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  • Drosophila  (87)
  • Calcium  (67)
  • Springer  (154)
  • 1995-1999
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  • 1940-1944
  • 1925-1929
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  • 1980  (43)
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  • 1995-1999
  • 1990-1994  (70)
  • 1980-1984  (84)
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  • 1925-1929
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  • 1
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    Journal of solution chemistry 21 (1992), S. 525-532 
    ISSN: 1572-8927
    Keywords: Calcium ; molybdate ; aqueous ; thermodynamic ; complexation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The solubility of powellite [CaMoO4(c)] was studied in aqueous Na2MoO4, CaCl2 and Ca(NO3)2 solutions ranging in concentrations from 1×10−4M to 1.0M and over equilibration times extending to 36 days. Our experimental data were interpreted using the aqueous ion-interaction model of Pitzer and coworkers. The Ca2+−MoO 4 2− ion-interactions were found to be analogous to Ca2+−SO 4 2− . The use of Ca2+−MoO 4 2− ion-interactions parameters (β(0)=0.2, β(1) = 3.1973 and β(2)) and a logK sp of −7.93 gave excellent predictions of all of the experimental data. Commonion ternary interaction parameters such as MoO 4 2− −Cl− or MoO 4 2− −NO 3 − were not required.
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  • 2
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    Calcified tissue international 30 (1980), S. 209-216 
    ISSN: 1432-0827
    Keywords: Calvarium ; cAMP ; Vitamin D3 metabolites ; Calcium ; Parathyroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Results from in vitro works suggest that 1,25- and 24,25-dihydroxyvitamin D3 (1,25-(OH)2D3 and 24,25-(OH)2D3) act on bone via different mechanisms. The present investigation was performed to study the effect of these two metabolites and of their precursor 25-hyxdroxyvitamin D3 (25-(OH)D3) on bone cAMP content in vitro. Rats' paired half calvaria were incubated under sterile conditions with one vitamin D3 derivative (10−13 to 10−9 M) or with ethanol (0.005 ml for 15 min to 24 h in 1 ml medium containing 0, 0.2, 1, 2, or 3 mM calcium. In some experiments: (a) cycloheximide (10−5M) was added simultaneously with the vitamin D3 metabolites; (b) 1–84 bPTH (5 × 10−8 M) was added for 5 or 15 min at the end of the 24 h incubation. Calvaria were immersed in 1 ml TCA 5% 4°C and homogenized. The cAMP was extracted with diethylether and measured by a competitive protein binding assay. Results bring further evidence for a particular effect of low doses of 24,25-(OH)2D3 (10−9 to 10−12M) and of 25-(OH)D3 (10−9 to 10−11M) on bone, different from that of 1,25-(OH)2D3: cAMP content was higher in 24,25-(OH)2D3- or 25-(OH)D3-treated and lower in 1,25-(OH)2D3-treated calvaria than in ethanol-treated ones with 1 mM calcium. The 1,25-(OH)2D3 effect persisted in calcium-free medium whereas 25-(OH)D3 and 24,25-(OH)2D3 effects could not be observed with 0 mM nor with 3 mM calcium. The required duration of the preincubation (over 1 h) as well as the inhibitory action of cycloheximide may suggest an involvement of protein synthesis in the vitamin D3 metabolites effects. Neither 1,25-(OH)2D3 nor 24,25-(OH)2D3 affected the PTH-induced increase in bone cAMP content.
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  • 3
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    Calcified tissue international 32 (1980), S. 201-206 
    ISSN: 1432-0827
    Keywords: Magnesium ; Parathyroid hormone ; Secretion ; In vitro ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In a well-defined in vitro perifusion system, the effects of extracellular magnesium concentration (Mg) on parathyroid hormone (PTH) secretion by bovine parathyroid tissue were examined. At Mg less than 0.8 mM, the ability of the glands to secrete hormone maximally in response to low calcium (Ca) stimulation was progressively impaired. Low Mg also impaired the ability of isoproterenol, dibutyryl cyclic AMP and theophylline to stimulate hormone release. The defect in hormone release at low Mg observed in vitro was analogous to the well-documented inhibition of secretion observed in vivo. Increases in Mg from 0 to 0.8 mM rapidly repaired the defect in hormone secretion. At Mg above 1.0 mM there was a Ca-like effect on hormone release, with a progressive decrease in secretion at increased Mg. Although its mechanism is not yet clear, the low Mg effect appears to impair principally the process of hormone release rather than its biosynthesis or storage.
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  • 4
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    Calcified tissue international 50 (1992), S. 391-396 
    ISSN: 1432-0827
    Keywords: Bone ; Matrix ; Phosphoproteins ; Biomineralization ; Calcium ; Nucleation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Osteopontin, bone sialoprotein, and bone acidic glycoprotein-75 are three acidic phosphoproteins that are isolated from the mineralized phase of bone matrix, are synthesized by osteoblastic cells, and are generally restricted in their distribution to calcified tissues. Although each is a distinct gene product, these proteins share aspartic/glutamic acid contents of 30–36% and each contains multiple phosphoryl and sialyl groups. These properties, plus a strict relationship of acidic macromolecules with cell-controlled mineralization throughout nature, suggest functions in calcium binding and nucleation of calcium hydroxyapatite crystal formation. However, direct proof for such roles is still largely indirect in nature. The purpose of this review is to present two speculative hypotheses regarding acidic phosphoprotein function. The goal was to use new sequence information along with database comparisons to develop a structural rationalization of how these proteins may function in calcium handling by bone. For example, our analysis has identified a conserved polyacidic stretch in all three phosphoproteins which we propose mediates metal binding. Also, conserved motifs were identified that are analogous with those for casein kinase II phosphorylation sites and whose number correlates well with that of phosphoryl groups/protein. A two-state conformational model of calcium binding by bone matrix acidic phosphoproteins is described which incorporates these findings.
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  • 5
    ISSN: 1432-0827
    Keywords: Calcitonin ; Antibodies ; Calcium ; Osteoporosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In order to evaluate the potential inhibition of the acute anti-osteoclastic activity of salmon calcitonin (SCT) by specific antibodies (Ab), we compared the SCT-induced hypocalcemic effect in young male rabbits with significant titers of high affinity Ab and in matched animals without Ab. Immunization of rabbits was performed by repetitive s.c. injections of SCT and Freund adjuvant. Ab were present in four-fifths of SCT-treated rabbits (Ab+). Their titer varied from 0.8×10(-9) to 30×10(-9) M/liter and their constant of affinity from 0.97×10(9) to 4.2×10(9) L/M. Intravenous injection of 1 IU/kg SCT to Ab+ rabbits induced a significant decrease (P〈0.01) of ionized serum calcium (Ca2+) after 30 minutes (mean±SD:-9±0.6%) and until the 240th minute of the test (-16.7±4.7%), with a maximum after 120 minutes (-22.6±2%). This was not significantly different from the hypocalcemic effect measured after the same procedure performed in matched animals without Ab (Ab-): significant decrease in Ca2+ (P〈0.01) after 30 minutes (-8.2±2.2%), maximal after 150 minutes (-23.2±4.9%), and lasting until 210 minutes (-14.5±3.7%). We conclude that, in the particular model of the male young rabbit, specific anti-SCT Ab do not block or reduce the acute anti-osteoclastic activity of SCT.
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  • 6
    ISSN: 1432-0827
    Keywords: Computed tomography ; Trabecular bone density ; Strontium ; Calcium ; Atomic absorption spectroscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Changes in the average linear attenuation coefficient (LAC) within a fixed measurement volume in the proximal end of the dog tibia, which contains trabecular bone and associated soft tissues (the trabecular bone “space”), were monitored continuously using gamma-ray computed tomography (γ-CT) prior to, during, and following intravenous infusion of strontium (Sr) lactate. An infusion of 1.3–4.7 g of Sr over a period of 110–160 minutes into 20-kg dogs resulted, within 6–8 hours, in an increase of 0.019–0.045 cm-1 (P〈0.002) in the LAC. Calibration of the γ-CT system showed that 0.44 mg/cm3 of Sr produced a change of 0.01 cm-1 in the LAC. Using this conversion factor, the Sr concentration in the trabecular bone space resulting from infusion, as measured by flame atomic absorption spectroscopy, agreed with that predicted by the change observed in the LAC. Sr present in the serum and urine was consistent with the changes observed in the LAC over the study period. Control dogs infused with mineral-free solutions showed no change in LAC. Calcium equivalents required to give the changes observed in the LAC using Sr indicate that variations in skeletal turnover in man can be monitored in the peripheral skeleton using γ-CT.
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  • 7
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    Calcified tissue international 51 (1992), S. 42-47 
    ISSN: 1432-0827
    Keywords: Kidney ; Endoplasmic reticulum ; Calcium ; In vivo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Calcium uptake by the endoplasmic reticulum (ER) is important for cellular calcium homeostasis, yet its regulation in nonmuscle cells is poorly understood. We reported that Ca2+ uptake by a light fraction of canine renal cortical ER (LER) is stimulated by protein kinase C in vitro. Here we describe conditions in vivo that stimulated renal cortical LER Ca2+ uptake. Thirty minutes after contralateral nephrectomy in the dog, 45Ca2+ uptake into renal cortical LER was increased 42% above control LER. There was no difference in LER Ca2+ uptake 24 hours after uninephrectomy. Acute denervation did not reproduce the increase in LER 45Ca2+ uptake seen at 30 minutes after uninephrectomy, nor did prior thyroparathyroidectomy abolish it. Forty-eight hours after thyroparathyroidectomy, 45Ca2+ uptake activity into renal cortical LER was decreased ≈sevenfold. In a proximal tubular cell line (LLC-PK1), 30-minute incubation with 12-O-tetradecanoylphorbol-13-acetate doubled 45Ca2+ uptake into a nonmitochondrial pool. Pretreatment with epidermal growth factor halved ER Ca2+ uptake, whereas insulin-like growth factor and growth hormone, alone or in combination, had no effect. Our data suggest that Ca2+ uptake into renal cortical ER is stimulated acutely during compensatory renal growth, perhaps through protein kinase C, and is stimulated chronically by parathyroid hormone.
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  • 8
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    Journal of molecular evolution 35 (1992), S. 51-59 
    ISSN: 1432-1432
    Keywords: Drosophila ; Gart locus ; Chironomus tentans ; Purine nucleotide biosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Drosophila Gart locus consists of two genes. One gene encodes three enzymes in the de novo purine nucleotide biosynthesis pathway [glycinamide ribonucleotide synthetase (GARS), aminoimidazole ribonucleotide synthetase (AIRS), and glycinamide ribonucleotide transformylase (GART)]. The second gene lies within an intron of the purine gene and encodes a cuticle protein. To investigate the evolution of the Gart locus, the Chironomus tentans homolog was cloned by screening a genomic DNA library with a polymerase chain reaction product. This study shows that the interesting structural features of this locus conserved in two distant Drosophila species are not found in the Chironomus homolog. These features include the cuticle protein gene nested within an intron and the existence of an alternative transcript to yield a monofunctional enzyme. In addition, the extremely rapid divergence of coding sequence seen for members of the tandemly duplicated AIRS domain in Drosophila is found to be much less rapid in Chironomus.
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  • 9
    ISSN: 1432-1432
    Keywords: Drosophila ; per gene ; Threonine-Glycine ; repeat sequence ; melanogaster subgroup phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Threonine-Glycine (Thr-Gly) region of the period gene (per) in Drosophila was compared in the eight species of the D. melanogaster subgroup. This region can be divided into a diverged variable-length segment which is flanked by more conserved sequences. The number of amino acids encoded in the variable-length region ranges from 40 in D. teissieri to 69 in D. mauritiana. This is similar to the range found within natural populations of D. melanogaster. It was possible to derive a Thr-Gly “allele” of one species from that of another by invoking hypothetical Thr-Gly intermediates. A phylogeny based on the more conserved flanking sequences was produced. The results highlighted some of the problems which are encountered when highly polymorphic genes are used to infer phylogenies of closely related species.
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  • 10
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    Journal of molecular evolution 34 (1992), S. 130-140 
    ISSN: 1432-1432
    Keywords: Drosophila ; Sophophora ; cDNA-DNA hybridization ; Phylogenetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have performed DNA-DNA hybridization experiments among several species of Drosophila using the evolutionarily conserved portion of the genome representing sequences coding for amino acids of proteins. This was done by using as tracer, radioactively labeled complementary DNA that was reverse transcribed from adult mRNA. We show that this procedure extends phylogenetically the distance over which the technique can be applied to fast-evolving groups such as Drosophila. The major phylogenetic conclusions are (1) the subgenus Sophophora is a monophyletic lineage; (2) within Sophophora the melanogaster subgroup is closer to the obscura group than either group is to the willistoni group; (3) the subgenus Drosophila is complex with most major lineages originating deep in the phylogeny; the subgenus may not be monophyletic; (4) as with most groups classically placed in Drosophila, the Hawaiian Drosophila originate early, supporting the notion that this lineage is older than the extant islands; and (5) the virilis/repleta lineage is monophyletic within Drosophila.
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  • 11
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    Journal of comparative physiology 170 (1992), S. 311-316 
    ISSN: 1432-1351
    Keywords: Phototransduction ; Limulus polyphemus ; Inositol trisphosphate ; Calcium ; Latency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The latent period before depolarization of Limulus ventral photoreceptors by light flashes was compared with that following brief, intracellular, pressure-injection of d-myo-inositol 1,4,5 trisphosphate. At temperatures between 18 °C and 22 °C and with an extracellular calcium concentration of 10 mM, the responses of 4 cells to light and to injections of 100 μM inositol trisphosphate displayed average latencies of 71 and 56 ms, respectively. The latencies of responses to InsP3 included an estimated 20 ms dead-time inherent in the injection method. Reducing the temperature lengthened the latency of the response to light (Q10 approximately 3.2 between 7 and 22 °C) more than that to inositol trisphosphate (Q10 approximately 2.3). Bathing the photoreceptors in seawater containing no added calcium and 1 mM of the calcium chelator EGTA greatly increased the latency of the light response at all temperatures, but did not increase the latency of the response to inositol trisphosphate. We conclude that the response to inositol trisphosphate lacks the calcium- and temperature-sensitive latent period which characterizes the response to light. If inositol trisphosphate acts, via the release of stored calcium, to stimulate an intermediate in the visual cascade, then that intermediate would appear to be downstream from the latency-generating mechanism.
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  • 12
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    Journal of comparative physiology 171 (1992), S. 413-419 
    ISSN: 1432-1351
    Keywords: Calcium ; Cobalt ; Lateral line ; Behavior
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hypothesis that the blind cave fish (Astyanax hubbsi) adjusts the level of stimulation to its lateral line system (LLS) by varying its own velocity was examined. When the sensitivity of the LLS sense organs was reduced by lowering the Ca2+ concentration in the water or by adding Co2+ the fish compensated for this by swimming at a higher velocity.
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  • 13
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    Journal of molecular evolution 16 (1980), S. 37-46 
    ISSN: 1432-1432
    Keywords: Evolution ; Drosophila ; Temperature ; Mitochondrial enzymes ; Kinetic properties
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The evolutionary behavior of two mitochondrial enzymes (L-glycerol 3-phosphate:cytochrome c oxidoreductase E.C.1.1.1.95,αGPO, and L-malate: NAD+ oxidoreductase, E.C.1.1.1.37, m-MDH) obtained from several temperate and tropicalDrosophila species was examined by comparing their catalytic properties, which related to temperature (Km-Ea-Q10-Thermostability). MitochondrialαGPO or m-MDH obtained either from temperate or from tropical species was found to exhibit similar catalytic properties while for both cytosolic enzymes, theαGPDH and s-MDH, Km patterns were similar among species from the same thermal habitat and different between thermal habitats. In combination with other observations reported in the literature these facts support the view that the function, and probably the structure, of mitochondrial enzymes are better conserved in evolution than those of the corresponding enzymes found in the cytosol. It is proposed that the relative invariance of the mitochondrial enzymes structure is probably linked to a necessary relative invariance of molecular interactions inside the mitochondrion.
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  • 14
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    Development genes and evolution 188 (1980), S. 55-63 
    ISSN: 1432-041X
    Keywords: Drosophila ; Compound eye ; shibire ts ; Development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analysed the effect of temperature on both developing and adult eye cell clones homozygous forshi ST139, a temperature-sensitive mutant ofDrosophila melanogaster. The mutant gene, autonomous in its cellular expression, causes structural modifications of ommatidial cells when adult clones of cells are exposed to the restrictive temperature (29°C) for several days. However, the mutant phenotype reverses to normal within 4 days at the permissive temperature (20°C). The results of pulse, shift-up and shift-down experiments show that the temperaturesensitive period for developing compound eye cells is from the late second instar up to the early pupa. Cytodifferentiation of compound eye cells is blocked by restrictive temperature treatment during this period, whereas cell proliferation does not seem to be directly affected. These results are discussed with regard to the other known aspects of the phenotype observed in mutant individuals.
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  • 15
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    Development genes and evolution 192 (1983), S. 164-170 
    ISSN: 1432-041X
    Keywords: Drosophila ; Imaginal disc ; Morphogenesis ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The early morphogenesis of the eye-antennal disc ofDrosophila in response to 20-hydroxy ecdysone involves the curling of the eye anlagen dorsally over the antenna. During this process, the area of the peripodial membrane is substantially reduced. The peripodial membrane is taut at this stage, and if it is cut the curling of the disc cannot continue, and the eye anlagen returns to its original position within one minute of the operation. In contrast, cutting the columnar epithelium between the eye and antennal anlagen does not disrupt curling, but actually facilitates it. During curling, the cells of the peripodial membrane appear healthy, and exhibit basal extensions. We suggest that the curling of the eye is mediated by the conversion of cuboidal peripodial membrane cells into pseudostratified columnar epithelium at the edges of the peripodial membrane. Subsequently, cells of the peripodial membrane secrete first a pupal cuticle, and then an imaginal cuticle.
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  • 16
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    Development genes and evolution 192 (1983), S. 275-279 
    ISSN: 1432-041X
    Keywords: Evagination ; Morphogenesis ; Metamorphosis ; Female genital disc ; Drosophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The morphology of the evaginating female genital disc ofDrosophila melanogaster was examined at different stages of metamorphosis. The observations show that the internal genital organs are derived from the anterior half of the disc and that their morphogenesis is mainly a protrusion of the different primordial areas of the disc epithelium. The external genital and anal derivatives originate from the posterior half of the disc, which undergoes complex rearrangements during metamorphosis. The disc opens along the posterior margin and the dorsal and ventral epithelia evert and thereby completely reverse their anteroposterior orientation. Dramatic elongation has been observed during the formation of the seminal receptacle. The cells of the repressed male genital primordium do not form any recognizable structures and are assumed to be eliminated during metamorphosis.
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  • 17
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    Development genes and evolution 192 (1983), S. 299-302 
    ISSN: 1432-041X
    Keywords: Differentiation ; Teratogens ; Drosophila ; 5-Azacytidine ; Methylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effects of cytidine and cytidine analogs were studied inDrosophila embryonic cell cultures and two wild-type established cell lines, Oregon-R and Schneider line 2. Primary embryonic cultures have been shown to be an excellent system for the study of embryonic development; a number of cell types undergo normal differentiation in vitro. Treatment of these cultures with putative teratogens resulted in an inhibition of muscle and/or neuron differentiation in our study. Treatment of these cells with cytidine and seven other analogs had no effect on neuron and muscle differentiation. The compound 5-azacytidine, when added to primary cell cultures, inhibited normal differentiation at subtoxic doses while inducing the production of three proteins that comigrate with the heat-shock proteins, hsp 23, 22a and 22b. 5-Azacytidine did not stimulate differentiation in Oregon-R or SchneiderDrosophila cell lines. The in vitro blockage of differentiation by 5-azacytidine suggests that it may act as a teratogen.
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  • 18
    ISSN: 1420-9071
    Keywords: Aromatic plants ; essential oils ; Drosophila ; insecticides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Effects of the essential oils (EOs) extracted from eleven aromatic plants belonging to the Lamiaceae family (common in the Greek flora) were examined upon three different developmental stages ofDrosophila auraria. All of the EOs examined exhibited insecticidal effects, either by preventing egg hatching, or by causing the death of larvae and adult flies. In several cases, malformation and/or prohibition of puparium formation was also observed.
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  • 19
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    Development genes and evolution 192 (1983), S. 280-284 
    ISSN: 1432-041X
    Keywords: Evagination ; Morphogenesis ; Metamorphosis ; Intersexual genital disc ; Drosophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Morphogenetic movements of the intersexual genital disc of thedoublesex-dominant mutant ofDrosophila melanogaster were followed during metamorphosis. Intersexual genital discs contain well developed genital primordia of both sexes as well as an anal primordium, and all of these primordia evaginate simultaneously. The female genital primordium is deflected to the ventral side by the male genital primordium which is located anterior to it. Subsequently the anterior parts of the two genital primordia project their internal appendages in parallel in the anterior direction. The morphogenetic movements closely resemble those of the corresponding parts of normal males and females. The disc opens at the stalk along the posterior edge and the two genital primordia completely evert their posterior parts. These areas undergo complex rearrangements whereby the anlage for the male genital arch as well as that for the 8th tergite evert and move around the lateral side of the disc. They both fuse dorsally after enclosing the anal tube. The formation of the characteristic abnormalities of the intersexual genitalia seems not to result simply from spatial problems of the simultaneous evagination of the genital anlagen but rather to be a direct result of the ambiguous genetic signalling in the intersexual cells of these primordia.
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  • 20
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    Development genes and evolution 192 (1983), S. 337-346 
    ISSN: 1432-041X
    Keywords: Drosophila ; Gynandromorphs ; Genital disc ; Compartments ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genital imaginal disc ofDrosophila differentiates the terminalia, i.e. the genitalia and analia, of both sexes. It represents a composite anlage, containing a female genital primordium, a male genital primordium and an anal primordium. In normal males and females, only one of the two genital primordia differentiates; the other is developmentally repressed. Therefore, cell-lineage relationships between the male and female genital primordia can only be studied in sexual mosaics which differentiate female and male cells. We producedMinute (M)‖non-Minute(M+) gynandromorphs and selected those with sexually mosaic terminalia for a cell-lineage analysis. In these mosaics, either the male (XO) or female (XX) cells wereM + and thus had a growth advantage. The differential growth rates served as a tool to detect clonal restrictions. In control gynandromorphs (M +‖M +), the amount of female genitalia differentiated was largely independent of the amount of male genitalia present. In contrast, male and female anal structures, as a rule, added up to one full set. The same was true for the experimentalM‖M + gynandromorphs, but the contribution ofXX andXO cells to mosaic terminalia changed drastically due toM + cells competing successfully against the more slowly growingM cells. Specific subsamples ofM‖M + gynandromorphs showed thatM cells in a non-mosaic primordium are shielded from cell competition taking place in the neighbouring mosaic primordium. We conclude that the three primordia of the genital disc represent developmental compartments. In the genital primordia, even developmentally repressedM + cells compete successfully against developmentally activeM cells.
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  • 21
    ISSN: 1432-041X
    Keywords: Muscle ; Salivary glands ; Gut ; Programmed cell death ; Steroid hormones ; Drosophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In holometabolous insects, the steroid molting hormone 20-OH-ecdysone (ecdysterone) orchestrates the diverse developmental events of metamorphosis, in large part by regulating gene expression. In Drosophila, the Broad Complex (BR-C) is one of the first loci to be induced by ecdysterone at the end of larval life, and is essential for translating the hormonal signal into the behavioral and anatomical events which herald the onset of metamorphosis. BR-C products are believed to act by binding to and modifying the transcriptional activities of other hormone-sensitive genes. In addition to abnormalities of the epidermis, BR-C mutants dying during metamorphosis manifest a syndrome of multiple internal tissue defects which represent a failure of the larval-to-adult transition. We have reported features of central nervous system metamorphosis requiring BR-C function, notably morphogenetic movements and optic lobe organization. In this paper we describe defective development of salivary glands, flight muscles, and gut in BR-C mutants, including: persistence of larval salivary glands; failure of the adult salivary glands to extend into the thorax; abnormalities of midgut transition and of proventriculus structure and location; and absence of dorsal-ventral indirect flight muscles. Some of these abnormalities represent defects in programmed cell death. Distinct patterns of phenotypes were seen in mutants of each of the three lethal complementation groups comprising the BR-C. The patterns of phenotypes suggest overlapping but distinct functions encoded by this complex locus.
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  • 22
    ISSN: 1432-041X
    Keywords: Drosophila ; Imaginal discs ; Pattern formation ; rotund
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary InDrosophila imaginal discs, pattern formation requires the activity of three positional information systems, antero-posterior (A/P), dorso-ventral (D/V) and proximo-distal (P/D). Three genes,Decapentaplegic, Distal-less androtund (rn), involved in pattern formation along the P/D axis have been characterized. Thern gene is required in a sub-distal region, localized at a similar position along the P/D axis in all appendages; it encodes two major transcripts, m1.7 and m5.3, both expressed in the central region of all the major imaginal discs. The present study of these transcripts in severalrn mutant favours m5.3 as encodingrn morphogenetic function in the imaginal discs. The fine characterization of its distribution partitions all major imaginal discs in domains along the P/D axis. The ventral and dorsal discs appear to be similarly but not identically organized: two P/D domains are evident in the wing and haltere discs whilst the leg and antenna discs appear to be composed of at least three. We also show that m5.3 is sex-regulated in the genital disc and thatrn function is required for proper development of a sub-distal structure of the female genitalia. This suggests that the primordia of the female genitalia may be organized in a similar way to the other imaginal discs, and strongly supports the hypothesis thatrn function is specific to pattern formation along the P/D axis and that it may be involved in the establishment or maintenance of this pattern.
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  • 23
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    Development genes and evolution 201 (1992), S. 364-375 
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    Keywords: Drosophila ; Tissue culture ; In vitro ; Invertebrate embryogenesis
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    Notes: Summary We have devised techniques to culture whole, dissected embryos of Drosophila melanogaster. We examine multiple aspects of the morphological and physiological development of the epidermis, musculature, nervous system, and internal organs in this cultured preparation, and show that in vitro development closely parallels normal embryogenesis. These techniques permit a wide range of experimental manipulations during embryogenesis and allow us to extend observations through late embryonic stages, after cuticle deposition. Applications of this technique are presented.
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  • 24
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    Development genes and evolution 188 (1980), S. 163-177 
    ISSN: 1432-041X
    Keywords: Yolk sac ; Ultrastructure ; Embryogenesis ; Drosophila
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    Topics: Biology
    Notes: Summary Changes at the ultrastructural level during germ band extension in the embryo ofDrosophila melanogaster are described. Cytoplasmic connections between cells and the yolk sac are present during initial cellular movements. At this time, a continuous system of microfilaments is present adjacent to the membranes in the connections and at the periphery of the yolk sac. As germ band extension progresses, this system becomes discontinuous, and microfilaments are apparent only in the immediate vicinity of the connections. Cytoplasmic connections are disassembled at approximately the midpoint of extension; at the same time, extensive membrane associations develop between germ band cells and between these cells and adjacent yolk sac membranes. Positioning and orientation of cytoplasmic connections suggest that the yolk sac, via these connections, is actively involved in the cellular movements of early germ band extension.
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  • 25
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    Development genes and evolution 189 (1980), S. 57-67 
    ISSN: 1432-041X
    Keywords: Drosophila ; Ecdysone deficient mutants ; Ecdysteroid titer ; Ring gland ; Fine structure
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    Notes: Summary This paper describes two ecdysone-deficient, recessive-lethal mutants,lethal(1)giant ring gland (grg) andlethal(1)suppressor of forked mad-ts (mad-ts: Jürgens and Gateff 1979) and compares their ecdysteroid titers with that of the wild-type. Mutant larvae show a much reduced ecdysteroid content, amounting to 1/10 to 1/30 of the wild-type values, but never a true titer peak. They fail to pupate and die after 1–3 weeks. Ecdysteroid feeding elicits different responses in the larvae of the two mutants.mad-ts larvae pupate within 24 h, thus showing that their low ecdysteroid titer is directly connected to their inability to pupate.mad-ts resembles the mutantlethal (3)ecdysone-1 ts (Garen et al. 1977). Thegrg mutant larvae, on the other hand, fail to pupate after 20-hydroxyecdysone feeding as well as injection. The primary defect of thegrg mutant is not entirely clear. Thegrg larval salivary gland cells appear to possess normal ecdysteroid receptors. Furthermore, the low ecdysteroid titer ingrg is not the result of an increased ecdysteroid catabolism. The primary defect in the mutant may lie in the malfunctioning neurosecretory cells which do not show neurosecretion in histological preparations. Further support for this notion comes from electronmicrographs of the enlargedgrg ring glands which, in contrast to the wild-type, do not possess nerve endings. In the wild-type three ecdysteroid peaks were found: one shortly before puparium formation, the second at approximately 12 h and the third at about 30 h after pupation. The ecdysteroid titer peak in late third instar, wild-type larvae is mainly due to the presence of 20-dydroxyecdysone as shown by radioimmunoassays after thin layer chromatography and derivatization followed by gas liquid chromatography and mass spectroscopy. In addition, a number of unidentified polar and apolar metabolites were also present.
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    Development genes and evolution 188 (1980), S. 157-161 
    ISSN: 1432-041X
    Keywords: Drosophila ; Imaginal discs ; Compartments ; Distal outgrowth
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    Topics: Biology
    Notes: Summary Peripheral tissue of the imaginal wing disc gives rise to the proximal mesothoracic structures of the adult. Pieces of peripheral tissue, which have no regenerative capacity when cultured as intact fragments, are capable of distal outgrowth (regeneration) after dissociation and reaggregation. This ability depends on the region of the disc periphery from which the fragment is taken. Extensive distal outgrowth occurs in reaggreages of a fragment containing equal proportions of tissue from anterior and posterior developmental compartments. The extent of outgrowth decreases as the proportion of posterior tissue is reduced, so that a fragment containing only anterior tissue shows no regeneration after dissociation. Limited distal outgrowth occurs in reaggregates of a wholly posterior fragment, but the regenerative capacity is increased greatly when a small amount of anterior tissue is included. It is concluded that distal outgrowth in the wing disc requires an interaction between cells of the anterior and posterior compartments.
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  • 27
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    Development genes and evolution 189 (1980), S. 91-96 
    ISSN: 1432-041X
    Keywords: Epimorphic regulation ; Drosophila ; Imaginal discs
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    Topics: Biology
    Notes: Summary It has been known for many years that when a wing disc ofDrosophila is bisected, and the fragments cultured in adult females, regulation occurs and either a complete disc is regenerated or the fragment is duplicated. We have investigated how this regeneration process occurs. To establish which cells contribute to the regenerate, and thus determine if regeneration is the result of epimorphic regulation, fragments of discs, after culture in an adult for one to five days, were exposed to3H-thymidine to label replicating cells. Imaginal discs, both whole and as regenerating fragments, undergo some DNA replication which is distributed throughout the disc, but cut discs frequently show clusters of labelled cells around the wound, indicating that regeneration is probably epimorphic.
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  • 28
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    Development genes and evolution 192 (1983), S. 48-50 
    ISSN: 1432-041X
    Keywords: Drosophila ; Hybrid lethality ; Imaginal discs ; Interspecific transplantation
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    Notes: Summary Females ofDrosophila melanogaster, crossed with males ofDrosophila mauritiana, produce only female offspring. The male hybrid larvae grow very slowly, fail to pupate and die after prolonged larval life. Imaginal discs from these male hybrids transplanted into Drosophila melanogaster larvae can give rise to adult structures with normal patterns. Differentiation of hybrid imaginal disc tissue is improved by short term culture in non-hybrid larvae prior to metamorphosis, suggesting that the hybrid larval haemolymph is inadequate to sustain normal imaginal disc growth. This may represent the physiological basis of the reproductive isolating mechanism separating the twoDrosophila species
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    Development genes and evolution 192 (1983), S. 270-274 
    ISSN: 1432-041X
    Keywords: Fate map ; Repressed primordium ; Sex determination ; Genital disc ; Drosophila
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    Notes: Summary The female genital disc ofDrosophila melanogaster was cut into distinct fragments, and the prospective fates of the fragments were determined by putting them through metamorphosis in host larvae. The dorsal epithelium contains the anlagen for the anal plates and parovaria, as well as the repressed male genital primordium. The ventral epithelium gives rise to all of the female genital structures except for the parovaria. The results were compared with published fate maps and observations made in experiments with sex-transforming mutations. This allowed us to establish a detailed three-dimensional fate map of the female genital disc, which shows a well-developed female genital primordium in the ventral epithelium, a repressed male genital primordium in the anterior part of the dorsal epithelium and an anal primordium in the posterior region of the dorsal disc epithelium.
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  • 30
    ISSN: 1432-041X
    Keywords: Drosophila ; Cell degeneration ; Imaginal disc ; Basal lamina ; Blood cells
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    Notes: Summary The mutationsvestigial (vg; recessive) andUltravestigial (vg U; dominant) ofDrosophila melanogaster give rise to identical mutant adult phenotypes in which much of the cases this results from cell death in the presumptive wing margin of the wing disc in the third larval instar, but the process of cell degeneration is quite different in the two mutants. Invg cell death occurs continuously throughout the third larval instar, while invg U it occurs only in the early third instar. Cells fragment and some of the fragments condense, becoming electron dense (“apoptosis”). Both condensed and ultrastructurally normal cell fragments are extruded to the basal side of thevg disc epithelium. They accumulate under the basal lamina in the wing pouch area until they are phagocytosed by blood cells entering the wing pouch during the six hours following pupariation. Fragments are not extruded from thevg U epithelium but are apparently phagocytosed by neighboring epithelial cells. The basal lamina undergoes mophological changes following pupariation and is phagocytosed by blood cells in both wild-type andvestigial, but investigial the degenerated cell fragments are also engulfed by the same blood cells.
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  • 31
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    Development genes and evolution 192 (1983), S. 317-326 
    ISSN: 1432-041X
    Keywords: Neurogenic mutations ; Topological specificity ; Drosophila
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    Notes: Summary Neurogenic mutations have been found to cause the neuralization of certain regions of the ectoderm and yet to permit normal development of the remaining embryonic cells. Thus, it seems that the activity of the wild-type alleles of these genes is dispensable in a considerable fraction of the embryo during wild-type development. This effect might be a consequence of the cells' position within the embryo; alternatively, it might be independent of the position but be due rather to the genetic activity experienced by the cells previous to their commitment. The results described in this paper indicate that genes controlling patterning along the embryonic dorso-ventral perimeter (dorsal and Toll) are epistatic to genes controlling neurogenesis, their activity deciding which ectodermal cells are susceptible to neurogenesis. Using alleles with low expressivity, evidence was obtained showing that the tracheal placodes define the boundary of the territory which has neurogenic abilities at thoracic and abdominal levels.
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  • 32
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    Development genes and evolution 201 (1992), S. 105-112 
    ISSN: 1432-041X
    Keywords: Drosophila ; Genital disc ; tra-2 ts ; Differentiation
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    Notes: Summary Diplo-X flies homozygous for the transform-er-2 ts (tra-2 ts) mutation develop into females at 16° C, while they develop into males at 29° C (Belote and Baker 1982). By means of this conditional mutation, we have carried out a detailed analysis of the development of the genital disc. Temperature shifts between 16 and 29° C, in both directions, and temperature pulses at 29° C, have been applied during the larval growth of tra-2 ts homozygous diplo-X flies, and the external derivatives of the genital disc have been analysed. Genital discs shifted from 16 to 29° C rapidly lose their capacity to differentiate female genital structures, while they become able to differentiate male genital structures whose inventory is more complete the earlier in larval development the temperature shift is carried out; moreover, duplicated male genital structures were observed. In the shift from 29 to 16° C, the genital disc loses its capacity to differentiate male genital structures, while it becomes able to differentiate female genital structures. The inventory of male structures is smaller, and the inventory of the female structures is more complete, the earlier in larval development the temperature is shifted. No duplicated female or male genital structures were observed in the downshift experiment. With respect to the analia, the shift from 16 to 29° C resulted in the quick formation of pure male anal plates, while in the opposite shift the formation of pure female anal plates occurred gradually. Moreover, the time course for the dorsal and ventral anal plates to show normal female phenotype was different: when the dorsal anal plates were completely normal, it was still possible to find incomplete ventral anal plates. In the pulse experiment at 29° C, the genital disc is able to differentiate both female and male genital structures, although the inventory of the latter ones was not complete. In addition, the capacity of the genital disc to differentiate male genital structures depended on the duration of the temperature pulse. The anal plates were always female, although they showed a reduction in their size, the ventral female anal plate being more affected than the dorsal one. No male anal plates were observed. The results have revealed that the genital disc follows a sequence in its capacity to differentiate female or male adult structures. We suggest that this sequence reflects the sequence of determination events occurring in the genital disc during its larval growth. In addition, results shown here provide evidence for the existence in the female genital primordium of a set of cells capable of giving rise either to female genital structures (ventral vaginal plates) or to male genital structures (hypandrium and penis apparatus). We also present evidence supporting the previous idea of two primordia for the anal plates.
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  • 33
    ISSN: 1432-041X
    Keywords: Drosophila ; Homeotic gene regulation ; Antennapedia ; Development ; β-galactosidase
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    Notes: Summary In order to study the regulation of spatial and temporal expression of the homeotic gene Antennapedia (Antp) in Drosophila melanogaster, we have constructed fusion genes which contain Antp sequences linked to the reporter gene lac Z of Escherichia coli. In one case of P-element transformation, a fusion gene construct integrated into the endogenous Antp gene close to one of the two promoters (P1). The spatial expression from the reporter gene in this transformant line, as analysed by the detection of β-galactosidase activity, was found to exactly mimic the normal expression from the P1 promoter of the Antp gene. We have used this unique transformant as a tool for studying the expression of the P1 promoter in embryonic, larval and adult development. Parallel lines transformed with the same fusion gene construct did not confer a correct P1 pattern of expression. The position in the genome was, therefore, crucial for the expression pattern of the reporter gene. Experiments aiming at the detection of autoregulatory control of Antp gene expression were designed. The results did not, however, support models of positive or negative autoregulation of P1 expression by Amp protein.
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    Development genes and evolution 201 (1992), S. 120-123 
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    Keywords: Integrin ; Drosophila ; In vitro ; Imaginal disc
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    Notes: Summary Drosophila imaginal disc cell lines show a characteristic pattern of aggregation in culture, which appears to be due to cell-cell rather than cell-substrate interactions. We have examined the distribution of PS integrins in wing and leg cell lines, and find that these integrin homologues are expressed preferentially in aggregates. Cell sheets, small cell clumps and chains of cells express antigen at points of cell-cell contact only.
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  • 35
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    Keywords: Drosophila ; Pattern formation ; Segment polarity genes ; gooseberry ; Cell interactions
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    Notes: Summary Segment polarity genes define the cell states that are required for proper organization of each metameric unit of the Drosophila embryo. Among these, the gooseberry locus has been shown to be composed of two closely related genes which are expressed in an overlapping single-segment periodicity. We have used specific antibodies raised against the protein product of the gooseberry proximal (gsb-p) gene to determine the spatial distribution of this antigen in wild type embryos, and to monitor the effects of segment polarity mutants on the pattern of the gsb-p protein distribution. We find that the gsb-p protein accumulates beneath each posterior axonal commissure in the progeny of neuroblasts deriving from the epidermal compartments of wingless (wg) and engrailed (en) expression. The results of this analysis support the idea that gsb-p has a specific role in the control of cell fates during neurogenesis, and indicate that en and wg provide critical positional cues to define the domain in which gsbp will be activated. Furthermore, these data suggest that, in order to be expressed in the embryonic CNS, gsb-p may preliminarily require activity of the gooseberry-distal gene in the epidermis.
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  • 36
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    Development genes and evolution 201 (1992), S. 88-94 
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    Keywords: Fate map ; Drosophila ; Flight muscle ; Mosaics ; Cell lineage
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    Notes: Summary A blastoderm fate map has been prepared for Drosophila, using mosaics of a temperature-sensitive mutation, shibire (shi). The mutation can cause abnormal flight muscle morphology, inducible only by a short heat pulse in early metamorphosis. Thus muscle lineage and development are unperturbed until the heat pulse in the early pupa. The developmental focus of the shi muscle phenotype maps to the ventral thorax at the expected site of thoracic mesoderm, and probably indicates the blastoderm progenitors of the adult flight muscle. The fate map provides greater detail than previously available for the dorsolongitudinal fibers (DLM) of flight muscle, showing wide separation of the fibers of flight muscle. DLM fibers a and b map close together, and far anterior to fibers e and f, which also map together. On a fate map, common developmental focus indicates a common blastoderm origin. Thus, the observed pattern for DLM fibers suggests that the blastoderm progenitors for each of these syncytial fiber pairs (a, b; e, f) include only one or two cells. It follows that there is usually a single genotype within each fiber pair (a, b; e, f), and that this genotype is directly reflected in the fiber phenotype. In a large number of cases, DLM fibers a and b differ in phenotype from other DLM fibers, in parallel with their other differences (e.g., timing of development in pupa, innervation, motor activity). The separation of fate map locations of the developmental focus for DLM fibers within mesoderm suggests that specific fibers of flight muscle may, in normal development, originate in all three thoracic mesodermal parasegments.
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    Development genes and evolution 202 (1992), S. 23-35 
    ISSN: 1432-041X
    Keywords: Ecdysteroid ; Imaginal disc ; Drosophila ; Cell line
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    Topics: Biology
    Notes: Summary We have investigated the action of the moulting hormone 20-hydroxy ecdysone (20-HOE) on our leg and wing imaginal disc cell lines. At the morphological level, cells stop dividing and there is some cell death. The remaining cells elongate and aggregate, often producing long processes which form connections between different aggregates. 20-HOE acts within the first one or two days of a passage, at an optimum concentration of 10 ng/ml, this being about 1/100 of the optimum for ecdysone. One cloned wing cell line, C9, has been found to be relatively insensitive to the action of 20-HOE. We have been able to select for resistance to 20-HOE by growing cells in gradually increasing concentrations of hormone followed by passages in hormone-free medium. This has enabled us to isolate a wing cell line C1.8R from its parent cloned line C1.8+. This shows no response to 20-HOE, and cell growth continues even at hormone concentrations as high as 150 ng/ml. We have measured chitin synthesis by the incorporation of radioactive glucosamine into a cell fraction resistant to extensive alkali hydrolysis. The residue was incubated with chitinase, which resulted in a 50% reduction in labelled product. Treatment with 10 ng/ml of 20-HOE dramatically increased chitin synthesis in line C1.8+, but had no effect in the line C1.8R, selected for resistance to hormone.
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  • 38
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    Development genes and evolution 202 (1992), S. 49-60 
    ISSN: 1432-041X
    Keywords: Drosophila ; Delta ; Enhancers ; Suppressors ; Neurogenesis
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    Notes: Summary We have screened for dominant enhancers and suppressors of the wing phenotype associated with two Delta alleles: Dl 9P39, an amorphic allele, and Dl FE32, an antimorphic allele. The interactions of some of the modifiers with Delta are due to haplo-insufficient expression of the corresponding genes. Although not explicitly shown for the remaining cases, we assume that haploin-sufficiency is also the basis for the relationships of these genes to Delta, since no allele specific interactions were observed. The modifiers found define 22 genes with pleiotropic expression, which can be classified into two groups: genes required for wing vein pattern formation and for neurogenesis, and genes which are not required for neurogenesis. Among the genes of the first group, Hairless and Star were previously known to participate in neural development. One further modifier was found which may correspond to a new neurogenic gene. The second group of genes is larger and includes already known loci, e.g., Plexate, blistered, plexus, etc, as well as other previously unidentified genes, which function during wing morphogenesis.
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  • 39
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    Keywords: Determination ; Germ-line ; Somatic cells ; Inhibitor gradient hypothesis ; Drosophila
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    Notes: Summary A hypothesis is presented which explains the segregation of germ cells from somatic cells, and the subsequent determination of both cell types with a single mechanism. This hypothesis is in part based on that of Meinhardt (1977) and can be summarized as follows: In the newly fertilized egg, the action of a sink in the pole plasm leads to the formation of an anterior-posterior gradient of an inhibitor. The concentration of this inhibitor in the posterior 20% of the egg is below that needed to repress synthesis of an activator. When, during the nuclear division stage, nuclei enter this posterior region, synthesis of the activator begins. As the activator is autocatalytic, this leads to the formation of a peak of activator in this region; and since the activator also catalyses the synthesis of the inhibitor, a peak of inhibitor is formed in the same place. The inhibitor then diffuses anteriorly through the periplasm, forming a posterior-anterior gradient. The presence of this inhibitor in the periplasm causes the nuclei that enter the periplasm to form blastoderm cells and to take up particular segmental states appropriate to their position, while those that remain in the yolk-containing plasm develop into vitellophages. The action of the sink in the pole plasm is postulated to result in the formation of the pole cells, and subsequently to direct some of these into forming cells of the germ-line.
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    Development genes and evolution 189 (1980), S. 1-15 
    ISSN: 1432-041X
    Keywords: Cell line ; Drosophila ; Ecdysone ; Ecdysterone ; Hormones
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    Topics: Biology
    Notes: Summary Cells of the line Kc, derived fromDrosophila melanogaster embryos, extend long processes when exposed to ecdysteroid hormones. We have devised a quantitative assay for this morphological response, using the subline Kc-H. The assay was used to characterize the conditions required for the response. A halfmaximal response is elicited by approximately 10−8M 20-hydroxyecdysone; the response is saturated by 10−7M 20-hydroxyecdysone, which causes detectable elongation within a few hours, and a maximal response after 2–3 days. The response occurs substantially normally in the absence of serum, during growth in suspension, and in over-crowded cultures. It is not elicited by cyclic nucleotides, vertebrate growth factors, or a variety of other non-ecdysteroid reagents. Of 60 ecdysteroid compounds tested, only those which were active in other insect test systems elicited the response, and the concentrations required were approximately proportional to the concentrations active in other in vitro systems. We conclude that the response of Kc cells to 20-hydroxyecdysone retains basic features of the ecdysteroid response of intact tissues and therefore that Kc cells are a useful model system for studying ecdysteroid action.
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  • 41
    ISSN: 1432-041X
    Keywords: Eggshell ; Chorion ; Peroxidase ; Crosslinking ; Drosophila
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    Notes: Summary TheDrosophila chorion contains an endogenous peroxidase activity which remains inactive until late stage 14 when it catalyzes the crosslinking of the chorionic proteins. Using explanted follicles developing in vitro, premature, but otherwise normal crosslinking can be induced with hydrogen peroxide and normal crosslinking can be prevented with peroxidase inhibitors. Inhibition or premature activation of the shell peroxidase allows characterization of chorionic filament specific proteins and establishes new criteria for the identification of eggshell components.
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  • 42
    ISSN: 1432-041X
    Keywords: Early neurogenesis ; Neurogenic mutants ; Drosophila
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    Notes: Summary The central nervous system (CNS) ofDrosophila develops from precursor cells called neuroblasts. Neuroblasts segregate in early embryogenesis from an apparantly undifferentiated ectoderm and move into the embryo, whereas most of the remaining ectodermal cells continue development as epidermal cell precursors. Segregation of neuroblasts occurs within a region called the neurogenic field. We are interested in understanding how the genome ofDrosophila controls the parcelling of the ectoderm into epidermal and neural territories. We describe here mutations belonging to seven complementation groups which effect an abnormal neurogenesis. The phenotypes produced by these mutations are similar. Essential features of these phenotypes are a conspicuous hypertrophy of the CNS accompanied by epidermal defects; the remaining organs and tissues of the mutants are apparently unaffected. The study of mutant phenotype development strongly suggests this phenotype to be due to misrouting into the neural pathway of development of ectodermal cells which in the wildtype would have given rise to epidermal cells, i.e. to an initial enlargement of the neurogenic region at the expense of the epidermogenic region. These observations indicate that the seven genetic loci revealed by the mutations described in this study contribute to control the neurogenic field. The present results suggest that in wildtype development neurogenic genes are supressed within all derivatives of the mesoderm and endoderm and some derivatives of the ectoderm, and conditionally expressed in the remaining ectoderm. The organisation of the neurogenic field in the wildtype is discussed.
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    Development genes and evolution 188 (1980), S. 153-156 
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    Keywords: Drosophila ; Embryogenesis ; mat (3) 1 mutation ; Two-dimensional gels
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    Topics: Biology
    Notes: Summary The synthesis of a protein which has been detected in blastoderm cells but not in pole cells (Gutzeit and Gehring 1979) has been studied further by means of two-dimensional gel electrophoresis. This protein could not be detected at the nuclear multiplication stage. The protein is translated from mRNA which is transcribed at the blastoderm stage since it is not synthesized in detectable amounts when embryos are injected with α-amanitin prior to the blastoderm stage. Also the protein could not be detected when RNA from freshly laid eggs was translated in vitro. Embryos from females which are homozygous for the mutationmat (3) 1 form pole cells but no blastoderm cells (Rice and Garen 1975). Thesemat (3) 1 embryos, as we will call them in this report, express the protein if aged for a period of time sufficient for completion of blastoderm cell formation in control wild-type embryos.mat (3) 1 embryos and embryos injected with α-amanitin show the same syndrome of visible developmental anomalies; however, the studied protein could only be detected inmat (3) 1 embryos but not in α-amanitin injected embryos.
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  • 44
    ISSN: 1432-041X
    Keywords: Drosophila ; Geographic strains ; Chorion proteins ; Electrophoretic variants ; Chorion gene linkage
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    Notes: Summary Drosophila melanogaster chorion proteins are characterized on one-dimensional isoelectric focusing (IF) gels. The six major chorion components previously identified on SDS gels are shown to resolve into at least 11 components in our IF system. IF screening of 102 geographic strains ofDrosophila melanogaster revealed seven cases of variation in major chorion components. Two strains, Crimea and Falsterbo, which were monomorphic for a variant B1 protein and two strains, Skafto and Lausanne, which were monomorphic for a variant C1 protein, were chosen for further study. After IF developmental analysis of F1 hybrids had indicated that the sources of the variation resided in the structural genes for these proteins, each variant was crossed to a multiply marked and inverted strain (BLT) to determine the linkage group of the variant gene. To localize genes to more specific sites multiply marked 3rd (SKERO) or X-chromosomal (CB1) (X-PLE) mapping strains were used. In both Crimea and Falsterbo the gene for the B1 protein is located near map location 26 on the 3rd chromosome. In both Lausanne and Skafto the C1 gene is located on the X chromosome. Hence, for the first time, we have demonstrated genetically the non-linkage of two chorion genes, B1 and C1.
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    Development genes and evolution 189 (1980), S. 147-153 
    ISSN: 1432-041X
    Keywords: Homeotic mutant ; Drosophila ; Clonal analysis ; Timing of gene action ; Determination
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    Topics: Biology
    Notes: Summary Nasobemia (Ns) is a dominant homeotic mutant ofDrosophila melanogaster which converts parts or all of the antenna to mesothoracic leg.Ns has a temperature sensitive period between 48 and 60 h. The hypothesis thatNs acts during this period and is not required thereafter to maintain the homeotic transformation to leg was tested by removingNs fromNs/+ cells at different stages of development through X-ray induced somatic recombination. The expression of theNs homeotic transformation in recombinant wild type (+/+) cells increased sharply between 48 and 65 h. In clones induced after 65 h the expression of the leg transformation was equal in large and small +/+ clones. We interpret these results as supporting the hypothesis that transient action ofNs between 48 and 65 h switches antennal cells to a clonally stable leg determined state whose maintenance does not require futherNs action.
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  • 46
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    Development genes and evolution 201 (1992), S. 1-11 
    ISSN: 1432-041X
    Keywords: Drosophila ; Neurogenesis ; Signals ; Delta ; Notch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ectodermal germ layer of Drosophila melanogaster gives rise to two major cell lineages, the neural and the epidermal. Progenitor cells for each of these lineages arise from groups of cells, whose elements must decide between taking on either fate. Commitment of the progenitor cells to one of the developmental fates implies two factors. One is intrinsic to the ectodermal cells and determines a propensity to take on neural fate; this factor is probably represented by the products of the so-called proneural genes, which are differentially distributed throughout the ectoderm. The other factor in the cells' decision to adopt one of the two alternative fates is intercellular communication, which is mediated by the products of the so-called neurogenic genes. Two types of interactions, one inhibiting and the other stimulating neural development, have been inferred. We discuss here the assumed role of various neurogenic genes, in particular Notch and Delta, in these processes.
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  • 47
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    Development genes and evolution 201 (1992), S. 194-220 
    ISSN: 1432-041X
    Keywords: Drosophila ; Enhancer trap lines ; Embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Embryos of 171 Drosophila lines carrying a P-lacZ insertion on the second or third chromosome were analyzed regarding their pattern of lacZ expression. All lines were selected from a larger screen of about 4000 lines (Bier et al. 1989). Tissue specificity and time of onset of lacZ expression was documented for each line. Thereby, a comprehensive list of markers for the various tissue and cell types of the Drosophila embryo could be assembled. With the help of several P-lacZ lines the development of a number of structures was studied which so far had been described only insufficiently or not at all. In particular, the embryonic origin and early development of the oenocytes, imaginal discs, histoblasts, fat body, dorsal vessel, and perineurial cells was analyzed. Several previously unknown cell types associated with the dorsal vessel, trachea, and epidermis were discovered. By combining data regarding the origin of the different mesodermally derived organs it was possible to generate in some detail a fate map of the mesoderm of the stage 11 Drosophila embryo.
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  • 48
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    Cellular and molecular life sciences 48 (1992), S. 623-629 
    ISSN: 1420-9071
    Keywords: Drosophila ; heat shock ; stress ; heat shock protein ; gene regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Major alterations in genetic activity have been observed in every organism after exposure to abnormally high temperatures. This phenomenon, called the heat shock response, was discovered in the fruit flyDrosophila. Studies with this organism led to the discovery of the heat shock proteins, whose genes were among the first eukaryotic genes to be cloned. Several of the most important aspects of the regulation of the heat shock response and of the functions of the heat shock proteins have been unraveled inDrosophila.
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  • 49
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    Calcified tissue international 32 (1980), S. 229-236 
    ISSN: 1432-0827
    Keywords: Bone ; Ion influxes ; Calcium ; Phosphate ; Exchange
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Quantitative measurements were made of the ion fluxes of calcium and phosphate into and from calvaria (mouse or rat) when clamped in specially designed micro-Ussing chambers. The effects of varying concentrations of calcium were examined on the influx and efflux of calcium and of its counterion, phosphate. A comparable series of experiments was performed with varying phosphate concentrations. Both ions, as their concentrations increased, depressed their own influx, increased their own efflux, and significantly increased the equilibrium concentration, E/K, supported by the calvaria. Similarly, both ions, as their concentrations increased, affected the influx or efflux of their counterion only slightly but did depress the counterion's equilibrium level, E/K, significantly. In spite of these changes it was shown that calvaria effectively buffered the medium at physiological concentrations of calcium and phosphate. The buffering capacity, however, was small, and the balance, E/K, was modified by small uptake or loss of either ion. The small size of the interacting mineral pool was confirmed by direct measurement of the rapidly exchanging fractions of both calcium or phosphate. They were only ∼1% of the total ions present. The significance of these findings is discussed.
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  • 50
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    Calcified tissue international 51 (1992), S. 35-41 
    ISSN: 1432-0827
    Keywords: Kidney ; Endoplasmic reticulum ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A subcellular fraction enriched in cytochrome c reductase (7.9-fold) and relatively de-enriched (0.64-fold) in Na+/K+-ATPase was prepared from canine kidney cortex by sucrose density gradient ultracentrifugation. It was shown by electron microscopy to consist primarily of a light fraction of endoplasmic reticulum (LER). LER vesicles displayed ATP-dependent 45Ca2+ uptake that was insensitive to 10 mM KCN or NaN3, and was promptly released by 20 μM A23187 or ionomycin. Inositol-1,4,5-trisphosphate (IP3) appeared to produce a time-dependent release of 45Ca2+. Vanadate inhibited 45Ca2+ uptake with a Ki≈0.3 mM, further suggesting that the activity resided in the ER rather than the plasma membrane. 45Ca2+ uptake by LER, at 5 μM total [Ca2+], displayed a strong dependence on divalent cations (Mg2+〉Co2+〉Mn2+≫Ba2+≥Cd2+≥Sr2+, present at 2 mM) as well as on monovalent cations (Na+≥K++Na+ 〉K+〉Li+〉choline+), and anions (Cl-〉acetate-≥NO3 -≥F-〉H2PO4 -≫gluconate-≥oxalate=≫SO4 =). It had a fairly narrow pH optimum (7.25–7.50). Preincubation (10 min) of LER vesicles with 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated LER Ca2+ uptake; this effect was enhanced in the presence of renal cytosol [5% (vol/vol)]. However, Ca2+ uptake was not affected by preincubation with dibutyryl cyclic AMP, calmodulin, or 1,25-(OH)2-vitamin D3, either in the absence or presence of renal cytosol. Thus, the Mg2+-ATP dependent 45Ca2+ uptake activity of this canine renal cortical LER fraction displays modulation by IP3, TPA, and pH that appears to be physiologically relevant.
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    Calcified tissue international 51 (1992), S. 143-150 
    ISSN: 1432-0827
    Keywords: Adsorption ; Magnesium ; Calcium ; Apatite crystals ; Enamel ; Dentin ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Magnesium (Mg) is a conspicuous constituent of hard tissues but its possible role in biomineralization is poorly understood. It is possible that Mg2+ adsorbed onto bioapatites may contribute to the modulation of crystal growth as such inhibitory activity has been reported for synthetic apatites. The present study was undertaken to determine the adsorption isotherms of Mg ions onto synthetic apatites and biominerals in tooth and bone tissues in the presence of other ions of natural occurrence. Synthetic crystals used as adsorbents were hydroxyapatite and, as a better prototype for the biomineral, Mg-containing carbonatoapatite. Human enamel and dentin materials were obtained from extracted, caries-free, permanent teeth. Porcine dentin materials at two developmental stages were obtained from erupted deciduous and unerupted permanent teeth of a 6-month-old slaughtered piglet. Porcine bone was obtained from the cortical portion of the mandible of the same animal. All biomineral samples were pulverized and then treated by plasma ashing (deproteination) at about 60°C. Each of the powdered samples was equilibrated in solutions containing various initial concentrations of Mg2+, Ca2+, and Na+ (or K+) as nitrate salts. Following equilibration, concentrations (and activities) of magnesium and calcium ions in the experimental solution were determined. The pH values of the equilibrium solutions were in the range of 6.2–6.5. Experimental data of the Mg adsorption onto hydroxyapatite were interpreted on the basis of a Langmuir-type model for binary systems assuming competition of Mg2+ and Ca2+ for the same adsorption sites on the crystal surfaces of the apatites. According to this model, the adsorbed Mg is expressed as a function of the ionic activity ratio (Mg2+)/(Ca2+) in the equilibrium solution. The model contains two parameters, the adsorption selectivity constant Ks and the maximum number of adsorption sites N (μmol/g). The numerical values of Ks were similar for all adsorbents used (synthetic and biological) and indicated the preferential adsorption of Ca2+ probably due to spacial restrictions extending to the very surface of the crystals. The initial level of Mg2+ in the surface pool was different in the various biominerals, probably reflecting the composition of fluid in which the biominerals were formed. Whereas the surface pool of Mg of human enamel was marginal, only 5% of the total Mg, significant fractions of the total Mg in human and porcine dentins (about 20–30%), and porcine bone (about 40%) existed on the crystal surfaces. There were significant differences in the total Mg and the value of the parameter N between young (unerupted) and mature (erupted) dentin minerals. It was ascertained that the occupancy of adsorption sites by Mg ions became greater with maturation of the dentin tissues. The overall results suggest that the Mg-mineral interaction in tooth and bone tissues may be a highly tissue-specific process, presumably reflecting differences in fluid composition (particularly Ca and Mg activities) responsible for biomineralization.
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  • 52
    ISSN: 1432-0827
    Keywords: Bone cells ; Cyclic AMP ; Calcium ; Ascites fluid resorptive protein
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    Topics: Biology , Medicine , Physics
    Notes: Summary The effects of a bone resorptive protein isolated from human cancer ascites fluid on bone cell calcium and cyclic AMP were studied with fetal rat cells. The osteoclast-activating factor increased bone cell calcium uptake at 37°C and 4°C with no direct effects on calcium efflux. Concentrations of the resorptive factor that increased in vitro bone resorption and cell calcium uptake had no effect on cyclic AMP. The effects of the protein on calcium uptake were not specific for bone cells, and large increases were also observed in isolated fetal rat skin cells. These studies suggest that increases in the permeability of the cell membrane to calcium are involved in the mechanism of action of the ascites fluid resorptive protein.
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  • 53
    ISSN: 1432-0827
    Keywords: Hydroxyapatite ; Dissolution ; Pyrophosphate, Diphosphonates ; Calcium ; Phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Pyrophosphate and diphosphonate ions have been said to diminish the dissolution of hydroxyapatite crystals, because they lower the equilibrium concentrations of calcium and phosphate ions in the bulk solution around hydroxyapatite crystals in a closed system. However, in a closed system these effects are not necessarily due to an effect on dissolution alone. In this paper we have used a continuous flow system to study the effects of pyrophosphate and two diphosphonates, ethane-1-hydroxy-1,1-diphosphonate and dichloromethane diphosphonate, on the dissolution of hydroxyapatite. All three compounds decreased markedly the rate of dissolution of hydroxyapatite as well as the exchangeable pools of calcium and phosphate ions around the cystals.
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  • 54
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    Calcified tissue international 31 (1980), S. 231-238 
    ISSN: 1432-0827
    Keywords: Magnesium ; Bone ; Calcium ; Parathyroid gland
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    Topics: Biology , Medicine , Physics
    Notes: Summary Calcium homeostasis and bone pathology were studied in weanling rats fed a low (70 ppm) magnesium diet for 2–21 days. The rats developed significant, progressive hypercalcemia after 6 days on the diet. The increase in blood calcium was accompanied by progressive hypoactivity of the parathyroid gland (PTG), as determined by histologic and morphometric analyses. Thus hyperactivity of the PTG could not have been responsible for the hypercalcemia observed. Histologic examination of femora and humeri from magnesium-deficient rats showed progressive subperiosteal hyperplasia, consisting of undifferentiated osteoprogenitor cells and fibrous tissue, after 7 days of deficiency. The presence of unmineralized osteoid tissue in the metaphyses indicated that mineralization was not proceeding normally. The alterations in differentiation of osteoprogenitor cells, together with the failure of mineralization, resulted in significantly lower rates of bone formation (as measured by fluorochrome labeling) in the magnesium-deficient rats. Basophilic cementing lines and inactive osteocytes in the cortices of bones from magnesium-deficient rats indicated that bone resorption was also severely reduced in magnesium deficiency. We postulate that bone magnesium depletion (66% by day 21) has a direct negative effect on osteoblastic and osteocytic activity, and may explain, in part, the decreased responsiveness of bone to parathyroid hormone (PTH) that has been observed in magnesium-deficient animals.
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    Calcified tissue international 32 (1980), S. 45-53 
    ISSN: 1432-0827
    Keywords: Dentin ; Periodicity ; Allometry ; Calcium ; Sulfur
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    Topics: Biology , Medicine , Physics
    Notes: Summary We have described differences in the aspects of biological rhythms for calcium and sulfur deposition on the labial and lingual sides of the growing rabbit incisor, where growth occurs along a spiral axis. The calcium oscillations appear to be smoother on the labial side than on the lingual side. The lingual side is characterized by high-frequency rhythms with high amplitudes which possess the greatest percent of the power (Fourier analysis). These observations also reflect a difference in behavior of the mean Ca concentration across the labial and lingual sides. Sulfur rhythms on the labial side have higher amplitudes than those on the lingual side, but systematic differences in distribution of power between high and low frequencies is not as pronounced as in the case of Ca. The differences in Ca rhythms reflect differences in the growth rates of incisors on either side of the spiral axis. The labial side grows slightly faster than the lingual side, and its odontoblasts secrete Ca along the spiral axis and toward the pulp cavity at the same time. Thus the resultant direction of growth is more nearly opposite the extension of the occlusal end on the labial side, and Ca is consequently deposited over a wider area relative to that on the lingual surfaces. On the lingual side, Ca is deposited within a more limited area, and growth must therefore be continuous at high frequencies. The distribution of Ca on both sides of the tooth reflects these differences in growth rate and periodicity in two ways. First, given a unit area of tooth, the calcium concentration on the labial side is less than that of the lingual side. Second, whereas the calcium concentration on the labial side declines rapidly from the enamel-dentin junction to the pulp cavity, it is uniformly high across the lingual side because its growth is more continuous at high frequencies.
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  • 56
    ISSN: 1432-0827
    Keywords: 1α-Hydroxy vitamin D3 ; 1,25-Dihydroxyvitamin D3 ; Calcium ; Transport ; Intestine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The timed sequence of events following the oral administration of 1α-hydroxy vitamin D3 (1αOHD3) to rachitic chicks was compared to that following a comparable dose of 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3). RNA polymerase activity was maximally increased 20% by 1αOHD3 within 1 to 2 h and returned to control values after 8 h. Alkaline phosphatase activity was stimulated by 4 h and was maximal (3- to 5-fold increase) at 24 h. Calcium binding protein (CaBP) was detected initially within epithelial cells at the proximal end of the villus (just above the crypt) 6 to 8 h after 1αOHD3 administration, in epithelial cells lining the proximal half of the villus by 24 h, and in epithelial cells along nearly the entire villus by 48 h. At no time did goblet cells contain CaBP. Serum calcium concentrations were significantly elevated in 2 h and maximal by 12 h (an increase of 3.6 mg/dl). Calcium accumulation by the intestinal mucosa in vitro was increased by 6 to 8 h and maximal (60% increase over controls) at 24 h. Phosphate accumulation by the intestinal mucosa in vitro was increased by 6 h and maximal (105% increase over controls) between 8 and 24 h. 1,25(OH)2D3 increased CaBP and calcium accumulation by 4 h, 2 h sooner than did 1αOHD3. 1,25(OH)2D3 decreased serum calcium levels and increased serum phosphate levels at 2 h unlike 1αOHD3. No difference in the effects of these compounds on alkaline phosphatase activity, RNA polymerase activity, and phosphate accumulation could be demonstrated. These results are consistent with the possibility that 1αOHD3 may not require conversion to 1,25(OH)2D3 for all of its biological effects.
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  • 57
    ISSN: 1432-0827
    Keywords: Perifusion ; Parathyroid hormone ; In vitro ; Calcium ; Beta regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary An in vitro perifusion system was used to study parathyroid hormone (PTH) secretion in response to calcium (Ca) and beta-adrenergic agents. Perifused parathyroid tissue responded to changes in Ca within the physiologic range during experiments up to 5 h. There was rapid secretory stimulation after exposure to low Ca, with the maximum response being observed at 20 min. Normal bovine glands showed a Ca-independent nonsuppressible component of PTH release at concentrations of Ca above physiologic. 1-isoproterenol produced rapid stimulation of PTH release, the response being blocked by a beta antagonist. The maximum secretory response to either low Ca (0.5 mM) or 1-isoproterenol (10−5 M) was enhanced when the two stimuli were applied simultaneously. The response to isoproterenol was blocked by raising Ca to 2.5 mM. Although d,l-propranolol (10−4 M) caused mild suppression of PTH release at a Ca of 1.25 mM, it did not cause additional suppression at 2.5 mM Ca nor did it decrease the response to 0.5 mM Ca stimulation. The secretory response of the gland to low Ca was sustained at a level more than double the baseline rate. The response to isoproterenol was more transient, with a return to or toward baseline secretion within 60 min. These results suggest that beta agonists and low Ca have separate but related mechanisms for stimulating PTH release and may affect different pools of hormone. The perifusion system described is a relatively simple technique for assessing the kinetics and interactions of various stimulators of PTH secretion.
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    Calcified tissue international 32 (1980), S. 29-44 
    ISSN: 1432-0827
    Keywords: Rabbit ; Dentin ; Calcium ; Sulfur ; Periodicity ; Circadian ; Ultradian
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We have identified a variety of biological rhythms involved in the apposition and mineralization of dentin in the rabbit incisor. Animals were injected during the day or night with lead acetate at 2-week intervals—to provide biological time markers in forming dentin—and transverse undecalcified sections of the lower incisors were prepared for electron microprobe analysis. The positions of the lead markers were identified, and the continuous distribution of calcium and sulfur was measured at 1 µm intervals between the markers. In thin sections stained with hematoxylin after decalcification, the widths of a series of structural increments (bands) were measured with an ocular micrometer. Fourier analysis of the data revealed spectra of structural and compositional rhythms with a range of periodicities which extended from a matter of hours [ultradian (〈24 h)] to days [infradian (〉24 h) and circadian (approximately 24 h)]. The structural and compositional rhythms appeared to be independent to the extent that they did not necessarily have the same periods, or amplitudes. Nor were there simple phase relationships between all of the rhythms. At some times, Ca and S fluctuations are inversely proportional (180° out of phase), but in other cases they are directly proportional or out of phase by varying degrees other than 180°. The analyses thus suggest that calcium and sulfur deposition (representing mineral and glycosaminoglycan deposition, respectively) are not simply inversely proportional, and that the hematoxylin-stained structural increments did not solely reflect differences in the distribution of the mineral components in dentin.
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  • 59
    ISSN: 1432-0827
    Keywords: Menopause ; Estrogens ; Bone ; Osteoporosis ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Biochemical parameters reflecting bone resorption [urinary calcium/creatinine (Ca/Cr) and hydroxyproline/ creatinine (OH/Cr)] were related to serum estrogens [estrone (E1) and estradiol (E2)] in 262 healthy women including 158 patients receiving estrogen replacement therapy (ERT) for at least 6 months, 49 eugonadal women, and 55 untreated postmenopausal women. A significant (P〈0.001) correlation exists between serum E2 and Ca/Cr: Ca/Cr (mg/dl)=-0.00044 E2 (pg/ml)+0.129 (n=262; r=-0.37), serum E2 and OH/Cr: (OH/Cr (mg/g)=-0.049 E2 (pg/ml)+18.76 (n=262; r=-0.36), serum E1 and Ca/Cr: Ca/Cr (mg/dl)=-0.0003 E1 (pg/ml)+0.127 (n=261; r=-0.28) but not between serum E1 and OH/Cr. Women with circulating levels of E2 between 60 and 90 pg/ml have a significant (P〈 0.01) reduction of Ca/Cr and OH/Cr when compared with those with lower levels of E2. Higher values of E2 do not provide additional benefit. We conclude that in postmenopausal women receiving an estrogen replacement therapy (ERT), a significant reduction of bone resorption is achieved when circulating levels of estradiol reach a value (60 pg/ml) corresponding to the one measured, in eugonadal women, during the last days of the early follicular phase of the menstrual cycle. We suggest that oral or percutaneous ERT should induce a minimal value of 60 pg/ml to prevent postmenopausal bone loss.
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    Calcified tissue international 30 (1980), S. 27-34 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Calcium ; Cartilage ; Vesicles
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    Topics: Biology , Medicine , Physics
    Notes: Summary The potassium pyroantimonate technique was utilized for the selective subcellular localization of calcium in the mandibular condylar cartilage of 1-day-old rats. Electron dense calcium pyroantimonate precipitates were localized principally in mitochondria and at the cell membrane of the chondrocytes. In addition, small intracellular vesicles 0.1–0.2µm in diameter were observed in proximity to the cell membrane of chondrocytes of the mid-hypertrophic zone. The results suggest that these vesicles were being extruded from the cell into the extracellular matrix. Energy-dispersive analysis by X-rays confirmed that calcium is the principal cation of the electron-dense precipitates.
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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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  • 62
    ISSN: 1432-0827
    Keywords: 31P NMR spectroscopy ; Phosphoprotein ; Dentin ; Calcium ; Inorganic orthophosphate ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The single phosphoprotein of fetal calf dentin, having a molecular weight of approximately 94,000 and a phosphorus content of 8% (w/w), was examined by31P NMR spectroscopy. The single resonance at 3.7 ppm at pH 10 and its chemical shift during acid titration established the phosphomonoester nature of the organic phosphorus moiety. During titration of the phosphoprotein with CaCl2 in the presence of inorganic orthophosphate ions, line broadening for the orthophosphate resonance was both phosphoprotein- and calcium-dependent, indicating ternary complex formation. The data indicate that the phosphoprotein of fetal calf dentin binds both calcium and inorganic orthophosphate ions and therefore has the requisite physical chemical properties necessary for it to facilitate the heterogeneous nucleation of a Ca-PO4 solid phase from solution during tissue mineralization.
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  • 63
    ISSN: 1432-2048
    Keywords: Calcium ; Chloride ; Epidermis (leaves) ; Ion compartmentation ; Mesophyll
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cation and anion distribution between the epidermis and mesophyll of primary leaves of 10-d-old barley (Hordeum vulgare L.) seedlings was studied in relation to growth conditions. A new method was employed to isolate epidermal protoplasts. The following observations were made: (i) Under standard hydroponic growth conditions, K+ was the dominant cation and NO 3 3− the predominant anion, both in epidermal and mesophyll protoplasts. (ii) Levels of Cl− and particularly of Ca2+ were specifically increased in the epidermis when growth conditions stimulated uptake of these ions into the plants. (iii) Epidermal PO 4 3− was maintained at a very low level even in the presence of 50 mM phosphate in the rooting medium, whereas the phosphate concentration in the mesophyll was high, with phosphate being accumulated in the mesophyll vacuoles. (iv) Sulphate and NO 3 − accumulated to a similar degree in the mesophyll and the epidermis when K2SO4 or KNO3 in the rooting medium caused salt stress to the plants. (v) Epidermal protoplasts took up 35SO 4 2− and 36Cl− from the surrounding medium. A comparison of the uptake rates indicates that specific ion deposition into the epidermis may partially be the consequence of differential uptake of ions from the transpiration stream. The results indicate an important function of the epidermis for ion compartmentation in barley leaves.
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    Planta 188 (1992), S. 279-288 
    ISSN: 1432-2048
    Keywords: Aplanospore (germination) ; Calcium ; Vaucheria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Germination of aplanospores in Vaucheria longicaulis Hoppaugh var. macounii Blum proceeds through three stages of development. Stage I begins with the initiation of germination and lasts approx. 2 h. During this stage germinating filaments grow at an accelerated rate (266 ± 12 μm · h−1). Stage II is characterized by a sharp decline in the growth rate of germinating filaments (96 ± 4 μm · h−1) and lasts 4 h. This is followed, during the next 4 h, by a recovery in the growth rate (168 ± 8 μm · h−1) of germinating filaments, stage III. Growth rates stabilize and remain unchanged during subsequent development (Oliveira and Fitch, 1988, J. Submicrosc. Cytol. Pathol. 20, 397–406). The Ca2+-influx modulators LaCl3, nifedipine and methyl 1,4-dihydro-2,6-dimethyl-3-nitro-4 (2-trifluoromethylphenyl)-pyridine-5-carboxylate (Bay K-8644), the ionophore calcimycin (A23187), the intracellular Ca2+-release antagonist 8-N-N'-(diethylamino)-octyl-3,4, 5-trimethoxybenzoate (TMB-8), the Ca2+-uptake inhibitor ruthenium red and the phosphoinositide-cycle modulators LiCl and myo-inositol show that the events required for the initiation are distinct from those required for the completion of each stage of germination. These studies in conjunction with microinjection of germinating filaments with inositol 1,4,5-trisphosphate, the natural ligand for Ca2+ release from Ca-storing organelles (endoplasmic reticulum, vacuole), and treatment with chlorotetracycline (CTC), to visualize the distribution of membrane-bound Ca2+ reveal that both the initiation and completion of each stage of germination are controlled by Ca2+ signals which are restricted to well-defined time intervals and are modulated by the origin (source) of Ca2+.
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    Planta 158 (1983), S. 320-328 
    ISSN: 1432-2048
    Keywords: Calcium ; Egeria ; Fluorescent probe ; Ions, group II ; Symplast
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    Topics: Biology
    Notes: Abstract The hydrophyllic dyes fluorescein glutamic acid, fluorescein glutamylglutamic acid (F(Glu)2), fluorescein hexaglycine, fluorescein leucyldiglutamyl-leucine and 6-carboxyfluorescein are unable to pass the plasmalemma in leaves of E. densa. However, when injected into single cells the dye conjugates of molecular weight 665 dalton or less move freely from cell-to-cell. This intercellular movement presumably occurs via the plant symplast. Movement of F(Glu)2 from the injected cell occurs with greatly reduced frequency when Ca2+, Mg2+ or Sr2+ are injected into the cell immediately prior to the dye. The fraction of dye injections leading to movement declines with increasing group II ion concentration in the electrode tip, up to 10 mM. Sodium and K ions do not affect dye movement. When dye injection is delayed 30 min after Ca2+ injection, dye movement is no longer inhibited. Thus the cells recover from the Ca2+ injection, indicating that the ion does not cause major cell damage. Recovery from Mg2+ injection is not complete within 60 min. Treatment of leaves with chemicals expected to raise the concentration of free intracellular group II ions, notably the mitochondrial uncoupler carbonyl cyanide p-trifluoromethoxyphenyl hydrazone, the inhibitor of mitochondrial Ca2+ uptake trifluralin, or the ionophore A23187 also inhibits dye movement, while the calmodulin inhibitor trifluoperazine does not. Cytoplasmic streaming is inhibited by Ca2+ or Mg2+ injection and by the metabolic inhibitors. However when streaming is stopped by cytochalasin B, dye movement is not inhibited. Hence steaming is not necessary for dye movement. Thus the cytoplasmic concentration of free group II ions may directly regulate the permeability of the plant symplast.
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  • 66
    ISSN: 1432-1939
    Keywords: Wood mouse ; Bank vole ; Calcium ; Growth ; Body weight
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    Topics: Biology
    Notes: Summary Calcium intake by wild rodents varies with season and habitat. This may have important ecological consequences; several studies have suggested that calcium availability may limit growth and reproduction. We studied the effect on growth of varying the calcium intake of captive wood mice Apodemus sylvaticus (L.) and bank voles Clethrionomys glareolus (Schreber 1780). To determine whether effects observed in the laboratory could be detected in free-living animals, we also compared the body weights and lengths of free-living animals on calcium-poor gritstone areas with those of rodents on calcium-rich limestone habitats where the dietary calcium concentration was between 2 and 5 times higher. Captive wood mice fed high calcium (0.87%) diet grew at the same rate as mice fed low calcium (0.30%) diet but continued growing for longer, thereby achieving higher asymptotic weights. In contrast, captive bank voles fed the high calcium diet grew more slowly and had lower asymptotic weights than voles fed the low calcium diet. As expected from the laboratory growth study, the higher calcium intake of free-living wood mice on the limestone was associated with greater body size compared with mice on the gritstone. However, bank voles were also larger on the limestone, even though high calcium intake impaired growth in captive animals. The contrast between wood mice and bank voles in the effects of calcium on growth, the reason why impaired growth may not be apparent in bank voles from calcium-rich habitats and the ecological significance of these results are discussed.
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  • 67
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    Oecologia 92 (1992), S. 183-187 
    ISSN: 1432-1939
    Keywords: Drosophila ; Resource exploitation ; Decaying-herbage breeding ; Host choice
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    Topics: Biology
    Notes: Summary The Drosophila fauna of a deciduous flood plain forest rich in undergrowth near the river Isar, close to Munich, Germany, was surveyed in summer 1990. Decaying herbage baits (decay artificially induced) were set out to study the exploitation of that resource by Drosophila. Sixteen plant species belonging to several families dominant in the collecting area were tested. All attracted and produced drosophilid flies. Ten Drosophila species utilized decaying plant material as breeding sites; at least eight of the ten are polyphagous. Decaying stalks and leaves of Angelica sylvestris (Apiaceae) were examined in detail. In the case of the most frequent species of Drosophila attracted to A. sylvestris, the number of adults collected did not correlate with the number of flies emerging from the substrate. This was particularly true of D. limbata and D. phalerata. When oviposition and larval development of D. limbata and D. phalerata on A. sylvestris was tested in the laboratory, the number of offspring per female was the same in both species. The difference between these two species of the quinaria group in the exploitation of A. sylvestris in the field is therefore not due to differential suitability of the substrate.
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  • 68
    ISSN: 1432-2145
    Keywords: Calcium ; Calmodulin ; Microsporogenesis ; Pollen germination ; Gasteria
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    Topics: Biology
    Notes: Summary The distribution of membrane calcium and calmodulin (CaM) has been fluorimetrically determined in the anther of Gasteria verrucosa with particular attention to sporogenous cells, meiocytes, microspores, pollen and stages of pollen germination and tube growth using chlortetracycline (CTC) and fluphenazine (FPZ). CTC and FPZ fluorescence in sporogenous cells is relatively higher than in the adjacent tapetal cells, indicating higher membrane calcium and CaM levels in the former cell type. However, during meiosis there is a significant increase in membrane calcium and CaM levels in the meiocytes compared to that found in the young microspores. CTC and FPZ fluorescence in the sporogenous cells, meiocytes and young microspores is punctate and slightly diffused throughout the cytoplasm. In the microspores of the tetrad and the young released microspores CTC fluorescence (CTCf) is polarized and mainly associated with the area opposite the future colporal region. FPZ fluorescence (FPZf) becomes polarized in the young microspore. Subsequently, there is a shift in the polarity, and most of the CTCf and FPZf in the old microspores and pollen is regionalized towards the colporal region, and the fluorescence is more diffused, indicating a change in the organellar-bound calcium and CaM. This final graded distribution of CTCf is maintained during pollen germination in that the growing pollen tubes invariably show a tip to base membrane-calcium gradient. In the tapetal cells a high level of Ca2+ is present during the microspore stage. During the preparation for anthesis the endothecium differentiation is marked by the presence of Ca2+. Post-treatment of labelled cells with a Ca2+ chelator such as EGTA resulted in a substantial decrease in diffuse and punctate CTCf. Alternatively, treatment of cells with non-ionic detergent Nonidet P-40 resulted in the total elimination of CTCf, suggesting that the observed CTC fluorescence was due to membrane-associated calcium. The cytological specification of CTC as a probe for calcium is discussed. From cytofluorometric measurements and atomic absorption, it became clear that the level of Ca2+ in the anther is high during the sporogenous and meiotic phases. An increase in CTCf and FPZf occurred after microspore mitosis. An interaction of Ca2+ transport from tapetum to the young pollen is postulated. These findings suggest that the level of Ca2+ in the anther during meiosis is generally relatively higher than at the sporogenous or young microspore stage. These findings are discussed in the light of available information on the role of Ca2+ and CaM-mediated processes such as cell division, callose synthesis and pollen-tube tip growth.
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  • 69
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    Sexual plant reproduction 5 (1992), S. 138-145 
    ISSN: 1432-2145
    Keywords: Pollen germination ; ATPases ; Calcium ; Magnesium ; Potassium ; Agapanthus umbelatus
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    Topics: Biology
    Notes: Summary Cytochemical detection of ATPase activity in the pollen grain (PG) and pollen tube (PT) of Agapanthus umbelatus showed that the enzymes concerned presented specific patterns of membrane distribution according to their ionic dependencies and to the timecourse of germination and tube growth. In the pollen tubes Ca2+-ATPases were mainly localized in mitochondria and ER membranes, while Mg2+-ATPases were found especially in the tonoplast and in the membrane of the P-particles. K+-ATPases showed a high activity at the plasma membrane. In the pollen grain similar patterns of ATPase activity were observed. The highest activity of all three types was observed at the plasma membrane of the grain and at the intine and inner exine layers of the cell wall. The activity observed in the pollen grain cell wall decreased with germination time. In vivo germination studies in the presence of specific inhibitors of the ATPases showed patterns of inhibition that could be correlated with the corresponding ATPase putative role. The results are discussed in terms of the ultrastructural organization of the PG and PT, especially those correlated with (1) formation and maintenance of ionic gradients throughout the PT, (2) polarized growth and (3) hydrodynamics of PT elongation.
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  • 70
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    Sexual plant reproduction 5 (1992), S. 151-155 
    ISSN: 1432-2145
    Keywords: Calcium ; Chlorotetracycline ; Fertilization ; Nicotiana ; Synergid degeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Synergid degeneration was examined in the isolated embryo sac and egg apparatus of Nicotiana tabacum using quantitative cytology, fluorochromatic reaction (FCR) and chlorotetracycline (CTC). Most synergid degeneration occurs after pollen tubes (PT) arrive in the ovarian chamber between 42 and 48 h after pollination; synergid degeneration was precluded when PT were prevented from entering the ovary by stylar excision indicating that the signal that triggers synergid degeneration travels only relatively short distances in this plant. There was no evidence for any preferentiality between right or left synergids with regard to cell size or degeneration pattern. FCR staining confirms that synergid degeneration involves the loss of membrane integrity and is a reliable indicator of the onset of degeneration. CTC labeling of the degenerated synergid reveals that a concentrated reserve of membrane-bound calcium is present in the receptive synergid, possibly aiding in the attraction, arrest and discharge of the PT, releasing the sperms into the receptive ES.
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  • 71
    ISSN: 1432-072X
    Keywords: Phototaxis ; Motility ; Flagellate ; Chlamydomonas ; Sodium azide ; Calcium ; A 23 187
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    Topics: Biology
    Notes: Abstract A fully automated and computerized method for simultaneous measurements of motility and phototaxis of unicellular flagellates is described. Both systems are directly coupled with a homocontinuous culture. The motility measuring apparatus is equipped with a video camera and recorder for simultaneous single cell behaviour studies. First results of studies on the effects of the phototaxis inhibitor sodium azide and the Ca2+ conducting ionophore A23 187 on motility and phototaxis of Chlamydomonas are reported and correlated with video observations. These results demonstrate that the described systems give informations of whether phototaxis or motility or both are inhibited by chemicals.
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    Archives of microbiology 158 (1992), S. 85-92 
    ISSN: 1432-072X
    Keywords: Arthrobacter photogonimos ; lipA Gene ; Photoinduction ; Calcium ; Metals ; Chelation ; Gene expression
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    Topics: Biology
    Notes: Abstract Expression of the light-inducible (lipA) gene in Arthrobacter photogonimos is repressed by Ca2+ at a concentration greater than 0.1 μM. Expression of lipA was induced by relatively high concentrations of Zn2+ Ni2+ or Co2+ in cell suspensions, an effect that was blocked by an increase in the concentration of Ca2+ in the medium. Zn2+ and other metals apparently overcame repression by Ca2+ by competing for a cellular binding site. Expression of lipA was also induced when the amount of free Ca2+ was lowered with ethylene-bis (oxyethylenenitrilo)tetraacetic acid (EGTA). Our results show that the lipA gene does not require Zn2+ or other divalent cation for expression and that it is regulated negatively by Ca2+. Accumulation of the mature product of this gene (light-inducible protein, LIP) was minimal in the presence of EGTA. Accumulation increased 10-to 20-fold when divalent cations such as Ca2+, Mn2+, Cu2+ or Zn2+ were added to cell suspensions treated with chelator. These divalent cations, which allowed the protein to achieve a protease-resistant form on the cell surface, could be substituted by protease inhibitors such as antipain, leupeptin or 1,10-phenanthroline. Our data can be explained by a biparous mechanism in which divalent cations regulate both expression of the lipA gene and accumulation of the gene product.
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  • 73
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    Biochemical genetics 21 (1983), S. 199-211 
    ISSN: 1573-4927
    Keywords: Drosophila ; lactate dehydrogenase ; isozymic pattern ; development ; isozymic conversion
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Partially purified lactate dehydrogenase (LDH) from third-instar larvae displays two bands (one major and one minor) on polyacrylamide gels. Analogous preparations from pupae and adults exhibit three LDH-staining bands (one major and two minor) in a similar pattern. The migration of the major band is similar for larvae, pupae, and adults, while the two minor LDH bands of pupae and adults migrate more slowly than the minor larval band. It has been shown that larval LDH incubated with β-nicotinamide adenine dinucleotide exhibits two additional minor bands with an electrophoretic mobility similar to that of the minor bands of both pupae and adults. The intensity of the minor larval LDH band (exhibited also by untreated preparations) is drastically reduced. This fact indicates that the life-cycle stage-dependent LDH isozymic distribution is possibly due to a posttranslational effect(s). Highly purified LDH from larvae, pupae, or adults, obtained by an affinity chromatography procedure, displays just one dispersed band, located in the area between the band 5 and the band 6 exhibited by crude extract preparations. These data, in combination with the lack of difference in catalytic properties among enzymes from larvae, pupae, and adults, suggest that LDH synthesis is controlled by the same single structural gene at all developmental stages.
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    Biochemical genetics 21 (1983), S. 199-211 
    ISSN: 1573-4927
    Keywords: Drosophila ; lactate dehydrogenase ; isozymic pattern ; development ; isozymic conversion
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Partially purified lactate dehydrogenase (LDH) from third-instar larvae displays two bands (one major and one minor) on polyacrylamide gels. Analogous preparations from pupae and adults exhibit three LDH-staining bands (one major and two minor) in a similar pattern. The migration of the major band is similar for larvae, pupae, and adults, while the two minor LDH bands of pupae and adults migrate more slowly than the minor larval band. It has been shown that larval LDH incubated with β-nicotinamide adenine dinucleotide exhibits two additional minor bands with an electrophoretic mobility similar to that of the minor bands of both pupae and adults. The intensity of the minor larval LDH band (exhibited also by untreated preparations) is drastically reduced. This fact indicates that the life-cycle stage-dependent LDH isozymic distribution is possibly due to a posttranslational effect(s). Highly purified LDH from larvae, pupae, or adults, obtained by an affinity chromatography procedure, displays just one dispersed band, located in the area between the band 5 and the band 6 exhibited by crude extract preparations. These data, in combination with the lack of difference in catalytic properties among enzymes from larvae, pupae, and adults, suggest that LDH synthesis is controlled by the same single structural gene at all developmental stages.
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  • 75
    ISSN: 1573-4927
    Keywords: Drosophila ; alcohol dehydrogenase ; enzyme polymorphisms, activity ratios
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Representatives of five allozymic classes of Drosophila alcohol dehydrogenase have been compared with respect to their activity levels on two alcohol substrates, quantities of ADH protein, and stability in crude extracts. Within each allozymic class, strains from widely diverse geographic locations differ in their enzyme activity levels but are identical for a measure known as “activity ratio,” which is obtained by dividing the average activity reading on isopropanol by that obtained with ethanol. They are also similar in the rate at which ADH activity declines in crude extracts held at 25°C. For several of the fast-resistant and fast-moderate strains, differences in ADH activity are associated with differences in the amount of enzyme present. The catalytic efficiencies of the fast-resistant forms are considerably lower than those of the fast-moderate allozymes. The origin and persistence of the rare but ubiquitous fast-resistant allozyme is discussed.
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  • 76
    ISSN: 1573-4927
    Keywords: Drosophila ; ontogeny ; amylase ; α-glucosidases ; functional significance
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Changes in amylase (E.C. 3.2.1.1), maltase (E.C. 3.2.1.20), sucrase, and PNPGase activities in relation to changes in wet weight and protein content were studied during the development of larvae and adult flies from two strains of Drosophila melanogaster, homozygous for different amylase alleles. All α-glucosidase activities increase exponentially during a large part of larval development, parallel to the increase in weight, and drop at the end of the third instar. Amylase activity of the Amy 1 strain follows the same pattern. In contrast, amylase activity of the Amy 4,6 strain continues its exponential increase longer. In the third larval instar amylase activity in the Amy 4,6 strain becomes much higher than in the Amy 1 strain. During the first hours of adult life amylase activity of the two strains does not differ. Then Amy 4,6 activity starts to rise and becomes much higher (4–5 times) than Amy 1 amylase activity, which remains approximately constant. All adult enzyme activities are much higher than in larvae. Comparison of enzyme activity of amylase and α-glucosidases in larvae and adults confirms that differences in amylase activities can become important only when starch is a limiting factor in the food.
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    Biochemical genetics 18 (1980), S. 439-454 
    ISSN: 1573-4927
    Keywords: Drosophila ; acetylcholinesterase ; insecticide resistance ; electrophoretic variation
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We examined the Canton-S strain of Drosophila melanogaster for electrophoretic variation of the enzyme acetylcholinesterase. The pattern of bands obtained (stained with acetylthiocholine) depended on age, sex, and tissue (i.e., head vs. body part and hemolymph). However, through mixing experiments, it was concluded that most of these apparent differences were due to modification of the enzyme by unknown substances located in the fly's body. The electrophoretic pattern of head acetylcholinesterase was altered so that it became characteristic of the body which was present during extraction. For example, when heads of D. melanogaster were homogenized in an extract from D. lebanonensis bodies, the characteristic AChse bands of melanogaster were absent and instead the bands of lebanonensis were found. it was found that extraction of adult heads in 0.1 M potassium phosphate buffer alone or with a 2-min exposure to 1 mg/ml trypsin at 20 C gave the most reproducible results, independent of age and sex. Using these conditions, 25 strains of D. melanogaster and 30 strains of D. pseudoobscura were examined without finding any reproducible electrophoretic variant of acetylcholinesterase. In addition, 53 strains from 39 other Drosophila species produced a total of only six electrophoretic forms of the head enzyme. Additional electromorphs were found when whole flies were used, but these were not studied in detail because of the possibility that they could be due to postextraction modification.
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    Biochemical genetics 18 (1980), S. 717-726 
    ISSN: 1573-4927
    Keywords: Drosophila ; 5-fluorouracil ; drug response ; thymidylate synthetase
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Mutant strains sensitive and resistant to the drug 5-fluorouracil (FU) have been isolated from the wild-type Pac strain of Drosophila melanogaster. The resistant strain, termed flur, is resistant to at least 0.0035%FU (2.7 × 10−4 m) in the food media and exhibits cross-resistance to 5-fluorodeoxyuridine (FUdR) but not to 5-fluorouridine (FUR). The sensitive strain termed flu S , exhibits over 90% mortality on 0.0008% FU (6 × 10−5 m). Genetic analysis indicates that the flu gene is located on the third chromosome, which agrees with results of previous workers. An analysis of the enzyme thymidylate synthetase from the selected sensitive and resistant strains indicates that the resistant strain enzyme possesses an elevated specific activity. Levels 4 times that of the sensitive strain were observed when the enzymes were assayed at 20 C. This increase is apparently not due to induction by FU in the food media. It is suggested that the enzyme thymidylate synthetase may be involved in the resistance process.
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  • 79
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    Biochemical genetics 18 (1980), S. 781-791 
    ISSN: 1573-4927
    Keywords: Drosophila ; phenylalanine hydroxylase ; developmental regulation
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Herein we demonstrate that Drosophila larvae possess a synthetic activity capable of converting phenylalanine to tyrosine. This system is readily extractable and displays many characteristics of phenylalanine hydroxylase systems described in other organisms, the most notable being that a tetrahydropteridine is required for full expression of activity. The level of phenylalanine hydroxylase activity present in the organism varies with the stage of development: from an undetected level of activity at the first larval instar, there is a rapid increase in phenylalanine hydroxylase activity which reaches a peak at the time of puparium formation, after which there is a rapid decrease again to an undetected level.
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  • 80
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    Biochemical genetics 18 (1980), S. 929-937 
    ISSN: 1573-4927
    Keywords: glucose oxidase ; glucose metabolism ; Drosophila ; sex specific
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A glucose oxidase (GO) has been identified in the ejaculatory duct of male Drosophila melanogaster. Evidence is given that this enzyme was previously misidentified as HEX-1. Genetic analysis indicates that the Go structural gene is located on the third chromosome at 48 ± 0.5 cm. Go is polymorphic in males in populations of D. melanogaster and D. simulans located in Athens, Georgia. Two other hexose enzymes have also been tentatively identified for the first time in Drosophila. These are NAD(P)-glucose dehydrogenase (GODH) and NAD-gluconate dehydrogenase (GNDH). GODH and GNDH are found in both males and females and may circumvent the initial steps in the pentose shunt.
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    Biochemical genetics 21 (1983), S. 703-711 
    ISSN: 1573-4927
    Keywords: Drosophila ; glucose-6-phosphate dehydrogenase ; polymorphism ; sequential electrophoresis
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Studies were undertaken to investigate two critical aspects of the glucose-6-phosphate dehydrogenase polymorphism in Drosophila melanogaster. The first investigation unequivocally maps the genetic site of the G6PD locus to the X chromosome. The second study subjects a set of isochromosomal lines to sequential electrophoresis in an attempt to uncover common molecular heterogeneity within the global polymorphism, assuming that this variation may have gone undetected under conventional electrophoretic conditions. The genetic site was mapped following the segregation of the two common electrophoretic alleles, a so-called null allele, and two rare electrophoretic variants. From the pooled results, the Zw locus mapped to 62.9 on the X chromosome relative to the flanking markers car (at 62.5) and sw (at 64.7). A set of 126 iso-X chromosomal lines of diverse geographic origin was subjected to sequential electrophoresis under three different acrylamide conditions in addition to the conventional starch electrophoretic system. No additional variation beyond the common diallele polymorphism was seen.
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  • 82
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    Behavior genetics 10 (1980), S. 401-407 
    ISSN: 1573-3297
    Keywords: sexual isolation ; Drosophila ; geographic distance ; isolation index ; resource utilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract Six strains of the cosmopolitan speciesD. immigrans from the Australian life zone plus one from the USA, show weak sexual isolation and more rarely sexual selection. Levels of sexual isolation cannot be related to geographic distances. Assortative mating may have evolved as a byproduct of ecological divergence.
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    Behavior genetics 13 (1983), S. 17-27 
    ISSN: 1573-3297
    Keywords: pupation site ; pupation height ; artificial selection ; Drosophila ; density-dependent behavior ; genotype-environment interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract Selection for increased pupation height was carried out for 17 generations in two lines ofDrosophila simulans derived from a genetically heterogeneous base population. The realized heritability for mean pupation height in each line, calculated over the 17 generations, did not differ significantly from zero. Both selected lines tended to pupate away from the center of the culture medium to a greater extent than the control in the latter generations of the experiment but not in earlier generations. Pupation height may have been refractory to artificial selection because of an adaptation of this species to pupate on the larval food source. In a subsequent experiment, each line was tested at three larval densities in an apparatus different from the one used for selection. Each successively higher density showed a corresponding increase in pupation height. Both selected lines had higher mean pupation heights than the control line. The differences between lines became more pronounced as the larval density increased.
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    Biochemical genetics 21 (1983), S. 365-374 
    ISSN: 1573-4927
    Keywords: alcohol dehydrogenase ; NAD+ levels ; Drosophila
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Previous studies carried out in mammalian systems indicated that an organism's NAD+/NADH balance is carefully regulated but can be destabilized by dietary stresses. Since Drosophila alcohol dehydrogenase (ADH) uses NAD+ to remove a hydrogen from ethanol in the first step of alcohol catabolism, it is possible that under alcohol stress conditions the in vivo NAD+ levels in Drosophila may decrease. In this study genetically homozygous flies were stressed with maximally sublethal concentrations of ethanol (10%) for periods of up to 24 hr. The results indicate that NAD+ levels do in fact drop by at least 20% in response to ethanol stress. Evidence is presented that suggests that this decrease is the direct result of ADH-mediated catabolism.
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    Behavior genetics 22 (1992), S. 469-487 
    ISSN: 1573-3297
    Keywords: courtship ; pheromones ; Drosophila ; apterous ; juvenile hormone ; reproductive development ; sexual behavior
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract Theapterous (ap) gene ofDrosophila melanogaster exhibits extreme pleiotrophy: its functioning is essential for life, normal wing structure, juvenile hormone production, female fertility, and normal development of female sexual receptivity. Four mutantap alleles (ap 4,ap 56f,ap c, andap blt) were characterized for three additional phenotypes: male mating success, courtship behavior, and immature male sex appeal (the ability of males to stimulate homosexual cortship). Mating success with mature wild-type virgin females is reduced in males mutant for theap gene, the extreme case beingap 4/ap 4 males, which are behaviorally sterile. Inap mutants, nonwing courtship elements are qualitatively like those ofap +/ap + males. However, the mean rate of nonwing courtship directed toward virgin wild-type females (i.e., the mean temporal frequency of these displays) is reduced in males homozygous forap 4,ap 56f, orap c alleles. In contrast, theap blt allele makes for wild-type rates of nonwing courtship. Immature male sex appeal persists for at least 3 days in males homozygous forap c and, to a lesser extent, inap 56f orap 4 homozygotes;ap blt/ap blt and wild-type males lose immature male sex appeal after 1 day. All three male phenotypes map to theap locus, which is therefore essential for the development of normal levels of male courtship and male mating success and for the timely loss of immature male sex appeal. For each phenotype,ap + is dominant toap alleles making for behavioral abnormalities, with a single exception (for rate of nonwing courtship,ap +/ap c was low). For mating success and frequency of nonwing courtship, each allele pair exhibits at least partial complementation, except forap 4 andap 56f, which fail to complement. For immature male sex appeal,ap c,ap 4, andap 56f fall into the same complementation group. Juvenile hormone production is not correlated with effects on male reproductive behavior.
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    Behavior genetics 22 (1992), S. 557-573 
    ISSN: 1573-3297
    Keywords: Drosophila ; biometrical analysis ; behavior genetics ; genetic analysis ; ss a ; deletion mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract The homeotic mutationspineless-aristapedia (ss a ) transforms the aristae into second tarsi. Flies with aSS a phenotype also show extremely positive geotaxis as measured in a Hirsch-type geotaxis maze. Other antennal mutants and flies with their aristae amputated do not show such extreme positive geotaxis. Deletion analysis has comapped the geotaxis effect withSS a in band 89C on the third chromosome. Finally, a biometrical analysis has detected additional genes on the X chromosome that also affects geotaxis.
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    Journal of fluorescence 2 (1992), S. 47-62 
    ISSN: 1573-4994
    Keywords: Calcium ; fluorescence lifetime imaging ; calcium imaging ; fluorescence microscopy ; ratiometric imaging ; long-wavelength probes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract We describe imaging of calcium concentrations using the long-wavelength Ca2+ indicators, Calcium Green, Orange, and Crimson. The lifetimes of these probes were measured using the frequency-domain method and were found to increase from 50% to severalfold in response to calcium. The two-dimensional images of the calcium concentration were obtained using a new apparatus for fluorescence lifetime imaging (FLIM). We also describe procedures to correct for the position-dependent frequency response of the gain-modulated image intensifier used in the FLIM apparatus. Importantly, the FLIM method does not require the probe to display shifts in the excitation or emission spectra. Using the FLIM method, calcium imaging is possible using probes which display changes in lifetime in response to calcium. Consequently, calcium imaging is possible with excitation wavelengths ranging from 488 to as long as 620 nm, where autofluorescence and/or photochemical damage is minimal. These probes are also suitable for calcium measurements of single cells using lifetime-based flow cytometry.
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  • 88
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    Biochemical genetics 30 (1992), S. 159-168 
    ISSN: 1573-4927
    Keywords: Drosophila ; indirect flight muscle ; tropomyosin ; polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We describe polymorphism in aDrosophila indirect flight muscle-specific tropomyosin isozyme, named TnH-34. Three variants of this protein differ in their mobilities as determined by 1-D and 2-D SDS-PAGE. Meiotic mapping places the polymorphism close to, if not within, the structural gene encoding this tropomyosin isozyme. The most likely site of the mutations is within a single C-terminal exon. Flight-testing of different genotypes reveals that this variation in TnH-34 does not affect flight ability. These results suggest that some sequence variation may be tolerated in this section of the protein and correlate with the variability of this protein in different insect species.
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  • 89
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    Biochemical genetics 30 (1992), S. 305-315 
    ISSN: 1573-4927
    Keywords: Drosophila ; diaphorase ; purification ; kinetics ; immunochemical characteristics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Diaphorase-1 and diaphorase-2 were isolated from twoDrosophila species,D. virilis andD. melanogaster, and purified by gel filtration, affinity chromatography, immunoaffinity chromatography, and ion-exchange chromatography. The molecular weights of both enzymes were the same in each species. The molecular weight of diaphorase-1 was the same under both denaturating and nondenaturating conditions, close to 60,000, indicating a monomeric structure. Sodium dodecyl sulfate (SDS) electrophoresis of the purified diaphorase-2 revealed the presence of a single protein band of 55,000 Da, while the molecular weight of the native enzyme was found to be 67,000. The two diaphorases were further characterized by their pH optima, isoelectric points, and kinetic parameters, and antibodies were raised in rabbits against the purified enzymes fromD. virilis. The antibodies showed no cross-reactions but recognized the corresponding diaphorases inD. melanogaster andD. novamexicana as well asD. virilis. The data obtained confirmed the hypothesis of an independent genetic control of diaphorase-1 and diaphorase-2 inDrosophila.
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  • 90
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    Biochemical genetics 30 (1992), S. 305-315 
    ISSN: 1573-4927
    Keywords: Drosophila ; diaphorase ; purification ; kinetics ; immunochemical characteristics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Diaphorase-1 and diaphorase-2 were isolated from twoDrosophila species,D. virilis andD. melanogaster, and purified by gel filtration, affinity chromatography, immunoaffinity chromatography, and ion-exchange chromatography. The molecular weights of both enzymes were the same in each species. The molecular weight of diaphorase-1 was the same under both denaturating and nondenaturating conditions, close to 60,000, indicating a monomeric structure. Sodium dodecyl sulfate (SDS) electrophoresis of the purified diaphorase-2 revealed the presence of a single protein band of 55,000 Da, while the molecular weight of the native enzyme was found to be 67,000. The two diaphorases were further characterized by their pH optima, isoelectric points, and kinetic parameters, and antibodies were raised in rabbits against the purified enzymes fromD. virilis. The antibodies showed no cross-reactions but recognized the corresponding diaphorases inD. melanogaster andD. novamexicana as well asD. virilis. The data obtained confirmed the hypothesis of an independent genetic control of diaphorase-1 and diaphorase-2 inDrosophila.
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  • 91
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    Biochemical genetics 30 (1992), S. 77-83 
    ISSN: 1573-4927
    Keywords: acetylcholinesterase ; insecticide ; resistance ; Drosophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Selection of field populations originating from several countries allowed us to isolate 13 strains ofDrosophila melanogaster resistant to parathion.In vitro studies of acetylcholinesterase inhibition by paraoxon have been carried out on purified enzymes: most of the resistant strains harbor an altered acetylcholinesterase. Enzymes with higher resistance levels have been characterized with respect to their cross-resistance toward several insecticides. The patterns obtained have permitted us to group them and to delineate four categories. The existence of four distinct types of protein suggests that several mutations of acetylcholinesterase are responsible for insecticide resistance inDrosophila.
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  • 92
    ISSN: 1573-4927
    Keywords: Drosophila ; glycerol-3-phosphate dehydrogenase ; restriction map ; duplication ; enzyme activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Restriction site variation in a 25-kb region including thesn-glycerol-3-phosphate dehydrogenase (Gpdh) locus has been assessed in 29 single femaleD. melanogaster lines from the Cardwell (Australia, QLD) population. TheGpdh locus was duplicated in about one-third of the lines, although the duplication was incomplete and lacked exons 1 and 2. There was no restriction site variation in the duplicated region. Three insertions were found in the gene region but only one affected GPDH activity. The lines with the duplication had higher levels of GPDH activity and protein amount than did nonduplicated lines. This effect was also observed in lines extracted from two other Australian populations. The duplication is shown to have a similar structure in each population investigated and is also present in populations from China and Africa. It is suggested that the effect of the duplication on GPDH activity, which might be due to structural factors affecting transcription at theGpdh locus, could account for the worldwide distribution of the duplication.
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  • 93
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    Biochemical genetics 30 (1992), S. 515-527 
    ISSN: 1573-4927
    Keywords: glutathioneS-transferase ; Drosophila ; cellular detoxification ; pesticide resistance ; insect metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have isolated aDrosophila gene,DmGST-2, that encodes glutathioneS-transferase, a homo- or heterodimeric enzyme thought to be involved in detoxification of xenobiotics, including known carcinogens. The encoded protein has a primary sequence that is more similar to mammalian placental and nematode GSTs than that of a previously describedDrosophila GST gene, herein referred to asDmGST-1. We provide a physical map of the gene and show that it specifies at least two mRNAs, measuring 1.9 and 1.6 kb, which differ only in the lengths of their 3′ untranslated regions. Both of the mRNAs are present during all developmental stages.In situ hybridization of theDmGST-2 gene to larval polytene chromosomes places it within the 53F subdivision of chromosome 2, and Southern blotting to chromosomal DNA indicates that the gene has no close relatives within theDrosophila genome. Our results make possible molecular genetic approaches for further elaborating the function of glutathioneS-transferases in insect development and physiology, in the metabolism of plant toxins, and in conferring insecticide resistance.
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  • 94
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    Journal of comparative physiology 170 (1992), S. 605-613 
    ISSN: 1432-1351
    Keywords: Phototransduction ; Sensitivity ; Facilitation ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Deeply dark adapted (1 h) photoreceptor cells of the honey bee drone show a light-induced enhancement of sensitivity (facilitation) as an aftereffect of illumination or in the presence of dim backgrounds. The Ca2+-dependency of this effect was studied: Reduction of extracellular Ca2+ to 0.1 mM decreases the sensitivity of a dark adapted cell, and the light-induced increase in sensitivity due to repetitive, dim, 20 ms test flashes is slower than in normal saline. After a sensitizing conditioning light, the sensitivity drops faster in low-calcium saline. The light-induced enhancement of sensitivity is mimicked by pressure injections of low amounts of Ca2+ (Ca2+/EGTA-buffers; 0.15 μM free Ca2+) into a dark adapted cell. Injection of EGTA alone decreases the sensitivity. Injection of a solution containing ca1 mM free Ca2+ sequentially decreases and later increases the sensitivity transiently. These results suggest a model in which a progressive increase in intracellular Ca2+ concentration by light first increases (facilitates), and, at higher concentrations, decreases (light adapts) the sensitivity of the cells. One possible site of action for this positive and negative feedback control of cell sensitivity by Ca2+ is the endoplasmic reticulum.
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  • 95
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    Theoretical and applied genetics 83 (1992), S. 821-826 
    ISSN: 1432-2242
    Keywords: Drosophila ; Balancers ; Inversions ; Translocations ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We used a screen for maternally generated late embryonic lethals as a new method for the isolation of inversions that are suitable for the balancing of mutations in Drosophila hydei. The recovery of several inversions by this method demonstrates that female meiosis in D. hydei apparently differs from meiosis in female D. melanogaster, since in D. hydei the defective chromosomes which are generated by a single crossing-over within a paracentric inversion can be recovered via the egg nucleus. In addition, the classic method of crossingover suppression was used in order to isolate more inversions and to improve the balancing capacities of inversions. We succeeded in constructing chromosomes that allow the balancing of mutations on nearly the whole genome of D. hydei. We discuss here whether or not this method is suited for application to other organisms.
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  • 96
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    Theoretical and applied genetics 65 (1983), S. 173-180 
    ISSN: 1432-2242
    Keywords: Disruptive selection ; Linkage disequilibrium ; Genetic variance ; Drosophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Theoretical predictions of changes in variance with disruptive selection have used models of infinitely many genes so the increase in variance was necessarily due to linkage disequilibrium. With small numbers of loci, the disequilibrium is shown still to comprise the major part of the changes in variance. In a replicated experiment with Drosophila melanogaster, disruptive selection was practised for three generations, and this was followed by 5 or 7 generations of random mating. The heritability, as estimated from regression of progeny on parent, rose from 37% to 68% on selection, and subsequently declined to 45% on random mating. Changes of variance can be interpreted invoking the build up of linkage disequilibrium during selection followed by its breakdown upon relaxation. The results agree well with those obtained from Monte Carlo simulation.
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  • 97
    ISSN: 1432-2242
    Keywords: Drosophila ; Enzyme polymorphisms ; Latitudinal clines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The octanol dehydrogenase (Odh) and acid phosphatase (Acph) loci of Drosophila melanogaster are each polymorphic for two electrophoretically detectable alleles. The frequencies of the Odh and Acph alleles have been analysed in populations sampled from up to a 40 ° latitudinal range in each of Australasia, North America and Europe/Asia. Odh S frequency is found to be significantly negatively associated with distance from the equator in all three zones. The relationship of Acph S frequency to distance from the equator is significant and negative in Australasia but neither significant nor consistent in sign in North America and Europe/Asia.
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  • 98
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    Theoretical and applied genetics 57 (1980), S. 257-266 
    ISSN: 1432-2242
    Keywords: Drosophila ; Ethanol ; Climatic races ; Desiccation ; Development times
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Adult tolerance of ethanol vapour in a closed system containing 12% ethanol in solution, decreases in a cline from southern to northern Australia. However a Darwin population is more tolerant than predicted from its latitude. Ethanol tolerance races in Australia have almost certainly evolved within the last 100–150 years, because of resource unavailability prior to that time. Within populations, variation among isofemale strains is lowest in the climatically extreme southern Melbourne (37°S) and northern Darwin and Melville I. (11–12°S) populations. This suggests low resource diversity within extreme populations compared with the climatically less extreme Brisbane (28°S) and especially Townsville (19°S) populations. For desiccation resistance, the population rankings are: Darwin Melbourne 〉 Townsville 〉 Brisbane Melville I. and for development time, rankings are similar: Darwin Melbourne 〈 Townsville 〈 Brisbane Melville I. Therefore resource utilization heterogeneity is greatest in populations not greatly stressed by desiccation and where development times are extended. In total therefore, the utilization of a diversity of resources is a feature of populations tending somewhat towards a K-strategy; this is emphasized by the relative heterogeneity among isofemale strains of these populations for desiccation resistance and to a lesser extent development times. The D. melanogaster gene pool can be viewed as made up of climate-associated races. Since the ethanol tolerances of adjacent (and climatically similar) Darwin and Melville I. are very different, resource utilization races may occur within climatic races. Such a mosaic of resource utilization races are more likely in climatically extreme than in optimal habitats.
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  • 99
    ISSN: 1432-0878
    Keywords: Pituitary ; Pars intermedia ; Calcium ; Deionized water ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cytological changes in the pars intermedia of the goldfish were investigated after adding calcium to deionized water (DW). In fish maintained in DW, the PAS-positive cells are highly stimulated in comparison to cells of fish kept in fresh water (FW). In DW supplemented with calcium at the same concentration as in FW (2 mM/l), the hyperactivity of the PAS-positive cells is prevented. When calcium ions are added 60 h before the animals are sacrificed, the PAS positive cells start to show signs of regression and their granules are stored: the release of the granular material appears to be suppressed by calcium. In the goldfish, the PAS-positive cells, homologous to a similar cell type in the eel, react only very weakly with the PAS technique. The name “calcium-sensitive cells” appears to be more appropriate in the goldfish for this particular cell type, secreting an unknown factor. This factor, different from the prolactin produced in the rostral pars distalis of the hypophysis, might be an equivalent of a “hypercalcin”.
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  • 100
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    Cell & tissue research 213 (1980), S. 9-20 
    ISSN: 1432-0878
    Keywords: Pinocytosis ; Calcium ; Chlorotetracycline ; Fluorescence microscopy ; Amoeba proteus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The dynamics of Ca++ during induced pinocytosis were studied in Amoeba proteus using chlorotetracycline (CTC). The fluorescence of the Ca++ -CTC-complex was monitored by an image intensification system, which has certain advantages over standard equipment: (1) Living cells are not subjected to the damaging influence of intensive microscopic illumination, (2) fluorescent probes are not bleached during observation, and (3) the rapid dynamics of Ca++-fluxes can be recorded using short exposure times. The results demonstrate the existence of Ca++ bound to intracellular and extracellular sites of the cell membrane complex in normal locomoting and pinocytoting Amoeba proteus. The application of cations inducing pinocytosis causes a rapid decrease in the external CTC-fluorescence probably due to a release of Ca++ from the mucous layer. The degree of fluorescence intensity is correlated with the capacity of pinocytotic channel formation, i.e., the fluorescence decreases as the number of channels increases. During the phase of vesiculation a distinct fluorescence mainly restricted to the basal region of the channels is observed. Intracellular Ca++ was detected in close vicinity to the plasma membrane after both microinjection and external application of CTC. The internal CTC-fluorescence is slightly decreased during the induction phase of pinocytosis. The observations are in good agreement with previous results on the localization of Ca++-binding sites at the plasma membrane of Amoeba proteus and demonstrate the important role of Ca++-fluxes for the process of pinocytosis.
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