Sumiko Morimoto

Progesterone receptor isoforms expression pattern in the rat brain during the estrotts cycle

Progesterone receptor (PR) isoforms expression was determined in the hypothalamus, the preoptic area, the hippocampus and the frontal cerebral cortex of the rat at 12:00 h on each day of the estrous cycle by using reverse transcription coupled to polymerase chain reaction. Rats under a 14:10 h light-dark cycle, with lights on at 06:00 h were used. We found that PR-B isoform was predominant in the hypothalamus, the preoptic area and the frontal cerebral cortex. Both PR isoforms were similarly expressed in the hippocampus. The highest PR-B expression was found on proestrus day in the hypothalamus; on metestrus in the preoptic area; and on diestrus in the frontal cortex. We observed no changes in PR isoforms expression in the hippocampus during the estrous cycle. These results indicate that PR isoforms expression is differentially regulated during the estrous cycle in distinct brain regions and that PR-B may be involved in progesterone actions upon the hypothalamus, the preoptic area and the frontal cortex of the rat.

Testosterone Effect on Insulin Content, Messenger Ribonucleic Acid Levels, Promoter Activity, and Secretion in the Rat1

Coexistence of hyperinsulinemia and hyperandrogenism in women has been frequently described. Most of the studies addressing this issue have focused on the mechanisms by which insulin produces hyperandrogenism. In the present study, we analyzed the effects of testosterone in vivo and in vitro upon insulin gene expression and release in the rat. Our studies demonstrate that testosterone increases insulin messenger RNA (mRNA) levels in vitro as well as in vivo. In both prepuberal and intact adult rats, serum testosterone concentrations were positively correlated with insulin mRNA levels and insulin concentration in serum. Testosterone deprivation after gonadectomy decreased both insulin gene expression and serum insulin concentration. Insulin mRNA levels were partially restored after 3 days of testosterone administration and serum insulin was 80% and 27% above baseline values at 5 and 7 days posttreatment. Primary cultured pancreatic islets treated with the sexual steroid increased about 80% insulin mRNA, as well as protein, and release. In transfected islets, testosterone increased the activity of the -410 bp rat insulin promoter I by 154%. These data demonstrate that testosterone has a direct effect upon pancreatic islet function by favoring insulin gene expression and release.

Progesterone receptor isoforms expression in the prepuberal and adult male rat brain

Progesterone receptor (PR) isoforms expression was determined in several regions of the prepuberal and adult male rat brain by using reverse transcription coupled to polymerase chain reaction. Rats under a 14:10-h light-dark cycle, with lights on at 0600 h were used. We found that in the hypothalamus of prepuberal animals the expression of both PR isoforms was similar, whereas PR-A expression was higher than that of PR-B in adults. In the cerebellum PR-B expression was predominant in both prepuberal and adult rats. In both ages PR-A and PR-B exhibited a non-significant tendency to be predominant in the hippocampus and the preoptic area respectively. In the frontal cortex and the olfactory bulb PR isoforms were expressed at a similar level. These results indicate a differential expression pattern of PR isoforms in the male rat brain and suggest that the tissue-specific expression of PR-A and PR-B is important for the appropriate response of each cerebral region to progesterone.

The Protective Effect of Testosterone on Streptozotocin-induced Apoptosis in β Cells Is Sex Specific

Objectives: To investigate the protective role of steroid hormones on streptozotocin (STZ)-induced apoptosis in rat pancreatic &bgr; cells. Methods: Two sets of experiments were performed. In the first, male rats were orchidectomized and substituted 72 hours later with testosterone, estradiol, or progesterone, and 24 hours later, administered with STZ. Subjects were killed 6 hours later, and apoptosis was determined in sections of the pancreas. In the second experiment, male or female rats were gonadectomized, were further substituted with testosterone, and then administered STZ. Six hours later, the animals were killed, and apoptosis, as well as immunoreactive expression of insulin, catalase, or Cu/Zn superoxide dismutase, was determined in sections of the pancreas. In addition, gonadectomized male or female subjects were substituted with testosterone and administered STZ, and 24 hours later, serum glucose and insulin were measured. Results: It was found that the cytoprotective effect was only shown in testosterone-treated male rats but not progesterone- or estradiol-treated male rats. In addition, the effect was seen in male rats but not in female rats, and there was an inverse correlation between apoptotic index and antioxidant enzyme immunoreactivity. Conclusions: The cytoprotective effect of testosterone is sex specific and is related to the induction of antioxidant enzyme activities in pancreatic &bgr; cells.

Administration of bisphenol A to dams during perinatal period modifies molecular and morphological reproductive parameters of the offspring.

Bisphenol A (BPA) is an estrogenic agonist compound that induces changes in diverse reproductive parameters in rats. The aim of the present study was to determine the effects of BPA given in drinking water containing 10mg/L (approximate dose 1.2mg/kg BW/day), administered chronically to rats during pregnancy and lactation, on reproductive tract parameters of the offspring. 79.2% of the female offspring from BPA-treated mothers presented irregular estrous cycles. As compared to the control group, a significant increase in the thickness of the uterine epithelia and stroma was observed in the BPA group. Additionally, 60% of the female offspring from BPA mothers did not undergo abundant uterine epithelial apoptosis during the estrus phase of the cycle while control animals did. In addition, a down regulation of ERα expression was observed in epithelial cells on estrus day. The results indicate that BPA, when administered chronically in water beverages to dams, modifies the reproductive cycle of the offspring during young adulthood.

Insulin gene expression pattern in rat pancreas during the estrous cycle

Sex steroid hormones influence insulin homeostasis and glucose metabolism, estradiol (E2) and progesterone (P4) induce changes in both fasting and postprandial insulinemia in rodents, however, insulin gene expression during estrous cycle is unknown. The aim of the present study was to determine an insulin gene expression pattern during the estrous cycle in the rat. Groups of 6 adult rats in each day of the estrous cycle were used. Serum P4, E2, testosterone (T) and insulin concentrations were determined by radioimmunoassay (RIA). A Northern blot analysis was performed to assess insulin gene expression in pancreatic tissue. We found a marked variation in insulin gene expression during the estrous cycle. The highest insulin expression was observed during the proestrus day. Interestingly, E2 and P4 but not T levels were correlated with changes in insulin mRNA content. The variations in serum insulin during the cycle were correlated with its mRNA content in pancreas. The overall results showed variations in serum insulin and insulin gene expression during estrous cycle of the rat that correlated with circulating E2 and P4 levels.

Androgen receptor in the rat pancreas: genetic expression and steroid regulation.

Androgens influence the incidence and prevalence of pancreatic cancer in humans and animal models. To our knowledge there has been molecular demonstration of the presence of neither the androgen receptor (AR) nor transcripts of the AR gene. Reverse-transcription polymerase chain reaction (RT-PCR)-Southern blotting was employed for molecular detection and measurement of the androgen receptor messenger ribonucleic acid (AR mRNA) in pancreas. Total RNA obtained from pancreas, prostate, seminal vesicles, and testes of neonatal and adult male and female rats, as well as castrated males substituted with testosterone cyclopentylate, was analyzed by Northern blot technique. Positive hybridization to AR cDNA was obtained in all tissues assayed but not in the pancreas. However, a clear AR 32P cDNA hybridization signal was obtained in pancreatic tissues after cDNA synthesis using RT-PCR-Southern blotting. The levels of the AR transcripts obtained by RT-PCR in the various pancreatic samples were as follows: adult females and neonatal animals > castrated adult males > adult males > castrated adult males substituted with testosterone. These results indicated that the pancreatic tissue possessed transcriptional activity of the AR gene, although to a lesser extent than the typical androgen responsive tissues (prostate and seminal vesicles). In conclusion, transcriptional activity of the AR gene in the pancreas seemed to be modulated by the androgenic milieu in the tissue similar to that reported for the classical androgen-responsive organs.

Role of progesterone receptors during postpartum estrus in rats.

We studied the role of progesterone receptor (PR) in the display of female sexual behavior during postpartum estrus in rats. Adult female rats were treated with the PR antagonist, RU486 (1.25 and 5 mg), 3 h after parturition and sexual behavior was evaluated throughout the first postpartum day. Estradiol and progesterone serum levels changed during the first 24 h postpartum. The highest estradiol and progesterone levels were found at 9 and 12 h postpartum, respectively. The predominant PR isoform in the hypothalamus and the preoptic area was PR-A during postpartum day. The content of PR-A increased at 6 h postpartum in the hypothalamus and the preoptic area, and decreased in both regions at 9 h. PR-B content only increased in the preoptic area at 12 h postpartum. The highest display of lordotic and proceptive behaviors were found at 12 h postpartum. The treatment with 1.25 and 5 mg of RU486 respectively reduced lordosis by 61% and 92% at 12 h postpartum. These results suggest that PR is essential in the display of postpartum estrus in rats.

Sex steroids effects on the endocrine pancreas

The endocrine pancreas is central in the physiopathology of diabetes mellitus. Nutrients and hormones control endocrine pancreatic function and the secretion of insulin and other pancreatic islet hormones. Although the pancreas is not usually considered as a target of steroids, increasing evidence indicates that sex steroid hormones modify pancreatic islet function. The biological effects of steroid hormones are transduced by both, classical and non-classical steroid receptors that in turn produce slow genomic and rapid non-genomic responses. In this review, we focused on the effects of sex steroid hormones on endocrine pancreatic function, with special emphasis in animal studies.

Molecular mechanism of cell proliferation in rodent uterus during the estrous cycle

The rodent uterus is a widely studied target tissue for sexual steroid hormone action. The aim of the present study was to assess the molecular mechanism that participates in the initiation of cell proliferation of the rat uterine epithelial cells during the estrus (E)-metestrus (M) transition. Cell proliferation, ERalpha, c-fos, cyclin D1 and D3, cdk4, and cdk6 proteins were assessed in these animals by immunohistochemistry. Estradiol (E(2)) and progesterone (P(4)) plasma levels were assessed by RIA. The results indicate that the glandular epithelium starts to proliferate at 21:00 h on estrus day, and initiates at least 3h before the luminal epithelium does. Fos expression was markedly increased during the afternoon of estrus day, and its increase was in parallel to ERalpha expression. Interestingly, both, cyclin D1 and D3 were abundantly expressed in the luminal and glandular epithelia, and nuclear immunolabelling of cyclin D1 and D3 precedes BrdU incorporation in the cell. cdk4 and cdk6 were localized in the nuclei in both epithelia throughout the studied time course. In addition, cdk4 was more abundant throughout estrus and metestrus days than cdk6. The overall results indicate that ERalpha, Fos and cyclins D1 and D3, cdk4 and cdk6 are expressed in both glandular and luminal epithelia of the rat uterus during the E-M transition. In conclusion, there is a good correlation between sequential expression of these proteins and cell cycle progression in the rat uterine epithelial cells during the estrous cycle. However, the differences observed in the cellular localization, time course of expression and the cellular types that express both cyclins between physiological and pharmacological conditions, demonstrated different mechanisms of regulation and should be due to the complex hormonal milieu during the estrous cycle.