bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of excitatory amino acid pathways in the control of pituitary function in neonatally oestrogenized male rats
Authors: Pinilla L , Tena-Sempere M , Aguilar E 
Published: December 2008

Abstract
It is well known that in the rat neonatal manipulation of the sex steroid environment results in altered hypothalamic-pituitary function in adulthood which implies an abnormal prolactin secretion and gonadotrophin response to orchidectomy. The present paper analyses the involvement of excitatory amino acid pathways in the disturbed gonadotrophin and prolactin secretion in male rats neonatally injected with oestradiol benzoate (500 μg on the day of birth). In the first experiment, 60-day-old control and oestrogenized male rats (intact or orchidectomized a week before) were killed 15 min after injection of vehicle, N-methyl-d-aspartic acid (NMDA; 15 mg/kg) or kainic acid (KA; 15 mg/kg). In the second experiment, prepubertal males were killed 15 min after injection of vehicle or NMDA. In the third experiment 30-, 45-, 60-and 90-day-old intact control and oestrogenized males were killed 15 min after injection of vehicle or KA. In the fourth experiment, control and oestrogenized males were sham-orchidectomized, orchidectomized or orchidectomized and implanted with Silastic capsules at 30 days of age and killed on day 90, 15 min after vehicle or KA injection.
We found that (1) as previously reported, oestrogenized males showed hyperprolactinaemia and absence of LH and FSH responses after orchidectomy, (2) the stimulatory effect of NMDA on LH secretion was observed in prepubertal control and in prepubertal and adult oestrogenized males, (3) KA stimulated LH secretion only in prepubertal (30-day-old) and peripubertal (45-day-old) control males, whereas stimulation was observed in oestrogenized males at all ages studied, except when they were implanted with testosterone, (4) FSH secretion was only stimulated in oestrogenized orchidectomized males, and (5) prolactin secretion was inhibited by NMDA and KA in control and oestrogenized males.
The greater effectiveness of NMDA and KA on gonadotrophin secretion in oestrogenized males, a finding especially noticeable after orchidectomy, may reflect the existence of a greater pool of releasable GnRH, a consequence of the absence of response to orchidectomy. The decrease in prolactin secretion after NMDA or KA administration in oestrogenized males may be mediated by an increase in dopaminergic tone, presumably reduced in the hyperprolactinaemic oestrogenized males. Finally, the effectiveness of KA on prolactin and LH secretion was maintained in adult oestrogenized males, perhaps because of the permanent decrease in testosterone secretion.
Journal of Endocrinology (1995) 147, 51–57
Interaction between activin A and cAMP in the induction of FSH receptor in cultured rat granulosa cells
Authors: Nakamura M , Nakamura K , Igarashi S , Tano M , Miyamoto K , Ibuki Y , Minegishi T 
Published: December 2008

Abstract
The acquisition of FSH receptor during preantral folliculogenesis is believed to be a key step in the subsequent development of follicles. We examined the interaction between activin and cAMP in FSH receptor induction in rat granulosa cells by measuring 125I-FSH binding and FSH receptor mRNA. In the 125I-FSH binding study, 0·2 mm 8-Br-cAMP and 1 μm forskolin were maximally effective in FSH receptor induction (169 and 220% respectively of control), while higher concentrations gave attenuated responses. It appears that cAMP has ambivalent effects on FSH receptor induction depending on the concentration and length of exposure. Activin alone dramatically increased the number of FSH receptors (314% of control). Moreover, synergistic effects of activin and 8-Br-cAMP or forskolin were observed on FSH receptor induction: a combination of activin (80 ng/ml) and low doses of 8-Br-cAMP (0·2 mm) or forskolin (1 μm) was most effective (160 or 140% of that induced by activin alone) and receptor levels reached a maximum at 24 h. These levels then markedly decreased after 72 h of incubation. Northern blot analysis revealed that the combination of activin (80 ng/ml) and 8-Br-cAMP (0·2 mm) or forskolin (1 μm) increased FSH receptor mRNA to about 140% of that induced by activin alone. These results indicate that activin and cAMP induced FSH receptor synergistically. However, activin did not enhance the production of cAMP induced by forskolin. In addition, a protein kinase A inhibitor (H89) (2 μm), which inhibited the effects of forskolin, had no effect on the action of activin. Taken together, the present findings suggest that the action of activin is not via a cAMP pathway, and that activin works co-operatively with cAMP on folliculogenesis.
Journal of Endocrinology (1995) 147, 103–110
Induced luteal regression in the primate: evidence for apoptosis and changes in c-myc protein
Authors: Fraser H M , Lunn S F , Cowen G M , Illingworth P J 
Published: December 2008

Abstract
There is increasing molecular evidence that apoptosis is involved in the process of structural luteal regression in non-primate species. Apoptosis is dependent upon the activation of certain proto-oncogenes and c-myc protein has an important regulatory role in this process in some cell types. The aim of the present study was to determine the occurrence and localisation of c-myc protein within the primate corpus luteum, determine changes during induction of luteal regression and examine the corpora lutea for morphological evidence of apoptosis. Ovaries were studied from marmoset monkeys in the late follicular, and in the early, mid and late luteal phases. Luteal regression was induced either by treatment with prostaglandin F2α analogue or GnRH antagonist administered during the mid luteal phase and ovaries obtained 24 and 48 h later. Immunocytochemistry was performed using a monoclonal antibody to the c-myc protein. In pre-ovulatory follicles positive staining was found in the nucleus of a few granulosal cells and in the cytoplasm of thecal cells. c-myc was present in all corpora lutea where it was localised predominantly in the cytoplasm. In early corpora lutea, scattered cells with intense staining were observed in the presence of a majority of moderately or weakly stained cells. In the mid and late luteal phases, corpora lutea were uniformly moderately stained for c-myc. Following induction of luteal regression, nuclear degeneration with condensation and fragmentation indicative of apoptosis was observed. In other luteal cells, increased cytoplasmic volume and dissolution of cellular and nuclear membranes suggested necrosis. After luteal regression the overall intensity of staining for c-myc declined, but was present at high signal concentration in the cytoplasm of those cells whose morphological integrity was best maintained following treatment. In a minority of steroidogenic luteal cells, both nuclear and cytoplasmic staining was observed. These results suggest that after ovulation there appears to be a marked increase in c-myc production in the cytoplasm of the luteal cells of the developing corpus luteum and that c-myc is present throughout the luteal phase. During induced luteal regression c-myc may undergo a transitory rise and transfer to the nucleus and both apoptosis and necrosis occur during the process of luteolysis.
Journal of Endocrinology (1995) 147, 131–137
Charge heterogeneity of the AT1 angiotensin II receptor subtype in the rat lung
Authors: Montiel M , Quesada J , Jiménez E 
Published: December 2008

Abstract
In order to obtain more information on the molecular structure of the angiotensin II (Ang II) binding sites from whole rat lung membranes these were characterized by isoelectric focusing (IEF) and SDS-PAGE. Whereas a single population of Ang II receptor sites was identified (Kd=2·2± 0·3 nmol/l; Bmax=203·9± 15·8 fmol/mg protein) by Scatchard analysis, using IEF three Ang II binding isoforms were observed; a major band which migrated to isoelectric point (pI) 6·7, and two minor bands with pI values of 6·5 and 6·3 Specific binding of 125I-Ang II to rat lung membrane preparations was sensitive to Losartan, a non-peptide AT1, receptor subtype antagonist, but was unaffected by the AT2 receptor subtype antagonist CGP42112A. Immunoblotting analyses on SDS gels, using a monoclonal antibody specific to the AT1, receptor, showed two immunoreactive protein species of 45 and 48 kDa. Enzymic deglycosylation using recombinant N-glycanase did not alter the molecular weight patterns of the AT1, receptor subtype. The results of the present study demonstrated that the Ang II receptor population in the whole rat lung consists solely of the AT1, receptor subtype and that the AT2 receptor subtype is absent. In addition, the data showed the existence of charge heterogeneity of the AT1, receptor subtype, and suggest that glycosylation probably does not contribute to its charge heterogeneity.
Journal of Endocrinology (1995) 147, 153–159
Functional luteolysis in response to hydrogen peroxide in human luteal cells
Authors: Vega M , Carrasco I , Castillo T , Troncoso J L , Videla L A , Devoto L 
Published: December 2008

Abstract
To evaluate the effect of reactive oxygen species in human corpus luteum function, we investigated whether hydrogen peroxide (H2O2) affects the in vitro luteal cell production of steroids. H2O2 treatment (1·0–100 μm) of mid and late luteal cell cultures elicited a dose-dependent decrease in basal progesterone production. However, treatment of mid luteal cells with a low concentration of H2O2 0·01 μm) significantly stimulated progesterone secretion (P<0·05). In addition, H2O2 (100 μm) markedly inhibited human chorionic gonadotropin (hCG)-stimulated progesterone and estradiol secretion. cAMP production was enhanced (2·4-fold, P<0·05) by hCG treatment of luteal cells. The addition of H2O2 (0·1–100 μm) to hCG-stimulated luteal cell cultures elicited a decrease in cAMP concentration (P<0·05) and in the specific binding of radiolabeled hCG by luteal cells. Progesterone and estradiol production stimulated by dibutyryl cAMP were significantly inhibited by H2O2 (P<0·05). These findings suggest that H2O2 interferes with basal steroid production and, in hCG-stimulated conditions, it may inactivate the gonadotropin–receptor complex. The anti-steroidogenic action of H2O2 therefore raises the possibility of a modulatory role of H2O2 in human luteal steroidogenesis.
Journal of Endocrinology (1995) 147, 177–182
Pituitary and plasma levels of growth hormone in Booroola sheep that are either homozygous carriers or non-carriers of the FecBB fecundity gene
Authors: Moore L G , McNatty K P , Isaacs K L , Lun S , Ng Chie W , McNatty S , Hudson N L 
Published: December 2008

Abstract
The aim of this study was to examine the effect of the FecBB fecundity gene on plasma concentrations and pituitary content of growth hormone (GH) in sheep. No differences were found between homozygous carriers (BB) and non carriers (++) of the FecBB gene with regard to pituitary GH contents in both ovariectomized and intact ewes. However, ovariectomized ewes had higher levels of pituitary GH than intact ewes (P<0·01). There were no differences between FecBB genotypes with respect to plasma concentrations of GH in 6-year-old ovariectomized ewes bled every 10 min for 12 h or in ram lambs bled weekly during their first year of life. GH levels in the rams decreased until week 27, increased to a peak at week 31 then decreased before increasing again at week 43. Mean plasma GH concentrations in the ewe lambs bled weekly for a year decreased until week 19 then remained at approximately this level for the remainder of the year. Mean GH plasma concentrations in the ram lambs were higher than in the ewe lambs (P<0·001). Ewe lambs that were homozygous for the FecBB gene had lower body weights (P<0·05) and had higher levels of GH (P<0·01) than non carrier ewe lambs during their first year. Before the average age of first behavioural oestrus (36 weeks) GH levels in the ewe lambs were negatively correlated with body weights (r=−0·69, P<0·001, n=22). When body weight was included as a covariate in analysis of variance the genotype difference in ewe lamb plasma GH concentrations was no longer significant. In summary, pituitaries from ovariectomized ewes had higher levels of GH than those from intact ewes. There were no FecBB gene specific differences in pituitary levels of GH, the profile of plasma GH in 6-year-old ovariectomized ewes or in ram lambs during their first year of life. BB ewe lambs had higher levels of GH than ++ ewe lambs during their first year; however, this difference was probably due to the BB ewes having lower body weights than the ++ ewes because body weight was negatively correlated with mean GH levels.
Journal of Endocrinology (1995) 147, 217–223
Immunolocalisation of androgen receptor to interstitial cells in fetal rat testes and to mesenchymal and epithelial cells of associated ducts
Authors: Majdic G , Millar M R , Saunders P T K 
Published: December 2008

Abstract
Androgens are required for the development of male internal and external genitalia. Androgen action is mediated by an intracellular receptor which acts as a transcription factor following activation by ligand binding. The aim of the present study was to define the time of appearance of androgen receptor (AR) in the male fetal rat gonad using immunohistochemistry. Intact fetuses (days 13·5–16·5) or testicular tissue (days 16·5–20·5 and days 3–7 postnatal) were fixed in Bouins' solution and processed into paraffin wax. On day 16·5 nuclear AR were present in mesenchymal cells surrounding the Wolffian duct but those around the Mullerian duct were receptor negative. During the following day (17–18) the abundance of nuclear staining increased, becoming detectable in the epithelial cells of the Wolffian ducts. Within the testis some nuclear staining was apparent at day 17 but was confined to interstitial cells surrounding the seminiferous cords. As development of the testis proceeded the abundance of nuclear AR in peritubular and elongated mesenchymal cells increased. AR were not detected in fetal Leydig cells expressing 3β-hydroxysteroid dehydrogenase nor in the ovaries or associated ducts of female fetuses at the same ages.
In conclusion, in the rat we have found AR expression detectable by immunohistochemistry in mesonephric mesenchyme to be confined to that underlying the Wolffian ducts and to be absent from the area around the degenerating Mullerian duct. On and after day 17 of gestation AR is present in Wolffian duct epithelial cell nuclei and within the testis it is confined to peritubular and interstitial cells which may have migrated from the mesonephros. Fetal Leydig cells were receptor negative. Within the seminiferous cords AR in Sertoli cells remained low until after birth and some perinuclear staining was detected in cells thought to be gonocytes. We believe this to be the first report of immunolocalisation of AR to fetal testicular interstitial cells.
Journal of Endocrinology (1995) 147, 285–293
Differential expression of 11β-hydroxysteroid dehydrogenase 1 and 2 in the developing ovine fetal liver and kidney
Authors: Langlois D A , Matthews S G , Yu M , Yang K 
Published: December 2008

Abstract
In adult mammals, liver and kidney are the two major sites of biosynthesis for 11β-hydroxysteroid dehydrogenase (11β-HSD) 1 and 2 respectively. In the present study, the expression of these two isozymes in the developing ovine fetal liver and kidney was characterized. Livers and kidneys were obtained from fetal sheep at days 85, 100–120 and 140–143 of gestation (term=145 days). Tissue levels of 11β-HSD2 mRNA were assessed by Northern blot analysis. 11β-HSD dehydrogenase and reductase activities in tissue homogenates were determined by a radiometric conversion assay using cortisol and cortisone as physiological substrates respectively. The unidirectional 11β-HSD2 dehydrogenase activity was identified by its distinct co-factor preference (NAD), and by its unique ability to metabolize dexamethasone (Dex). In the liver, 11β-HSD1 dehydrogenase and reductase activities were present by day 85, and their levels did not change between days 85 and 100–120 but increased more than twofold at days 140–143. This was consistent with changes we reported previously in the fetal hepatic 11β-HSD1 mRNA. 11β-HSD1 reductase activity was always higher than the dehydrogenase activity. 11β-HSD2 mRNA and activity were undetectable in the fetal liver at all three ages. By contrast, 11β-HSD2 mRNA was present in the fetal kidney by day 85, and its abundance increased progressively thereafter. There was a parallel increase in the renal 11β-HSD2 activity. Dex was also converted to 11-dehydro-Dex by the fetal kidney. In keeping with the absence of the full-length 11β-HSD1 mRNA, 11β-HSD1 activity was undetectable in the kidney. These results indicate that (1) 11β-HSD1 and 2 genes are differentially expressed and regulated in the fetal liver and kidney during development, (2) since the hepatic 11β-HSD1 reductase activity is always higher than the dehydrogenase activity, the fetal liver may be a potential extra-adrenal source of cortisol, and (3) 11β-HSD2 in the kidney may play a very important role in protecting the fetus from elevated levels of bioactive glucocorticoids.
Journal of Endocrinology (1995) 147, 405–411
Adrenalectomy induces a decrease in the light scatter properties and amylase content of isolated zymogen granules from rat pancreas as analyzed by flow cytometry
Authors: Garcia-Montero A C , De Dios I , Rodriguez A I , Orfao A , Manso M A 
Published: December 2008

Abstract
The effect of glucocorticoid deprivation induced in male rats by adrenalectomy on the pancreatic zymogen granules was studied. Zymogen granules were purified from control, sham-operated and adrenalectomized animals studied 1, 3 and 7 days after surgery. The zymogen granules were characterized by flow cytometry, and in each granule the size (based on the forward or low angle light scatter (FSC) parameter), membrane complexity (based on side or 90° light scatter (SSC) parameter) and amylase content were evaluated. Amylase content/DNA ratio in pancreatic homogenates was also analyzed. The zymogen granules of the control rats were found to be distributed in two populations: a major one – R1 (95·45 ± 1·21%) – containing zymogen granules with a smaller mean size and complexity, and a minor population - R2 (4·45 ± 0·24%) – the granules of which had a mean size which was larger and more complex. At day +1 after adrenalectomy the zymogen granules were significantly (P<0·05) smaller than those of control animals. The R2 zymogen granules were similar to those from R1 as regards their size, but were more complex, suggesting that the immediate effect of glucocorticoid deprivation is to induce a depletion of the larger granules presumably belonging to the R2 population. The amount of amylase per granule did not vary at day +1 after adrenalectomy, although the amylase content/size ratio per granule was significantly (P<0·001) increased. This mechanism could be explained in terms of the existence of a bypass defined in the adrenalectomized animals between the granular content and cytosolic enzymes. Prolongation of the adrenalectomy period to 3 and 7 days resulted in a progressive increase in zymogen granule size and complexity, both parameters showing similar characteristics to those of the controls at day +7 after adrenalectomy. However, the percentage of zymogen granules within the R1 and R2 populations was clearly different from that of controls since the R2 population was much more numerous (11·25 ± 0·75% and 15·25 ± 1·15% (adrenalectomized rats at days +3 and +7 respectively) versus 4·45 ± 0·24% (controls)). An increase in the content of amylase per DNA was observed in adrenalectomized rats at day +1 although this transient effect cannot be related to glucocorticoid deprivation because it was also observed in sham-operated rats (day +1). However, a significant reduction, nearly 64%, in the amylase content/DNA ratio is produced by the absence of glucocorticoids 7 days after adrenalectomy and this is associated with a reduction in the content of amylase in each individual zymogen granule which reaches a minimum 3 days after adrenalectomy. It should be noted that, despite this, the enzyme concentration in each granule remains constant as there is a parallel decrease in the zymogen granule amylase content and size.
Journal of Endocrinology (1995) 147, 431–440
Chloride channels in the apical membrane of thyroid epithelial cells are regulated by cyclic AMP
Authors: Bourke J R , Sand O , Abel K C , Huxham G J , Manley S W 
Published: December 2008

Abstract
Porcine thyroid epithelial cells cultured as a monolayer with their apical membranes facing the medium are known to absorb Na+ and to secrete the anions Cl− and HCO3−. Chloride channels were found in the apical membrane, and displayed a reversal potential close to the resting membrane potential, linear current–voltage relationships, a conductance at physiological temperature of 6·5 pS, and a small but significant permeability to HCO3−. Stimulation of ion transport with prostaglandin E2 or 8-(4-chlorophenylthio) adenosine 3′:5′-cyclic monophosphate promoted activation of Cl− channels in cell-attached patches, and excised patches were reactivated by exposure of their cytoplasmic surface to protein kinase A and ATP. Physiological temperatures were necessary for activation of Cl− channels in cell-attached patches. The channels exhibited sub-states with a conductance exactly half that of the full unit conductance, suggesting a dual-barrelled channel structure. It is concluded that the apical membrane of thyroid epithelial cells contains cyclic AMP-activated Cl− channels controlling anion transport.
Journal of Endocrinology (1995) 147, 441–448
Bioactive oxytocin in human and baboon corpora lutea
Authors: Khan-Dawood F S , Yang J , Anwer K , Dawood M Y 
Published: December 2008

Abstract
Oxytocin has been identified in both non-human primate and human corpora lutea of the menstrual cycle by RIA, immunocytochemistry and HPLC. Evidence for the transcription of the oxytocin gene in this tissue using PCR is available. Oxytocin receptors have been characterized by biochemical procedures. However, there is some debate as to whether the oxytocin identified in these tissues is biologically active and has a role in luteal function. In this study we have demonstrated that oxytocin isolated by gel chromatography of tissue extracts from the baboon and the human corpus luteum is biologically active as determined in a rat uterine bioassay. Since both oxytocin and its receptors are present in these tissues, it is suggested that oxytocin in the human and non-human primate corpora lutea has a functional role.
Journal of Endocrinology (1995) 147, 525–532