bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cloning and sequence analysis of a cDNA encoding an androgen-dependent mouse seminal vesicle secretory protein
Authors: Guilbaud C , Simon A M , Veyssière G , Jean C 
Published: December 2008

ABSTRACT
We report the cloning and sequencing of a new cDNA sequence encoding a protein from the mouse seminal vesicle. An open reading frame of 297 nucleotides encoded a protein of 99 amino acids with a calculated molecular mass of 11·454 kDa. The first 21 amino acids constituted a signal peptide followed by 78 amino acids encoding the secreted protein. The cDNA sequence comprised a 3′ untranslated region of 226 bp and the polyadenylation signal AATAAA, 19 bp upstream from the poly(A)+ tail. A high degree of homology was found between this protein and members of the family of seminal vesicle secretory (SVS) proteins, especially rat SVS VI. Northern blot analysis indicated the presence of a 0·7 kb mRNA species in the mRNAs of seminal vesicle tissue. Castration resulted in a marked decrease in the level of the 0·7 kb mRNA encoding the protein, whereas administration of testosterone to castrated males restored the 0·7 kb mRNA.
Discrete splicing alternatives in mRNA encoding the extracellular domain of the testis FSH receptor in the normal and hypogonadal (hpg) mouse
Authors: O'Shaughnessy P J , Dudley K 
Published: December 2008

ABSTRACT
The structure of RNA encoding the mouse testis FSH receptor was studied using reverse transcription and the polymerase chain reaction. Four major bands were observed by ethidium bromide staining and by hybridization to an FSH-receptor cDNA probe. The largest of these bands was the expected size (779 bp) while the other bands were spaced approximately 70 bp apart. Using alternative primers, each of the products was shown to contain exons 1, 9 and 10. Exons 2-8 in the FSH receptor gene are between 68 and 77 bp in size, suggesting that these multiple products arise by alternate splicing of the region encoding the extracellular domain of the receptor. A similar pattern of splicing was observed in cDNA from the testes of hypogonadal mice, showing that this alternative splicing pattern is not gonadotrophin-dependent.
Angiotensin II receptor isoforms in the rat adrenal gland: studies with the selective subtype antagonists DuP 753 and CGP42112A
Authors: Montiel M , Barker S , Vinson G P , Jiménez E 
Published: December 2008

ABSTRACT
The angiotensin II (Ang II)-binding sites in rat adrenal gland membranes were characterized using 125I-radiolabelled Ang II. While Scatchard analysis identified a single population of Ang II receptor sites, isoelectric focusing (IEF) on polyacrylamide gels revealed four peaks of specific Ang II binding which migrated to isoelectric points (pI values) 6·8, 6·7, 6·5 and 6·3. In binding assays in the presence of an excess of the Ang II receptor AT1 subtype antagonist DuP 753, a monophasic dose-dependent displacement of 125I-labelled Ang II binding by the Ang II receptor AT2 subtype antagonist CGP42112A was observed, and vice versa. In this system, reduction of disulphide bridges using 1 mmol dithiothreitol (DTT)/l markedly increased the number of binding sites in the adrenal zona glomerulosa without affecting receptor affinity.
Using IEF, it was found that both DuP 753 and CGP42112A were able to reduce specific binding of each of the four peaks to some extent. However, the predominant effect of DuP 753 was to reduce the labelling of the isoform at pI 6·7 substantially, while CGP42112A significantly inhibited the specific 125I-labelled Ang II binding to the pI 6·3 isoform. When DuP 753 and CGP42112A were used together, specific binding of 125I-labelled Ang II to the isoforms of pI values 6·8, 6·7 and 6·3 was completely eliminated. These data suggest that the four peaks of specific binding found may be composed of different isoforms of both AT1 and AT2 receptor subtypes and that the Ang II receptor isoforms which migrated to pI 6·7 and pI 6·3 are predominantly composed of AT1 and AT2 receptor subtypes respectively. Interestingly, in the presence of both antagonists, 8·7 ± 0·9% of the specific binding migrating at pI 6·5 remained unaffected. This finding suggests the presence of an additional subtype, which is neither AT1 nor AT2, in the rat adrenal zona glomerulosa.
In further studies, pretreatment with DTT was found to increase the specific 125I-labelled Ang II binding of all four isoforms. Moreover, DTT also produced a further specific binding component between pI 6·5 and pI 6·7 which exhibited AT2 subtype pharmacology in DTT-treated preparations. Since DTT has been reported to enhance only AT2 subtype binding this also suggests that the different isoforms may contain components related to both AT1 and AT2 receptor subtypes.
Developmental regulation of bovine insulin-like growth factor-II (IGF-II) gene expression: homology between bovine transcripts and human IGF-II exons
Authors: Boulle N , Schneid H , Listrat A , Holthuizen P , Binoux M , Groyer A 
Published: December 2008

ABSTRACT
Initial observations have indicated similarities between bovine and human IGF-II production during development. The aim of the present study was to investigate whether cattle could provide an experimental model that would mimic the complex pattern of human IGF-II gene expression. Expression of bovine IGF-II gene during development was studied by RNA hybridization using various human IGF-II probes. In fetal tissues and in adult muscle, the bovine IGF-II gene was expressed as a family of eight transcripts ranging in size from 5·2 to 1·1 kb. In adult bovine liver, a major IGF-II transcript of 4·4 kb was expressed that could not be detected in any fetal or adult extra-hepatic tissue. During fetal life, quantitative IGF-II mRNA expression differed in liver and muscle, and the relative amounts of the different transcripts varied with the tissue of origin. These observations suggest that the regulation of bovine IGF-II gene expression is specific to the stage of development and the tissue concerned. Moreover its pattern is very similar to that in its human counterpart.
In order to identify a putative homology between human and bovine gene structures, bovine mRNAs were examined for cross-hybridization with various non-coding exons of the human gene. Cross-hybridization was detected with human untranslated exons 5 and 6, suggesting the presence of two distinct promoters similar to the human promoters P3 and P4. The 4·4 kb mRNA species expressed in adult bovine liver failed to hybridize to a probe for human exons 1 and 2, suggesting that the leader sequences of this transcript were different from those present in the human gene. Finally, results obtained with a probe containing the 3′ untranslated end of exon 9 suggested the presence of at least two polyadenylation sites in the bovine gene.
Although differences in IGF-II gene structures were found between cattle and man, the similarities in the pattern of gene expression between the two species suggest that cattle may be a useful model to investigate some developmental aspects of the expression of the human IGF-II gene.
Transferrin gene expression in the mammary gland of the rat. The enhancing effect of 17β-oestradiol on the level of RNA is tissue-specific
Authors: Escalante R , Houdebine L-M , Pamblanco M 
Published: December 2008

ABSTRACT
We have investigated the physiological factors which regulate transferrin gene expression in the mammary gland of the rat. Our studies by dot blot analysis have demonstrated that multiple doses of 17β-oestradiol (OE2; 0·5 mg/kg per day for 3 days) elicit a specific 3·5-fold increase in the transferrin mRNA levels in the mammary glands of virgin rats. The hormonal action of OE2 in mammary tissue was specific for the transferrin gene, as judged by hybridization with β-actin cDNA. The accmulation of transferrin mRNA induced by OE2 treatment was similar to the developmentally regulated expression of the gene observed during the reproductive cycle. The steady-state level of mammary transferrin mRNA increased by up to 4·5-fold at day 21 of lactation, when compared with virgin and pregnant rats. Our results show that the pattern of transferrin gene expression is different in mouse and rat mammary glands. The specific response of the transferrin gene to OE2 was not found in the liver or in the uterus. In the uterus alone, OE2 produced a significant increase in the content of nucleic acids and also induced the accumulation of transferrin and β-actin mRNAs. We have detected for the first time an induction of transferrin gene expression in the mammary gland in response to OE2, and these results support the view that the pattern of transferrin gene multimodulated expression is tissue- and species-specific.
Expression of growth hormone genes in Atlantic salmon
Authors: Lorens J B , Nerland A H , Aasland R , Lossius I , Male R 
Published: December 2008

ABSTRACT
Atlantic salmon (Salmo salar) possess two genes encoding GH. We have investigated the expression of these two genes in the salmon pituitary. The transcriptional start site was localized 64 nucleotides upstream of the first methionyl codon using primer extension and 5′ specific polymerase chain reaction (PCR) assays. Northern analysis revealed a major Atlantic salmon GH (salGH) transcript band of approximately 1400 nucleotides. As coexpression of the salGH genes is not discernible by transcript length, other techniques were used to assess gene activity; RNase protection analysis revealed GH transcript heterogeneity, while reverse transcription-PCR assays detected transcripts from both genes at approximately equivalent amounts. The encoded salGH protein, generated in vitro and by Escherichia coli, shares electrophoretic and immunoreactive identity with native pituitary salGH.
Detection of gastrin mRNA in fresh human colonic carcinomas by reverse transcription-polymerase chain reaction
Authors: Monges G , Biagini P , Cantaloube J-F , Chicheportiche C , Frances V , Brandini D , Parc P , Seitz J-F , Giovannini M , Sauvan R , Hassoun J 
Published: December 2008

ABSTRACT
To investigate the hypothesis that gastrin might be synthesized by tumour tissues in cancer of the colon, samples from six human colon tumours, one hepatic metastasis, four normal colonic mucosal samples and two antral and one fundic gastric mucosal samples from nine patients were analysed to determine whether gastrin mRNA was present.
RNA was extracted from surgical specimens by ultracentrifugation on a CsC1 cushion, purified using the guanidinium thiocyanate method, reversetranscribed and amplified by polymerase chain reaction. Gastrin mRNA was detected in each colonic carcinoma sample (including the hepatic metastasis), while no such signal was observed in normal colon biopsies. Positive and negative controls (gastric antrum and fundus respectively) gave the expected results. In each of the positive samples, the chemiluminescent revelation of amplified products after Southern blotting corresponded to gastrin mRNA without the intron.
These findings demonstrate the ability of primary and metastatic human colonic tumours to produce gastrin mRNA. Since malignant cell lines have been reported to produce gastrin peptide, and since gastrin receptors were present in some cases, our results support the idea that gastrin may be involved in an autocrine mechanism.
Influence of growth factors on the plasminogen activator activity of avian granulosa cells from follicles at different maturational stages of preovulatory development
Authors: Lafrance M , Croze F , Tsang B K 
Published: December 2008

ABSTRACT
Granulosa cells from the first (Fl), third (F3) and fifth and sixth (F5–6) preovulatory follicles and the small yellow follicles (SYFs; diameter 6–8 mm) were cultured for 21 h in the absence and presence of murine and human epidermal growth factors, fibroblast growth factor, transforming growth factors α and β-I (TGFα, TGFβ), platelet-derived growth factor and insulin-like growth factor-I at concentrations of 0·1–100 ng/ml. Plasminogen activator (PA) activities in the cell (PAc) and in the medium (PAm) were measured by fibrinolysis and fibrin overlay methods. Basal PAc and PAm activities were highest in cell cultures from the less mature follicles (F5–6 and SYF) and decreased as the follicles matured (F3>F1). PAc activity was greater than PAm activity, irrespective of the stage of follicular development. All growth factors examined at the 100 ng/ml level were effective in increasing PAc and PAm activities in cultures of granulosa cells from Fl follicles. However, only TGFα was able to increase PA activities at lower concentrations. The stimulation of the PA activities of granulosa cells from F3 follicles was inconsistent. None of the growth factors significantly increased PA activities in granulosa cells from F5–6 follicles and SYFs, as determined by fibrinolysis. The major PAc and PAm species (characterized by fibrin overlay) had a molecular mass of about 35 kDa, which is characteristic of the urokinase type. Both assay methods detected a stimulatory effect of the growth factors on PA activities in the granulosa cells from Fl follicles. However, an increase in PA activities in cells from F3 and F5–6 follicles and SYFs was indicated only after fibrin overlay analysis. Tritiated thymidine was incorporated into the DNA of granulosa cells at all stages of follicular development and was enhanced by all growth factors, although TGFα and TGFβ were the most effective and had a ranked order of activity: F3, F5–6>F1, SYF. The present findings show that, of the growth factors examined, TGFα may be an effective regulator of PA activity in avian granulosa cells during follicular development, in addition to its observed mitogenic action.
Evidence that two tilapia (Oreochromis niloticus) prolactins have different osmoregulatory functions during adaptation to a hyperosmotic environment
Authors: Auperin B , Rentier-Delrue F , Martial J A , Prunet P 
Published: December 2008

ABSTRACT
Two forms of prolactin (tiPRLI and tiPRLII), with only 69% sequence identity, have been previously described in the cichlid fish tilapia (Oreochromis species). In the present study we have attempted to investigate the biological activity of these two prolactin forms during adaptation to a hyperosmotic environment. For this purpose, we have developed two highly sensitive (sensitivity: 0·05 ng/ml) and specific (cross-reactivity <0·04%) radioimmunoassays for tiPRLI and tiPRLII, using recombinant hormones.
When fish were directly transferred from fresh to brackish water, the measured levels of plasma tiPRLI and tiPRLII dropped abruptly until 12 h after transfer. Thereafter, plasma tiPRLII remained stable (around 0·5 ng/ml) until the end of the experiment, whereas plasma tiPRLI continued to decrease to undetectable levels. These different patterns of change are reflected in the calculated ratio of plasma tiPRLII to tiPRLI, which increased from 2–3 in fresh water-adapted fish to over 10 in fish which had spent 3 days or more in brackish water. The pituitary contents of tiPRLI and tiPRLII varied in a qualitatively similar fashion after transfer to brackish water. The tiPRLI content dropped continuously after 12 h, reaching one-twelfth of its initial level after 2 weeks. The pituitary tiPRLII content, on the other hand, did not decrease significantly until day 7, and after a 2-week exposure to brackish water it had only decreased by 50%.
When injected into tilapia adapted to brackish water, both ovine prolactin and recombinant tiPRLI induced a clear dose-dependent ion-retaining effect. In contrast, the effect induced by tiPRLII treatment was markedly smaller and not dose-dependent.
Northern blot analysis of tiPRL mRNAs using either a tiPRLI or a tiPRLII cDNA probe indicated the presence of two mRNAs differing in size: a 1·7 kb mRNA coding for tiPRLI and a 1·3 kb mRNA coding for tiPRLII. After transfer to brackish water, levels of the two mRNAs decreased similarly.
The present study indicates that, in O. niloticus, the two forms of prolactin have different osmoregulatory roles during adaptation to brackish water. Accordingly, their syntheses are differentially regulated after transfer to a hyperosmotic environment, presumably at a post-transcriptional level.
Induction of gene expression during involution of the lactating mammary gland of the rat
Authors: Guenette R S , Corbeil H B , Léger J , Wong K , Mézl V , Mooibroek M , Tenniswood M 
Published: December 2008

ABSTRACT
After weaning, the mammary gland ceases lactation and involutes. The wet weight of the gland decreases by 70% within 4 days of weaning. This involves significant tissue remodelling as the ducts regress and return to the resting state. The presence of apoptotic bodies in the luminal epithelial compartment 2 to 3 days after weaning provides clear evidence that a substantial proportion of the regression is attributable to the induction of active cell death (ACD) of the epithelial cells.
These changes in the architecture of the gland were found to be mirrored by changes in gene expression. The steady-state level of β-casein mRNA decreased rapidly after weaning from the high levels seen during lactation to undetectable levels by 8 days after weaning. The steady-state levels of expression of a number of genes associated with ACD, including TRPM-2, tissue transglutaminase (TGase) and poly(ADP-ribose) polymerase (PARP), increased transiently during this time-frame. The steady-state level of TRPM-2 mRNA increased 2 days after weaning, reaching a peak on day 4, and decreasing to undetectable levels by day 8 after weaning. The steady-state levels of two other mRNAs, TGase and PARP, showed very similar kinetics. In contrast, the mRNA for Hsp 27, which has been shown to be induced during prostate regression, was not significantly induced in the regressing mammary gland. In-situ hybridization demonstrated that the TRPM-2, TGase and PARP genes were expressed predominantly in the luminal epithelial cells of the ducts. These cells expressed β-casein mRNA during lactation, and underwent ACD after weaning.
While the ultrastructural changes in the mammary gland after weaning, and the induction of TRPM-2, TGase and PARP mRNAs, are reminiscent of apoptosis in the prostate, several features of the process are different. Most notably, the disruption of the secretory processes and the lack of increased expression of Hsp 27 in the regressing mammary gland suggest that there may be a number of important events in ACD that are not common to all cells.
Mutation in 3ß-hydroxysteroid dehydrogenase type II associated with pseudohermaphroditism in males and premature pubarche or cryptic expression in females
Authors: Mendonça B B , Russell A J , Vasconcelos-Leite M , Arnhold IJ P , Bloise W , Wajchenberg B L , Nicolau W , Sutcliffe R G , Wallace A M 
Published: December 2008

ABSTRACT
A mutation (A82T) is described in the coding sequence of the gene for 3ß-hydroxysteroid dehydrogenase (3ß-HSD) type II that is associated with variable clinical consequences. Four homozygotes are described, all of which showed elevated levels of Δ5 steroids consistent with 3ß-HSD deficiency. Two males from a consanguineous family were found to be homozygous for A82T and were affected with pseudohermaphroditism. They differed in their degree of mild salt loss. In the same family a female was found to be homozygous for A82T, but was clinically normal and had no history of premature pubarche or of abnormal menstrual cycles. However, in an apparently unrelated family, the A82T mutation was found in a female affected with premature pubarche. This is the first report of a proven mutation in 3ß-HSD type II associated with premature pubarche.
Ontogeny of steroidogenic enzyme expression in the porcine conceptus
Authors: Conley A J , Rainey W E , Mason J I 
Published: December 2008

ABSTRACT
This study examined fetal steroidogenic enzyme expression and function during pregnancy in the pig. Northern and Western analyses were performed to detect the cytochrome P450 enzyme 17α-hydroxylase/17–20 lyase (P450c17) and that for cholesterol side-chain cleavage (P450scc), as well as 3β-hydroxysteroid dehydrogenase (3β-HSD) expression in several porcine fetal tissues. The data demonstrate higher steroidogenic enzyme expression in the fetal adrenal glands and testes than in the placenta at all stages of development examined. Although steroidogenic enzyme expression was maintained throughout gestation in both the fetal adrenals and the testes, adrenal P450c17 expression was higher in the early and late stages when compared with the intermediate stages of fetal development. The stimulation of fetal adrenal steroidogenic enzyme expression in the later stage fetuses was accompanied by increased expression of P450c17 in both the fetal testes and placenta. The expression of 3β-HSD by porcine fetal testes was low compared with that of the fetal adrenal gland at all stages of development. Adrenal explants and cultured cells secreted cortisol and androstenedione but much lower amounts of corticosterone, dehydroepiandrosterone and aldosterone. Secretion of cortisol and androstenedione by adrenal explants was maintained by ACTH for 5 days of culture but declined in controls. In cultured porcine fetal adrenal cells, ACTH and angiotensin II stimulated the secretion of multiple steroids. Porcine fetal testis explants and cultured cells secreted testosterone, dehydroepiandrosterone and androstenedione, but were only moderately responsive to trophic stimulation by LH. In general, the data suggest that the fetal adrenal glands and the fetal testes have the potential to contribute significantly to the production of steroids during pregnancy in pigs.
Differential regulation of c-fos gene expression by two types of human endothelin receptor in Chinese hamster ovary cells
Authors: Tabuchi H , Furuichi Y , Miyamoto C 
Published: December 2008

ABSTRACT
To investigate the nuclear signalling pathway induced by endothelin (ET) isopeptides, we have established permanent Chinese hamster ovary (CHO) cell lines, CHO-ETA/fos-lacZ and CHO-ETB/fos-lacZ, that produce both a c-fos-β-galactosidase fusion protein and either the type A or the type B human ET receptor. These cell lines permitted a colorimetric measurement of c-fos expression, which was induced by the signal transduction system with ET receptors and ET isopeptides. We found that the ET-1-dependent c-fos expression was so efficient that it could respond to low concentrations (even a physiological concentration) of ET-1. For example, CHO-ETA/fos-lacZ and CHO-ETB/fos-lacZ responded to ET concentrations of 5×10−9m and 5×10−13m respectively. Using this highly sensitive system, the H-7 sensitive protein kinase was found to be involved in signal transduction mediated by ETA, and also partly in the ETB-mediated pathway. These lines of evidence suggest that c-fos expression occurs through at least two different pathways, depending on the concentration of ET in plasma.