extensive cellular processes that form a more or less continuous network with those of the adjacent cells. No MECs were observed on the terminal portion of the parotid gland where the cells appear to be lodged on the intercalated duct. The relative surface area covered by MECs per terminal portion was also found to vary significantly, being 24% in the lactating mammary, 17% in the exorbital lacrimal, 48% in the sublingual, and 25% in the submandibular glands. The findings are discussed in relation to the physical properties of secretions in different glands.
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  • 80
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    Cell & tissue research 207 (1980), S. 89-107 
    ISSN: 1432-0878
    Keywords: Rehydration ; Nongranular vasopressin ; Intercellular clefts ; Axoplasmic reticulum ; Ultrastructural immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The vasopressin system of the rat was examined in the course of the first 12 h of rehydration after prolonged thirst at light and electron microscopic levels and by use of the peroxidase anti-peroxidase (PAP) method. Light microscopically, the median eminence was the only part of the system that not only displayed distinct differences between animals of different rehydration times but also showed a characteristic pattern of immunohistochemical reactivity in its rostro-caudal distribution. Ultrastructurally, in the perikarya a maximal labeling of the rough endoplasmic reticulum was observed after 2 h of rehydration, whereas an extensive labeling of the enlarged Golgi zones was attained after 4 h of resupplying water. A labeling of the intercellular clefts in the basal glial labyrinth of the supraoptic nucleus (and to a lesser degree in the subependymal neuropil adjacent to the paraventricular nucleus) was increased 30 min after the onset of drinking, as compared with water-deprived animals; it decreased slightly after 12 h of rehydration. The filling of the swollen fibers by increasing amounts of labeled axoplasmic reticulum, evident in the nuclear areas already after 30 min of water supply, begins in the median eminence after 2 h of rehydration and is fully developed after 4 and 8 h. Corresponding results hold true for the neural lobe but are somewhat delayed in comparison to the findings in the median eminence. The discussion considers (i) synthesis and transport of nongranular vasopressin within the axoplasmic reticulum, and (ii) release not only from the neural lobe but also from the nuclear areas and from the fibers of the median eminence.
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  • 81
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    Cell & tissue research 233 (1983), S. 485-505 
    ISSN: 1432-0878
    Keywords: Liver ; Pit cell ; Cytoplasmic granules ; Vesicles ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pit cells, on which almost no further contributions have been presented since the first report by Wisse et al. (1976), are described in detail in the rat liver. These cells show several characteristic features: 1) “rod-cored vesicles”, a new type of vesicular inclusion observed first in our study; 2) electron-dense granules, which we consider to arise from multivesicular bodies by the accumulation of dense material; and 3) well-developed pseudopodia. Although these features clearly differentiate pit cells from conventional lymphocytes, these two cell types display similarities (i) in a number of ultrastructural features, (ii) in the pattern of their intralobular distribution, and (iii) in their presence in the spleen and peripheral blood.
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  • 82
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    Cell & tissue research 233 (1983), S. 689-691 
    ISSN: 1432-0878
    Keywords: Fluoride ; Progeny ; Bone ; Rat
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    Topics: Biology , Medicine
    Notes: Summary Femurs of weanling rats from third-pregnancy litters of dams given 150 ppm fluoride in the drinking water were examined by light- and scanning-electron microscopy. Under the conditions of the experimental procedure, no pathological changes were seen in the femurs as a result of maternal ingestion of fluoride. These results indicate that the amount of fluoride crossing the placenta and mammary gland was insufficient to produce significant morphological changes in the bones of 3-week-old rats.
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  • 83
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    Cell & tissue research 234 (1983), S. 125-134 
    ISSN: 1432-0878
    Keywords: Vasopressin ; Immunocytochemistry ; Electron microscopy ; Hypothalamus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vasopressin-containing neurons, identified by immunocytochemistry, are located predominantly in the posterior magnocellular division of the paraventricular nucleus of the rat hypothalamus. By electron microscopy, the immunoreaction product is seen within the cell bodies and neuronal processes. In the perikarya and dendritic processes, the immunoreactive material is associated primarily with neurosecretory granules. Axonal processes, identified by their content of microtubules and accumulation of neurosecretory granules, show the immunoreaction product in association with both of these organelles. Afferent axo-dendritic, axo-somatic and putative axo-axonic synapses with immunostained vasopressinergic neurons can be identified. The presynaptic profiles do not contain immunoreactive material. This study contributes to the ultrastructural characterization of vasopressinergic neurons in the paraventricular nucleus and of their afferent synaptic input.
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  • 84
    ISSN: 1432-0878
    Keywords: Hypothalamo-hypophysial system ; Capillaries ; Ontogenesis ; Rat
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    Notes: Summary The maturation of the capillaries of the primary portal plexus in rats during the perinatal period has been studied lightand electronmicroscopically. The number of capillaries covering the median eminence and of those invading the nervous tissue (capillary loops) increases significantly with age. Capillary loops were observed as early as the 18th fetal day. The mitotic divisions of the endothelial cells within the preexisting capillaries seem to be the main reason for the vascular growth. Immature capillaries with a characteristic narrow lumen are surrounded by a fuzzy basal lamina; their wall is formed by a generally expanded endothelium with rather sparse organelles and inclusions, and by minute flattened areas. The maturation of the capillary results in a progressive spread of flattened endothelium followed by an enlargment of the capillary lumen. Moreover, a rising concentration of organelles and inclusions, relatively numerous luminal microvilli, and a dense and uniform basal lamina become noticeable as capillary differentiation proceeds. These data are thought to reflect the progressive increase in the metabolic activity of the endothelium as well as the establishment of capillary patency during the perinatal period of rats.
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  • 85
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    Cell & tissue research 235 (1984), S. 187-194 
    ISSN: 1432-0878
    Keywords: Golgi apparatus ; Monensin ; Small intestine ; Cytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of short-time treatment with the ionophore monensin, administered intraluminally at concentrations of 5 and 10 μM, was studied on the Golgi apparatus of absorptive cells in the small intestine of the rat. At 2–3 min after treatment most of the Golgi stacks exhibited dilated cisternae. At 4–5 min stacked cisternae were absent; they were replaced by groups of smooth-surfaced vacuoles. Dilatation and vacuolization occurred in the entire stacks without preferential effect on any particular Golgi subcompartment. Monensin did not influence the cytochemical Golgi reaction of thiamine pyrophosphatase and acid phosphatase. The characteristic staining pattern of these two enzymes in all Golgi cisternae of absorptive cells in the proximal small intestine, and the reactivity restricted to trans cisternae in distal segments of the small intestine, were unchanged after treatment with monensin. In the distal small intestine, the cytochemical pattern allowed the monensin-induced vacuoles to be attributed to the former cisor trans-Golgi face. Further, the cytochemical results demonstrate that vacuolization is not restricted to the stacked cisternae, but includes the trans-most cisterna. The latter, usually located at some distance from the Golgi stacks, has been defined as belonging to the GERL system in several types of cells. The clear response to monensin, an agent that selectively affects the Golgi apparatus, indicates common properties between trans-most and stacked Golgi cisternae.
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  • 86
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    Cell & tissue research 235 (1984), S. 485-489 
    ISSN: 1432-0878
    Keywords: Ventromedial nucleus ; Hypothalamus ; Ultra-structure ; Nucleoli ; Estrogen effects ; Chromatin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Estrogen is accumulated from the blood by nerve cells in the ventromedial nucleus of the hypothalamus and can facilitate female reproductive behavior by acting on this region of the brain. This cell group was examined in ovariectomized female rats, given estrogen or control treatment, by use of light and electron microscopy. A significantly greater portion of the nerve cells in the estrogen-treated animals had protuberances on their nucleolar surfaces, apparent under the light microscope. The fine structure of such protuberances included dense, aggregated material, which is shown to contain DNA by the sodium tungstate staining technique. Because increased numbers of such protuberances were found in nuclei of cells of the experimental group where previous studies demonstrated a significant increase in ultrastructural signs of biosynthetic activity, they may be associated with increased RNA synthesis. Thus, they could indicate, ultrastructurally, increased synthetic rates for RNA in nerve cells through which estrogen promotes reproductive behavior.
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  • 87
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    Cell & tissue research 235 (1984), S. 669-673 
    ISSN: 1432-0878
    Keywords: Liver-cell heterogeneity ; Hepatic venous branches ; Karyometry ; Binucleate cells ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In untreated adult male albino rats nuclear volume and the percentage of binucleate cells were determined in the first layer of hepatocytes adjacent to hepatic venous branches of varying diameters (〈40 μm, 40 μm–80 μm, 80 μm–120μm, 120 μm–160 μm, 〉160 μm), and in the third and fourth layer of hepatocytes in the remainder of the perivenous parenchyma. In the first layer of hepatocytes adjacent to the vascular structures means of nuclear volume are significantly lower and percentage of binucleate cells significantly higher than in the cells of the remainder of the perivenous parenchyma. Within each area measured distribution curves of nuclear volume classes were homogeneous but showed heterogeneity in comparison with each other. The morphometric data presented in this study strongly support the opinion of the heterogeneity of liver cells in the perivenous zone, as previously postulated on the basis of histochemical investigations.
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  • 88
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    Cell & tissue research 236 (1984), S. 305-315 
    ISSN: 1432-0878
    Keywords: Hepatocytes ; Rat ; Liver ; Circadian rhythm ; Morphometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Subcellular structures of hepatocytes in periportal and perivenous zones were examined during 24 h. The volume, surface and numerical profile densities of cytoplasmic organelles were analysed morphometrically. Most subcellular structures in periportal and perivenous hepatocytes were subject to strong circadian variations. In hepatocytes from both zones, the volume densities of smooth endoplasmic reticulum (sER), mitochondria, lysosomes, peroxisomes, polysomes and lipid droplets demonstrated peak values at 16.00 h, 20.00 h or 00.00 h; trough values were at 04.00 h, 08.00 h, or 12.00 h, except for peroxisomes (16.00 h). However, the volume densities of glycogen granules and rough endoplasmic reticulum (rER) in periportal and perivenous hepatocytes exhibited maximal values at 04.00 h, 08.00 h or 12.00 h and minimal values at 20.00 h. The surface densities of sER, mitochondria, lysosomes and peroxisomes, and the numerical profile densities of mitochondria, lysosomes and peroxisomes in periportal and perivenous hepatocytes showed similar trends. These events suggest that membranes of the rER show a partial correlation with the sER, mitochondrial and lysosomal membranes during the 24-h span. This may involve the interaction between ribosomes and rER. Almost all cytoplasmic organelles examined displayed significant differences between periportal and perivenous hepatocytes, morphometrically and in fine structure, indicating that the morphofunctional variability of hepatocytes differs depending on the location in the liver acinus.
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  • 89
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    Cell & tissue research 236 (1984), S. 321-325 
    ISSN: 1432-0878
    Keywords: Triiodothyronine ; Radioautography ; Mitochondria ; Liver ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To assess the distribution of the thyroid hormone triiodothyronine (T3) within intact living cells, freshly prepared dispersed rat hepatocytes were incubated with [125I]-T3 for periods of 5 min and 30 min. Lightand electron-microscopic (EM) radioautography was carried out to determine the distribution of grains over the isolated cells. Both procedures showed the grains distributed almost entirely over the cytoplasmic matrix rather than the nucleus. Grain counts under the EM were compared with expectation based on established quantitative methods. Only the mitochondria showed obvious and statistically significant grain counts, whereas the nucleus failed to accumulate grains in excess of expectations by chance alone based on area. The findings support the existence of mitochondrial binding of T3, presumably a prerequisite for its action in direct stimulation of the mitochondria.
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  • 90
    ISSN: 1432-0878
    Keywords: Lymph node ; Steroids ; Macrophages ; Intercellular junctions ; Rat
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    Topics: Biology , Medicine
    Notes: Summary Intercellular junctions were often found between macrophages in sinuses of regional lymph nodes of the rat after injection of large doses of cholesterol, cortisone acetate, and estrone at the footpad. They were identified by subplasmalemmal densities, 20–50 nm in width, beneath the plasma membranes of apposed macrophages. No distinct filamentous structures were visible in those dense regions. Electron-dense amorphous materials are lined up at the center of the intercellular space in the junctional regions. Some macrophages form clusters with intercellular junctions. No significant difference in the effect of cholesterol, cortisone acetate, and estrone on the number of intercellular junctions betwene macrophages was found.
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  • 91
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    Cell & tissue research 236 (1984), S. 171-180 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ultrastructure ; Supraoptic nucleus ; Neuropil ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neuropil located ventral to the SON was investigated by the use of immunoperoxidase staining for neurophysins, oxytocin and vasopressin, and electron miroscopy. The study was performed in six groups of rats: 1) control; 2) infusion of isotonic saline into the CSF; 3) infusion of hypertonic saline into the CSF; 4) drinking hypertonic saline for 4 days; 5) same as group 4 but injection of colchicine into the CSF on second day of dehydration; 6) salt loading for 3 months. In the control rats the ventral neuropil contained a few immunoreactive processes, the general morphology of which was completely different from that of the neurosecretory axons emerging from the SON at its dorsal aspect. In rats of groups 3 to 6 the ventral processes (VP) became loaded with neurosecretory granules, whereas the perikarya and axons were depleted. Based on their general morphology and reactivity pattern it is suggested that the VP are dendrites. Most of these “dendrites” were embedded in a glial cushion formed by the processes of a particular type of marginal glia. Some of these “dendrites” enveloped an arteriole penetrating the optic tract. All VP were rich in synaptic contacts. The possibility that the VP of neurosecretory cells may be functionally related to the subarachnoid CSF and the arteriolar blood flow is discussed.
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  • 92
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    Cell & tissue research 236 (1984), S. 561-566 
    ISSN: 1432-0878
    Keywords: Pituitary gland, pars intermedia ; Peptide hormones ; Dopamine ; Corticotropin-releasing hormone ; 6-Hydroxydopamine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary When rats were injected with 6-hydroxydopamine the catecholaminergic nerve terminals in their intermediate lobes exhibited distinct signs of degeneration. Morphometric examination of the Golgi apparatus in cells of the intermediate lobe of these rats showed significant enlargement of Golgi cisternae. The release of adrenocorticotropin, β-endorphin/lipotropin and α-melanotropin from intermediate-lobe cells in vitro was measured by radioimmunoassay. The high basal peptide release was inhibited by dopamine and stimulated by methyl-isobutyl-xanthine. In contrast, γ-aminobutyric acid, serotonin, histamine and noradrenaline, or corticotropin-releasing hormone, rat hypothalamic extract and vasopressin had no or only very weak effects. These observations indicate that the synthetic apparatus of intermediate-lobe cells is constantly depressed by dopaminergic nerves. We were not able to stimulate peptide release from intermediate-lobe cells by use of the abovementioned endogenous agents.
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  • 93
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    Cell & tissue research 237 (1984), S. 169-179 
    ISSN: 1432-0878
    Keywords: Foetal pancreas ; β Cells ; Insulin ; Fasting mothers ; Morphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary After maternal fasting for 72 h the pancreatic β cells of 18-day-old foetal rats show a conspicuous enrichment in secretory material, with an increase of pancreatic insulin concentration and a marked development of the rough endoplasmic reticulum and the Golgi apparatus. The morphometric analysis shows that the intracytoplasmic migration of the secretory granules is inhibited, principally inside the cell web. Consequently the number of secretory granules fused with plasma membrane decreases and this is associated with a decreased foetal plasma insulin. The difference in the ultrastructural aspect of the β cells of foetuses from fasting mothers and of foetuses from fed mothers is less conspicuous at 19 days of gestation and progressively disappears at 20 and 21 days. The modifications in ultrastructural aspect and in functional state are discussed.
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    Cell & tissue research 238 (1984), S. 81-85 
    ISSN: 1432-0878
    Keywords: Retina ; Ibotenic acid ; Toxicity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary After 2 h intraocular injections of 19 and 190 nmoles ibotenic acid in the rat retina produced an intensive vacuolization of the inner plexiform layer and cellular alterations, in the inner nuclear layer and ganglion cell layer. These alterations consisted of either cytoplasmic swelling accompanied by clumping of the nuclear chromatin or darkening of the cytoplasm along with nuclear condensation. A week later the retinas were thinner than the controls due to the disappearance of the affected cells. Pre-treatment with diazepam prevents the morphological alterations induced by 19 nmoles ibotenic acid; mainly the swelling, which was completely prevented, while the darkening was reduced drastically, although some vacuolization of the inner plexiform layer is still present.
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    Cell & tissue research 238 (1984), S. 159-163 
    ISSN: 1432-0878
    Keywords: Ovary ; Rat ; Cell division ; Luteinizing hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of an LH pulse on the rate at which 3H-thymidine is incorporated into cultured ovaries of metestrous rats was studied. In comparison to ovaries cultured with tonic LH, an LH pulse (1) “rescued” follicles from atresia, (2) induced thecal cell proliferation, and (3) increased the rate at which granulosa cells enter mitosis. It is concluded that LH pulses increase follicular growth by first triggering thecal cell proliferation and then inducing mitotic divisions within the granulosa cells of both atretic and non-atretic follicles.
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  • 96
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    Keywords: Choroid plexus ; Immunoglobulin G ; Permeability ; Anti-HRP-IgG ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of autologous antiperoxidase immunoglobulin G (IgG) was studied in the choroid plexus of Lewis rats immunized against horseradish peroxidase (HRP). This experiment was performed to study the permeability of the choroid plexus to intravascular IgG. It was shown that autologous IgG was present in the extravascular spaces. The transendothelial transfer appeared to occur mainly via the fenestrations and some interendothelial junctions. No transfer of IgG at the level of epithelial cells toward the cerebrospinal fluid was demonstrated. Interstitial spaces in contact with the connective-tissue cells of the choroid stroma were strongly labeled. The significance of these spaces remains hypothetical and raises the question of the fate of IgG from the interstitial space.
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  • 97
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    Cell & tissue research 238 (1984), S. 191-195 
    ISSN: 1432-0878
    Keywords: Glial fibrillary acidic protein (GFAP) ; Pituicytes, neonatal ; Development, ontogenetic ; Immunofluorescence ; Organ culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The appearance and intracellular localisation of glial fibrillary acidic protein (GFAP) in pituicytes in neural lobe cultures of newborn rats aged 7 to 30 days were investigated by use of the indirect immunofluorescence method. GFAP-immunoreactive cells were observed mostly in the outgrowth zone. GFAP was localised in the perikaryal cytoplasm as well as in pituicyte processes. GFAP-positive pituicytes showed considerable morphological polymorphism. The presence of GFAP — astrocytic marker — in pituicytes in vitro and the evident morphological similarity to cultured astrocytes suggest the astroglial character of these cells.
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  • 98
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    Keywords: Cyclic guanosine monophosphate-phosphodiesterase (cGMP-PDEase) ; New cytochemical method ; Retina ; Rods, outer segments ; Light perception ; 5′GMPase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cyclic guanosine monophosphate-phosphodiesterase (cGMP-PDEase) activity was studied histo- and cytochemically in the retinal rods of the rat with the use of a newly developed technique. Intense activity of cGMP-PDEase was evenly distributed over the outer segments of the rods. Reaction product was observed on the plasmalemma and on the disk membranes of the outer segments. A weak reaction product occurred also on the plasmalemma of the inner segments; however, no precipitate was found in the perinuclear and synaptic portions of the rod cells. The enzyme activity was strongly inhibited by 2 mM theophilline and by 2 mM 3-isobutyl-1-methylxanthine (IBMX). To confirm the specificity of this new cGMP-PDEase method, the localization of 5′nucleotidase (5′GMPase) was also studied. In contrast to the activity of cGMP-PDEase, the activity of 5′GMPase was distributed on the plasma membrane of the photoreceptor cells extending over a wide range from the synaptic endings in the outer plexiform layer to the tip of the outer segments.
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  • 99
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    Keywords: S-100 ; Müller cell ; Astrocyte ; Development ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The rat retina was studied by immunohistochemistry with antibody to S-100 protein during the first three postnatal weeks. Immunoreactive astrocytes are first detected subjacent to the inner limiting membrane close to the optic disc. They gradually increase in number and spread toward the ora serrata along the inner surface of the retina as the development proceeds. S-100-immunostained Müller cells are first identified on the 12th postnatal day although their immunoreactivity is much weaker than that of astrocytes at the same stage. This differential intensity of the immunoreactivity of the two cell types facilitates observation of the entire shape of the astrocyte. This characteristic reveals that cellular investments of blood vessels in the inner retina are formed by astrocytic processes whereas those in the outer plexiform layer are derived from processes of Müller cells. The cellular investment becomes complete by the 18th postnatal day.
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  • 100
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    Cell & tissue research 238 (1984), S. 559-564 
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Macrophages ; Ovariectomy ; Rat ; Estradiol administration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Macrophages of the adrenal cortex were studied in normal male and female, ovariectomized and estradiol-injected rats. In normal male rats few macrophages with numerous granules were observed in the zona fasciculatazona reticularis border, and in the zona reticularis. Granules, identified as lysosomes, were limited by a single membrane with a heterogeneous matrix; they exhibited acid phosphatase- and aminotriazole-resistant peroxidatic activities. A larger number of macrophages had identical distributions in normal female rats. In ovariectomized and estradiol-injected rats the number and distribution of adrenal macrophages were similar to those in normal females; however, in spayed animals the number of these cells in the zona reticularis was higher than in the other experimental groups. Lysosomes in macrophages of treated animals were more numerous and their contents more complex than in normal male animals. These results indicate that the adrenal macrophage system is stimulated in experimental conditions involving high levels of circulating estrogens.
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