We investigated the gestational expression of androgen receptor (AR) and defined its regulation and that of its co-repressors, PSF and p54nrb, by steroid hormones and myometrial stretch in vivo in pregnant and non-pregnant rats. Our data demonstrate that, 1) myometrial AR expression decreases prior to term; 2) AR expression is up-regulated by MPA treatment and down-regulated by mechanical stretch; (3) myometrial PSF protein expression is down-regulated by estrogen signaling and by mechanical stretch, and up-regulated by androgen signaling; (4) while myometrial PSF mRNA expression is also down-regulated by stretch, the regulation by estrogen and P4 on PSF mRNA appear to be opposite to the effects on PSF protein. We conclude that the decreased androgen signaling in late pregnancy (as a result of decreased AR and PSF expression mediated by hormonal and mechanical signals) may contribute to the mechanisms leading to labor initiation.

We investigated the gestational expression of androgen receptor (AR) and defined its regulation and that of its co-repressors, PSF and p54nrb, by steroid hormones and myometrial stretch in vivo in pregnant and non-pregnant rats. Our data demonstrate that, 1) myometrial AR expression decreases prior to term; 2) AR expression is up-regulated by MPA treatment and down-regulated by mechanical stretch; (3) myometrial PSF protein expression is down-regulated by estrogen signaling and by mechanical stretch, and up-regulated by androgen signaling; (4) while myometrial PSF mRNA expression is also down-regulated by stretch, the regulation by estrogen and P4 on PSF mRNA appear to be opposite to the effects on PSF protein. We conclude that the decreased androgen signaling in late pregnancy (as a result of decreased AR and PSF expression mediated by hormonal and mechanical signals) may contribute to the mechanisms leading to labor initiation.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / O objetivo deste estudo foi avaliar o efeito da administração de leptina no lobo ventral da próstata de ratos adultos. Vinte ratos Wistar machos e adultos foram divididos em 2 grupos: L - animais foram injetados com 50 μL diária de leptina (8 μg / 100 g PC, subcutânea) durante quatro dias e C - animais receberam o mesmo volume de solução salina. Perfil lipídico e níveis séricos de testosterona foram avaliados. O lobo ventral da próstata foi processado para análise histomorfométrica. Expressão dos genes da aromatase, receptor de andrógeno, receptores de estrógeno (α e β) e as isoformas dos receptores de leptina longa (Ob-Rb) e curta (Ob-Ra) foram avaliados por PCR em tempo real. Proliferação celular foi avaliada por imuno-histoquímica com PCNA. Os dados foram expressos como média  erro padrão e analisados pelo teste t de Student. Níveis séricos de colesterol aumentaram (C = 39,7 4,2; L = 55,2 4,2, mg / dL, P ≤ 0,02) e de testosterona (C = 1,6 0,43; L = 0,6 0,15, ng / dL, P ≤ 0,03) diminuíram no grupo L. A análise histomorfométrica mostrou uma redução na densidade de células (C = 8868 242; L = 8.211 210, mm2; P ≤ 0,04), na área total (C = 0,24 0,026; L = 0,10 0,009, mm2; P ≤ 0,001) e na área interna dos ácinos (C = 0,16 0,009; L = 0,08 0,006, mm2; P ≤ 0,0002). Por outro lado, houve um aumento na altura do epitélio (C = 17,3 0,3; L = 22,8 0,2 m, P ≤ 0,0001) e no número de ácinos (C = 7,0 0,2; L = 8,7 0,1, mm2; P ≤ 0,0002). As análises histomorfométrica juntamente com os resultados imuno-histoquímicos para PCNA sugerem que a leptina aumenta a proliferação celular. Em relação à expressão gênica, o tratamento de leptina aumentou a expressão de todos os genes, mas ER-α, em mais de 200 vezes em comparação com a expressão no grupo C. Em conclusão, neste trabalho mostramos que a leptina tem um efeito direto sobre a próstata de ratos adultos levando a um aumento na proliferação celular e na expressão gênica da aromatase, receptor de androgênio, nas isoformas dos receptores de leptina e receptores de estrogênios alfa e beta que são importantes para a fisiologia normal do tecido prostático / The aim of this study was to evaluate the effect of leptin administration on the ventral prostate lobe of adult rat. Twenty adult male rats were divided into 2 groups: L - animals were daily injected with 50 μL of leptin (8 g / 100 g BW, subcutaneous) for four days and C -animals received the same volume of saline solution. Lipid profile and testosterone serum levels were evaluated. The prostate ventral lobe was processed for histomorphometric analysis. Gene expression of aromatase, androgen receptor, leptin and estrogen receptors isoforms was evaluated by real-time PCR. Cell proliferation was evaluated by PCNA immunohistochemistry. Data were expressed as mean  standard error and analyzed by students t-test. Serum levels of cholesterol (C = 39.7 4.2; L = 55.2 4.2, mg / dL; P ≤ 0.02) increased and testosterone (C = 1.6 0.43; L = 0.6 0.15, ng / dL; P ≤ 0.03) decreased in L group. The histomorphometric analysis showed a reduction in cell density (C = 8868 242; L = 8211 210, mm2; P ≤ 0.04), in total (C = 0.24 0.026; L = 0.10 0.009, mm2; P ≤ 0.001) and in the internal acini areas (C = 0.16 0.009; L = 0.08 0.006, mm2; P ≤ 0.0002). On the other hand, there was an increase in the epithelial height (C = 17.3 0.3; L = 22.8 0.2, m; P ≤ 0.0001) and in the number of acini (C = 7.0 0.2; L = 8.7 0.1, mm2; P ≤ 0.0002). The histomorphometric analyses together with PCNA immunohistochemistry results suggest that leptin increases cell proliferation. In relation to the gene expression, leptin treatment increased the expression of all genes, but ER-α, in more than 200 times compared to the expression in C group. In conclusion, in this paper we showed that leptin has a direct effect on the prostate gland of adult rats leading to an increase in proliferation and in the gene expression of aromatase, androgen, leptin and estrogen receptors isoforms that are important for the physiology of the prostate gland.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / O objetivo deste estudo foi avaliar o efeito da administração de leptina no lobo ventral da próstata de ratos adultos. Vinte ratos Wistar machos e adultos foram divididos em 2 grupos: L - animais foram injetados com 50 μL diária de leptina (8 μg / 100 g PC, subcutânea) durante quatro dias e C - animais receberam o mesmo volume de solução salina. Perfil lipídico e níveis séricos de testosterona foram avaliados. O lobo ventral da próstata foi processado para análise histomorfométrica. Expressão dos genes da aromatase, receptor de andrógeno, receptores de estrógeno (α e β) e as isoformas dos receptores de leptina longa (Ob-Rb) e curta (Ob-Ra) foram avaliados por PCR em tempo real. Proliferação celular foi avaliada por imuno-histoquímica com PCNA. Os dados foram expressos como média  erro padrão e analisados pelo teste t de Student. Níveis séricos de colesterol aumentaram (C = 39,7 4,2; L = 55,2 4,2, mg / dL, P ≤ 0,02) e de testosterona (C = 1,6 0,43; L = 0,6 0,15, ng / dL, P ≤ 0,03) diminuíram no grupo L. A análise histomorfométrica mostrou uma redução na densidade de células (C = 8868 242; L = 8.211 210, mm2; P ≤ 0,04), na área total (C = 0,24 0,026; L = 0,10 0,009, mm2; P ≤ 0,001) e na área interna dos ácinos (C = 0,16 0,009; L = 0,08 0,006, mm2; P ≤ 0,0002). Por outro lado, houve um aumento na altura do epitélio (C = 17,3 0,3; L = 22,8 0,2 m, P ≤ 0,0001) e no número de ácinos (C = 7,0 0,2; L = 8,7 0,1, mm2; P ≤ 0,0002). As análises histomorfométrica juntamente com os resultados imuno-histoquímicos para PCNA sugerem que a leptina aumenta a proliferação celular. Em relação à expressão gênica, o tratamento de leptina aumentou a expressão de todos os genes, mas ER-α, em mais de 200 vezes em comparação com a expressão no grupo C. Em conclusão, neste trabalho mostramos que a leptina tem um efeito direto sobre a próstata de ratos adultos levando a um aumento na proliferação celular e na expressão gênica da aromatase, receptor de androgênio, nas isoformas dos receptores de leptina e receptores de estrogênios alfa e beta que são importantes para a fisiologia normal do tecido prostático / The aim of this study was to evaluate the effect of leptin administration on the ventral prostate lobe of adult rat. Twenty adult male rats were divided into 2 groups: L - animals were daily injected with 50 μL of leptin (8 g / 100 g BW, subcutaneous) for four days and C -animals received the same volume of saline solution. Lipid profile and testosterone serum levels were evaluated. The prostate ventral lobe was processed for histomorphometric analysis. Gene expression of aromatase, androgen receptor, leptin and estrogen receptors isoforms was evaluated by real-time PCR. Cell proliferation was evaluated by PCNA immunohistochemistry. Data were expressed as mean  standard error and analyzed by students t-test. Serum levels of cholesterol (C = 39.7 4.2; L = 55.2 4.2, mg / dL; P ≤ 0.02) increased and testosterone (C = 1.6 0.43; L = 0.6 0.15, ng / dL; P ≤ 0.03) decreased in L group. The histomorphometric analysis showed a reduction in cell density (C = 8868 242; L = 8211 210, mm2; P ≤ 0.04), in total (C = 0.24 0.026; L = 0.10 0.009, mm2; P ≤ 0.001) and in the internal acini areas (C = 0.16 0.009; L = 0.08 0.006, mm2; P ≤ 0.0002). On the other hand, there was an increase in the epithelial height (C = 17.3 0.3; L = 22.8 0.2, m; P ≤ 0.0001) and in the number of acini (C = 7.0 0.2; L = 8.7 0.1, mm2; P ≤ 0.0002). The histomorphometric analyses together with PCNA immunohistochemistry results suggest that leptin increases cell proliferation. In relation to the gene expression, leptin treatment increased the expression of all genes, but ER-α, in more than 200 times compared to the expression in C group. In conclusion, in this paper we showed that leptin has a direct effect on the prostate gland of adult rats leading to an increase in proliferation and in the gene expression of aromatase, androgen, leptin and estrogen receptors isoforms that are important for the physiology of the prostate gland.

Abstract Human prostatic acid phosphatase (hPAP) was the first laboratory parameter used for prostate cancer diagnosis, whereas the mechanisms behind the androgen regulation and tissue-specific expression of this prostate epithelium-specific differentiation antigen are not yet clear. In this study, a transient transfection model and transgenic animal model have been set up for functional analysis of the promoter and first intron region of the hPAP gene. The promoter constructs covering the region-734/+467 of the gene were functional in both prostatic and nonprostatic cells. Although hPAP constructs included two putative AREs with in vitro AR-binding ability at -178 and +336, androgen treatment had little effect on the promoter activity of the gene in transiently transfected cells. The hPAP fragment -734/+467 could trigger the expression of the CAT reporter gene and restrict the expression mainly in the prostates of transgenic mice. The DNA-binding site with the sequence GAAAATATGATA of a regulatory protein involved in prostate-specific and androgen receptor-dependent gene expression was identified from rPB promoter. The exact same 12 bp sequence was found in the first intron +1144/+1155 of the hPAP gene. Five homologous sequence, A, B, C, D and E, were located in the -734/+467 region of the hPAP gene, where site C and E could bind the regulatory protein in EMSA. Deletion of site C decreased the transcriptional activities significantly compared to those of corresponding wild-type constructs in LNCaP cells when androgens were present. Deletion of site E or both sites D and E increased the promoter activity in LNCaP when androgens were absent. In conclusion, androgens could not directly regulate hPAP expression via receptor-binding to the AREs in LNCaP cells. The promoter and first intron fragment -734/+467 of the hPAP gene could direct and restrict the gene expression mainly in prostate epithelium. A prostatic regulatory protein binds to multiple sites with the GAAAATATGATA or homologous sequences along the regulatory areas of the hPAP gene with different affinities, modulating the prostate-specific expression of the gene in a bidirectional manner, depending on the hormone status.

No description available.

Hormis leur implication dans les fonctions de reproduction, de développement et du maintien des caractères mâles, les androgènes (principalement la testostérone et la dihydrotestostérone, DHT) sont des hormones stéroïdiennes capables d’influencer plusieurs structures et fonctions du système nerveux. En effet, durant le développement, les androgènes ont un effet masculinisant sur le système nerveux central (SNC) le rendant sexuellement dimorphique. Chez les rongeurs mâles adultes, la substance blanche est plus volumineuse et les oligodendrocytes, cellules myélinisantes du SNC, sont plus nombreux. Cette différence est abolie après castration des mâles ; ce qui suggère l'implication de la testostérone dans le dimorphisme des oligodendrocytes et de la myéline.D’une part, mon travail de thèse visait à démontrer l’implication des androgènes et de leur récepteur (AR) dans l’établissement de ce dimorphisme. Nos résultats confirment l'implication de la testostérone et démontrent que son effet est médié par AR. En effet les corps calleux (CC) des souris mâles adultes ayant un AR non fonctionnel dans l'ensemble de l'organisme (souris Tfm) ou invalidé spécifiquement dans les cellules neurales (souris ARNesCre), présentent 20 à 30% moins d'oligodendrocytes et de surfaces myélinisées que ceux des contrôles. En outre, nos résultats montrent que ce dimorphisme apparait dès le dixième jour postnatal. De manière intéressante, le traitement pharmacologique des souriceaux mâles par un antagoniste du AR (flutamide) et des souriceaux femelles par l’agoniste d'AR (la DHT), pendant les dix premiers jours après la naissance inverse respectivement leurs profils oligodendrocytaires, suggérant un rôle organisationnel d'AR dans la substance blanche.D’autre part, mon sujet consistait à montrer l'importance de la testostérone et du AR dans la réparation de la myéline dans un modèle de démyélinisation de la moelle épinière des souris par injection stéréotaxique de lysolécithine. Nos résultats montrent que le traitement pendant 4 semaines des animaux par la testostérone permet le recrutement des oligodendrocytes et la réparation de la myéline dans les zones lésées. Il est à noter (1) qu’en absence de la testostérone ou d'AR, la réparation de la myéline est inefficace et se fait par des composants de la myéline périphérique et (2)que la présence des astrocytes semble nécessaire pour l’effet remyélinisant de la testostérone. Afin de mieux comprendre le ou les mécanisme(s) d'action(s) de la testostérone et du AR dans les processus de myélinisation et de remyélinisation, nous avons réalisé une étude transcriptomique comparative entre les animaux lésés et traités ou non avec la testostérone pour déterminer les gènes cibles et les voies de signalisations impliquées dans ces processus. Les résultats permettront probablement de définir une nouvelle cible thérapeutique pour les maladies démyélinisantes telle que la sclérose en plaques. / Androgens (mainly testosterone and dihydrotestosterone, DHT) are steroid hormones that are involved in reproduction functions, development and maintenance of male characteristics. They can also influence several structures and functions of the nervous system. Indeed, during development, androgens have a masculinizing effect on the central nervous system (CNS) making it sexually dimorphic. In addition, in adult male rodents,the white matter is larger and presents more oligodendrocytes, myelinating cells of the CNS. This difference is abolished after castration of males ; witch suggests the involvement of testosterone in the dimorphism of oligodendrocytes and myelin.One aim of my thesis was to study the involvement of androgens and their receptor (AR) in the establishment of this dimorphism. Our results confirm that testosterone is involved and demonstrate that its effect is mediated by AR. Indeed, the corpus callosum (CC) of adult male mice having a non-functional AR in the entire body (Tfm mice) or invalidated specifically in neural cells, (ARNesCre mice) have 20 to 30% fewer oligodendrocytes and myelinated area than those of controls. Moreover, our results show that this dimorphism appears early during postnatal life. Interestingly, pharmacological treatment of male pups with an AR antagonist (flutamide) and female ones with an AR agonist (DHT) during the first ten days after the birth reverses their oligodendrocytic profiles. These results suggest an organizational role of the AR in the white matter development.The aim of the second part of my study was to investigate the importance of testosterone and the AR in myelin repair in a rodent model of spinal cord demyelinationby stereotactic injection of lysolecithin. Our results show that a 4 weeks testosterone treatment allows the recruitment of oligodendrocyte and myelin repair. Interestingly, in the absence of testosterone or the AR, myelin repair was ineffeciant and was done by components of peripheral myelin. Moreover, the presence of astrocytes seems necessary for the remyelinating effect of testosterone since myelin repair was confined to astrocyte populated area.An important goal of my work is to better understand the mechanism of action of testosterone and the AR in the process of myelin formation and repair. For this, we performed a comparative transcriptomic study between animals injected or not with LPC than treated or not with testosterone to determine new target genes and signaling pathways involved in these processes. The results will probably define a new therapeutic target for demyelinating diseases such as multiple sclerosis.

No description available.

Introduction: Prostate cancer is the most common type of cancer in Sweden and is often treated using antiandrogenic drug therapy. Two substances belonging to this class of pharmaceuticals are bicalutamide and flutamide. After excretion from the human body, the drug molecules enter the wastewater treatment plant (WWTP). The WWTPs are not effective enough to completely remove pharmaceutical residues, why presence of both bicalutamide and flutamide can be detected in WWTP effluent water. Previous findings: Antiandrogens have been reported to affect reproduction in adult fish, but studies regarding possible effects on the embryonic development of fish are few. Aim: The present study sought to investigate if exposure to bicalutamide or flutamide cause toxicity in the early developmental stages of zebrafish embryos, and whether negative effects occur within concentrations relevant to measured environmental levels. Method: A modified OECD FET-test was used, where additional sublethal endpoints were included and the time period for assessment extended to 144 hours post fertilization (hpf). In addition, a locomotor activity assay was performed at 144 hpf in order to observe any sub-lethal swimming behavioral effects. Results: High doses (10 mg/L) of flutamide led to 100% lethality of the zebrafish embryos but the results suggest no acute toxic effects in the high dose treatment group of bicalutamide, or of either flutamide or bicalutamide within in the low (0.1 mg/L) or intermediate (1 mg/L) treatment groups. Neither did the locomotor activity assay result in statistically significant results, although the pattern of swimming activity in the low dose groups suggests that behavioral developmental effects could be present. Conclusions: High doses of flutamide caused mortality of the embryos, but no lethal or sublethal effects were present at environmentally relevant concentrations. The modest outcome of present study however suggests that further investigation of behavioral developmental effects of antiandrogens could be of future relevance. Analysis of the expression of genes related to neuronal growth, memory and other cognitive behaviors associated with behavioral changes, would then be of interest for further studies.

No description available.