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1.
Int J Mol Sci ; 21(16)2020 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-32796585

RESUMO

The orofacial pain pathway projects to the parabrachial and amygdala, and sex steroids have been shown to affect neuronal activity in these regions. GABA positive cells in the amygdala are influenced by sex steroid metabolites to affect pain, and sex steroids have been shown to alter the expression of genes in the parabrachial, changing neuronal excitability. Mechanisms by which sex steroids affect amygdala and parabrachial signaling are unclear. The expression of genes in the parabrachial and amygdala in diestrus (low estradiol) and proestrus (high estradiol) female rats were evaluated in this study. First, varicella zoster virus was injected into the whisker pad of female rats to induce a pain response. Second, gene expression was quantitated using RNA-seq one week after injection. Genes that had the greatest change in expression and known to function in pain signaling were selected for the quantitation of protein content. Protein expression of four genes in the parabrachial and seven genes in the amygdala were quantitated by ELISA. In the parabrachial, neurexin 3 (Nrnx3) was elevated at proestrus. Nrnx3 has a role in AMPA receptor and GABA signaling. Neuronatin (Nnat) and protein phosphatase, Mg2+/Mn2+ dependent 1E (Ppm1e) were elevated in the parabrachial of diestrus animals both genes having a role in pain signaling. Epoxide hydroxylase (Ephx2) was elevated in the parabrachial at proestrus and the vitamin D receptor (Vdr) was elevated in the amygdala. Ephx2 antagonists and vitamin D have been used to treat neuropathic pain. In conclusion, sex steroids regulate genes in the parabrachial and amygdala that might result in the greater pain response observed during diestrus.


Assuntos
Tonsila do Cerebelo/metabolismo , Diestro/genética , Regulação da Expressão Gênica , Herpesvirus Humano 3/fisiologia , Injeções , Proestro/genética , Animais , Epóxido Hidrolases/metabolismo , Feminino , Proteínas do Tecido Nervoso/metabolismo , Neuralgia/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Receptores de Calcitriol/metabolismo
2.
Physiol Genomics ; 50(9): 714-723, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29775429

RESUMO

DNA methylation is an important epigenetic modification involved in the estrous cycle and the regulation of reproduction. Here, we investigated the genome-wide profiles of DNA methylation in porcine ovaries in proestrus and estrus using methylated DNA immunoprecipitation sequencing. The results showed that DNA methylation was enriched in intergenic and intron regions. The methylation levels of coding regions were higher than those of the 5'- and 3'-flanking regions of genes. There were 4,813 differentially methylated regions (DMRs) of CpG islands in the estrus vs. proestrus ovarian genomes. Additionally, 3,651 differentially methylated genes (DMGs) were identified in pigs in estrus and proestrus. The DMGs were significantly enriched in biological processes and pathways related to reproduction and hormone regulation. We identified 90 DMGs associated with regulating reproduction in pigs. Our findings can serve as resources for DNA methylome research focused on porcine ovaries and further our understanding of epigenetically regulated reproduction in mammals.


Assuntos
Metilação de DNA/genética , Estro/genética , Genoma , Ovário/metabolismo , Proestro/genética , Suínos/genética , Animais , Cromossomos de Mamíferos/genética , Feminino , Ontologia Genética , Reprodutibilidade dos Testes , Reprodução/genética
3.
Reprod Fertil Dev ; 29(10): 1971-1981, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27997334

RESUMO

Kisspeptin is crucial for the generation of the circadian-gated preovulatory gonadotrophin-releasing hormone (GnRH)-LH surge in female rodents, with expression in the anteroventral periventricular nucleus (AVPV) peaking in the late afternoon of pro-oestrus. Given kisspeptin expression is established before puberty, the aim of the present study was to investigate kisspeptin and clock gene rhythms during the neonatal period. Anterior and posterior hypothalami were collected from C57BL/6J mice on Postnatal Days (P) 5, 15 and 25, at six time points across 24h, for analysis of gene expression by reverse transcription-quantitative polymerase chain reaction. Expression of aryl hydrocarbon receptor nuclear translocator-like gene (Bmal1) and nuclear receptor subfamily 1, group D, member 2 (Rev-erbα) in the anterior hypothalamus (containing the suprachiasmatic nucleus) was not rhythmic at P5 or P15, but Bmal1 expression exhibited rhythmicity in P25 females, whereas Rev-erbα expression was rhythmic in P25 males. KiSS-1 metastasis-suppressor (Kiss1) expression did not exhibit time-of-day variation in the anterior (containing the AVPV) or posterior (containing the arcuate nucleus) hypothalami in female and male mice at P5, P15 or P25. The data indicate that the kisspeptin circadian peak in expression observed in the AVPV of pro-oestrous females does not manifest at P5, P15 or P25, likely due to inadequate oestrogenic stimuli, as well as incomplete development of clock gene rhythmicity before puberty.


Assuntos
Proteínas CLOCK/metabolismo , Ritmo Circadiano/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Hipotálamo/metabolismo , Kisspeptinas/metabolismo , Fatores de Transcrição ARNTL/genética , Fatores de Transcrição ARNTL/metabolismo , Animais , Proteínas CLOCK/genética , Feminino , Kisspeptinas/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Membro 1 do Grupo D da Subfamília 1 de Receptores Nucleares/genética , Membro 1 do Grupo D da Subfamília 1 de Receptores Nucleares/metabolismo , Proestro/genética , Proestro/metabolismo , Receptores de Kisspeptina-1/genética , Receptores de Kisspeptina-1/metabolismo , Fatores Sexuais
4.
J Cell Physiol ; 231(5): 1057-64, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26388520

RESUMO

Pain can vary over the estrous cycle as a result of changes in estradiol concentration but the mechanism causing this variation is unclear. Because the thalamus is important in pain control, gene expression in the lateral thalamus (ventral posteromedial, ventral posterolateral, reticular thalamic nuclei) was screened at different phases of the estrous cycle. Gene expression changes in Sprague-Dawley rats were further analyzed by real-time PCR and ELISA and plasma estradiol levels were measured by RIAs at different phases of the estrous cycle. Our results indicated that both the RNA and protein expression of glutamate decarboxylase 1 and 2 (GAD1, GAD2), GABA(A) receptor-associated protein like 1 (GABARAPL1), and vesicular GABA transporter (VGAT) significantly increased in the lateral thalamus when plasma estradiol levels were elevated. Estradiol levels were elevated during the proestrus and estrus phases of the estrous cycle. Estrogen receptor α (ERα) was observed to be co-localized in thalamic cells and thalamic infusion of an ERα antagonist significantly reduced GAD1 and VGAT transcript. GAD1, GAD2, GABARAPL1, and VGAT have been shown to effect neuronal responses suggesting that attenuation of pain during the estrous cycle can be dependent, in part, through estradiol induced changes in thalamic gene expression.


Assuntos
Estro/genética , Proestro/genética , Transdução de Sinais/genética , Tálamo/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Estradiol/sangue , Receptor alfa de Estrogênio/antagonistas & inibidores , Receptor alfa de Estrogênio/metabolismo , Feminino , Imunofluorescência , Regulação da Expressão Gênica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley
5.
Genet Mol Res ; 15(1)2016 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-26909900

RESUMO

Frizzled 3 is an important receptor in the Wnt/ß-catenin pathway, a conserved signaling pathway that regulates gene expression and controls diverse developmental processes. However, the role of this protein during follicular development in the adult ovary is not known. The present study was designed to investigate the expression and localization of Frizzled 3 mRNA and protein during the estrous cycle in the mouse ovary through in situ hybridization (ISH), real-time quantitative polymerase chain reaction, immunohistochemistry and western blot. ISH results showed that in proestrus, high expression of Frizzled 3 was found in the granulosa and stroma with weak levels in the corpus luteum. In estrus and diestrus, the stroma had high Frizzled 3 expression, but levels were low in granulosa cells and corpus luteum. In the metestrus, moderate expression of Frizzled 3 was found in the stroma but low to no expression was found in luteal cells and follicles. The mRNA and protein levels of Frizzled 3 were found to be the highest in proestrus and diestrus compared to estrus and metestrus (P < 0.05), confirming the ISH results. During estrus and diestrus, high Frizzled 3 expression was observed in the stroma and moderate levels in granulosa cells, and during estrus and proestrus, low expression was seen in the oocyte cell membrane. The western blot results further confirmed this change during the estrous cycle. Together, these results indicate that Frizzled 3 is involved in regulating follicular development and oocyte maturation during the estrous cycle.


Assuntos
Corpo Lúteo/metabolismo , Receptores Frizzled/genética , Regulação da Expressão Gênica no Desenvolvimento , Oócitos/metabolismo , Folículo Ovariano/metabolismo , Animais , Corpo Lúteo/crescimento & desenvolvimento , Corpo Lúteo/ultraestrutura , Diestro/genética , Estro/genética , Feminino , Receptores Frizzled/metabolismo , Hibridização In Situ , Camundongos , Camundongos Endogâmicos ICR , Oócitos/crescimento & desenvolvimento , Oócitos/ultraestrutura , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/ultraestrutura , Proestro/genética , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Tempo
6.
Biol Reprod ; 93(1): 15, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25995272

RESUMO

Variations in mRNA levels and sources of metastin/kisspeptin, neurokinin B (NKB), dynorphin, and kisspeptin receptor GPR54 were examined in the ovaries of cycling rats. Kisspeptin and dynorphin mRNAs dramatically increased at 2000 h of the proestrous day. NKB mRNA also increased, but the peak was delayed by 6 h. GPR54 mRNA declined inversely with kisspeptin. Whole-ovary expressions of kisspeptin and dynorphin mRNAs, but not of NKB mRNA, were augmented by the administration of human chorionic gonadotropin (hCG). By means of laser-capture microdissection, kisspeptin mRNA was shown mostly in follicles at 2000 h of proestrus, whereas NKB and dynorphin were expressed mainly in interstitial tissues. GPR54 mRNA was detected equally in follicles, corpora lutea, and interstitial tissues. The hCG stimulated the follicular expression of kisspeptin and interstitial tissue expression of dynorphin mRNA. In primary cultures of granulosa cells prepared from equine chorionic gonadotropin-pretreated immature rats, hCG stimulated the expression of kisspeptin, dynorphin, and NKB mRNAs. Distortion of the corpus luteum and surrounding tissue borders was sometimes seen after intra-ovarian bursa administration of kisspeptin antagonist p234 for 3 days from proestrus. Progesterone production stimulated by hCG in granulosa cell culture was suppressed by p234. These data demonstrate that significant amounts of kisspeptin are synthesized in granulosa cells and dynorphin in interstitial tissues, in response to the proestrous luteinizing hormone surge, whereas granulosa cells also contain dynorphin and NKB, suggesting at least a role for kisspeptin in the luteinization of granulosa cells.


Assuntos
Regulação da Expressão Gênica , Células da Granulosa/metabolismo , Kisspeptinas/metabolismo , Luteinização/metabolismo , Hormônio Luteinizante/metabolismo , Proestro/metabolismo , Animais , Corpo Lúteo/efeitos dos fármacos , Corpo Lúteo/metabolismo , Dinorfinas/genética , Dinorfinas/metabolismo , Feminino , Células da Granulosa/efeitos dos fármacos , Kisspeptinas/genética , Luteinização/genética , Peptídeos/farmacologia , Proestro/efeitos dos fármacos , Proestro/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Kisspeptina-1
7.
Biol Reprod ; 89(1): 13, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23740946

RESUMO

Changes in the CD-1 mouse uterine transcriptome during proestrus and estrus were investigated to help elucidate mechanisms of uterine tissue remodeling during the estrus cycle and their regulation by estrogen and progesterone in preparation of the uterus for pregnancy. Mice were staged beginning at 6 weeks of age, and uterine horns were harvested after monitoring two estrus cycles. Microarray analysis of whole uterine horn RNA identified 2428 genes differentially expressed in estrus compared to proestrus, indicating there is extensive remodeling of mouse uterus during the estrus cycle, affecting ~10% of all protein-encoding genes. Many (~50%) of these genes showed the same differential expression in independent analyses of isolated uterine lumenal epithelial cells. Changes in gene expression associated with structural alterations of the uterus included remodeling of the extracellular matrix, changes in cell keratins and adhesion molecules, activation of mitosis and changes in major histocompatibility complex class II (MHCII) presentation, complement and coagulation cascades, and cytochrome P450 expression. Signaling pathways regulated during the estrus cycle, involving ligand-gated channels, Wnt and hedgehog signaling, and transcription factors with poorly understood roles in reproductive tissues, included several genes and gene networks that have been implicated in pathological states. Many of the molecular pathways and biological functions represented by the genes differentially expressed from proestrus to estrus are also altered during the human menstrual cycle, although not necessarily at the corresponding phases of the cycle. These findings establish a baseline for further studies in the mouse model to dissect mechanisms involved in uterine tissue response to endocrine disruptors and the development of reproductive tract diseases.


Assuntos
Estro/genética , Proestro/genética , Transcriptoma/genética , Útero/fisiologia , Animais , Feminino , Perfilação da Expressão Gênica , Camundongos , Modelos Animais , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase
8.
Horm Behav ; 57(2): 216-21, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19958772

RESUMO

The aim of this investigation was to evaluate overall DNA damage induced by experimental paradoxical sleep deprivation (PSD) in estrous-cycling and ovariectomized female rats to examine possible hormonal involvement during DNA damage. Intact rats in different phases of the estrous cycle (proestrus, estrus, and diestrus) or ovariectomized female Wistar rats were subjected to PSD by the single platform technique for 96 h or were maintained for the equivalent period as controls in home-cages. After this period, peripheral blood and tissues (brain, liver, and heart) were collected to evaluate genetic damage using the single cell gel (comet) assay. The results showed that PSD caused extensive genotoxic effects in brain cells, as evident by increased DNA migration rates in rats exposed to PSD for 96 h when compared to negative control. This was observed for all phases of the estrous cycle indistinctly. In ovariectomized rats, PSD also led to DNA damage in brain cells. No significant statistically differences were detected in peripheral blood, the liver or heart for all groups analyzed. In conclusion, our data are consistent with the notion that genetic damage in the form of DNA breakage in brain cells induced by sleep deprivation overrides the effects related to endogenous female sex hormones.


Assuntos
Dano ao DNA , Hormônios Esteroides Gonadais/metabolismo , Privação do Sono/genética , Privação do Sono/metabolismo , Sono REM , Animais , Sangue/metabolismo , Encéfalo/metabolismo , Corticosterona/metabolismo , Diestro/genética , Diestro/fisiologia , Estro/genética , Estro/fisiologia , Feminino , Fígado/metabolismo , Miocárdio/metabolismo , Ovariectomia , Proestro/genética , Proestro/fisiologia , Progesterona/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Testosterona/metabolismo
9.
Acta Histochem ; 122(8): 151630, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32992122

RESUMO

Visfatin is a crucial adipokine, which also regulates ovarian functions in many animals. Mice estrous cycle is characterized by a dynamic complex physiological process in the reproductive system. Expression of various factors changes during the estrous cycle in the ovary. To the best of our knowledge, no previous study has been conducted on the expression of visfatin in mice ovaries during the estrous cycle. Therefore, we investigated the localization and expression of visfatin protein in the ovary of mice during the estrous cycle. Western blot analysis showed the elevated expression of visfatin in proestrus and lowest in diestrus. Immunohistochemical localization of visfatin showed intense staining in the corpus luteum of proestrus and diestrus ovaries. Thecal cells, granulosa cells, and oocytes also showed the presence of visfatin. Expression of ovarian visfatin was correlated to BCL2 and active caspase3 expression and exhibited a significant positive correlation. Furthermore, in vivo inhibition of visfatin by FK866 in the proestrus ovary down-regulated active caspase3 and PCNA expression, and up-regulated the BCL2 expression. These results suggest the role of visfatin in the proliferation and apoptosis of the follicles and specific localization of visfatin in the corpus luteum also indicate its role in corpus luteum function, which may be in progesterone biosynthesis and regression of old corpus luteum. However, further study is required to support these findings. In conclusion, visfatin may also be regulating follicular growth during the estrous cycle by regulating proliferation and apoptosis.


Assuntos
Corpo Lúteo/metabolismo , Citocinas/genética , Células da Granulosa/metabolismo , Nicotinamida Fosforribosiltransferase/genética , Oócitos/metabolismo , Células Tecais/metabolismo , Acrilamidas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Caspase 3/genética , Caspase 3/metabolismo , Proliferação de Células/efeitos dos fármacos , Corpo Lúteo/citologia , Corpo Lúteo/efeitos dos fármacos , Corpo Lúteo/crescimento & desenvolvimento , Citocinas/antagonistas & inibidores , Citocinas/metabolismo , Diestro/efeitos dos fármacos , Diestro/genética , Inibidores Enzimáticos/farmacologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Células da Granulosa/citologia , Células da Granulosa/efeitos dos fármacos , Imuno-Histoquímica , Camundongos , Nicotinamida Fosforribosiltransferase/antagonistas & inibidores , Nicotinamida Fosforribosiltransferase/metabolismo , Oócitos/citologia , Oócitos/efeitos dos fármacos , Piperidinas/farmacologia , Proestro/efeitos dos fármacos , Proestro/genética , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Células Tecais/citologia , Células Tecais/efeitos dos fármacos
10.
J Psychopharmacol ; 23(4): 442-50, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18562442

RESUMO

Studies in people and animal models suggest that depression is influenced by natural fluctuations in the levels of 17beta-oestradiol (E(2)), as well as administration of E(2)-based therapies, such as selective oestrogen receptor modulators (SERMs). Elucidating the effects and mechanisms of E(2) is important to improve future E(2)-based therapeutics. An important question is whether effects of E(2) or SERMs for mood regulation act at the alpha or beta isoform of the oestrogen receptor (ER) because some of the unwanted trophic effects of E(2)-based therapies may involve actions at ERalpha, rather than ERbeta. In the present study, whether there are sex differences in depression-like behaviour of adult mice (experiment 1), and the effects of natural fluctuations in E(2) (experiment 2), or administration of E(2) or a SERM that has higher affinity for ERbeta than for ERalpha (diarylpropionitrile; DPN) to ovariectomised (experiment 3) wildtype and ERbeta knockout (betaERKO) mice were investigated. Results of this study supported our hypotheses that: there would be sex differences favouring males for depression-like behaviour and endogenous increases in, or exogenous administration of, E(2) or administration of an ERbeta SERM would decrease depression-like behaviour in wildtype, but not betaERKO, mice. In experiment 1, adult male mice spent less time immobile in the forced swim test (i.e., showed less depression-like behaviour) compared with female mice. In experiment 2, pro-oestrous (higher circulating E(2) levels), compared with dioestrous (lower circulating E(2) levels), mice had reduced immobility in the forced swim test; this effect was not observed in betaERKO mice. In experiment 3, administration of E(2) or DPN to ovariectomised wildtype, but not betaERKO, mice decreased immobility compared with vehicle administration, these data suggest that ERbeta may be required for some of the anti-depressant-like effects of E(2).


Assuntos
Depressão/tratamento farmacológico , Depressão/genética , Estradiol/uso terapêutico , Receptor beta de Estrogênio/genética , Nitrilas/uso terapêutico , Proestro/genética , Propionatos/uso terapêutico , Animais , Diestro/genética , Avaliação Pré-Clínica de Medicamentos , Receptor beta de Estrogênio/agonistas , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Caracteres Sexuais
11.
Biomed Res Int ; 2018: 9150723, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29662904

RESUMO

Estrus is an important factor for the fecundity of sows, and it is involved in ovulation and hormone secretion in ovaries. To better understand the molecular mechanisms of porcine estrus, the expression patterns of ovarian mRNA at proestrus and estrus stages were analyzed using RNA sequencing technology. A total of 2,167 differentially expressed genes (DEGs) were identified (P ≤ 0.05, |log2 Ratio| ≥ 1), of which 784 were upregulated and 1,383 were downregulated in the estrus compared with the proestrus group. Gene Ontology (GO) enrichment indicated that these DEGs were mainly involved in the cellular process, single-organism process, cell and cell part, and binding and metabolic process. In addition, a pathway analysis showed that these DEGs were significantly enriched in 33 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including cell adhesion molecules, ECM-receptor interaction, and cytokine-cytokine receptor interaction. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) confirmed the differential expression of 10 selected DEGs. Many of the novel candidate genes identified in this study will be valuable for understanding the molecular mechanisms of the sow estrous cycle.


Assuntos
Estro/genética , Perfilação da Expressão Gênica , Ovário/metabolismo , Proestro/genética , Análise de Sequência de RNA/métodos , Animais , Feminino , Ontologia Genética , Anotação de Sequência Molecular , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Transdução de Sinais/genética
12.
J Endocrinol ; 229(3): 307-18, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27068699

RESUMO

Kisspeptin, the neuropeptide product of the Kiss1 gene, is critical in driving the hypothalamic-pituitary-gonadal (HPG) axis. Kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) and arcuate nucleus (Arc) of the hypothalamus mediate differential effects, with the Arc regulating negative feedback of sex steroids and the AVPV regulating positive feedback, vital for the preovulatory surge and gated under circadian control. We aimed to characterize hypothalamic Kiss1 and Kiss1r mRNA expression in nonpregnant and pregnant mice, and investigate potential circadian regulation. Anterior and posterior hypothalami were collected from C57BL/6J mice at diestrus, proestrus, and days 6, 10, 14, and 18 of pregnancy, at six time points across 24h, for real-time PCR analysis of gene expression. Analysis confirmed that Kiss1 mRNA expression in the AVPV increased at ZT13 during proestrus, with a luteinizing hormone surge observed thereafter. No diurnal regulation was seen at diestrus or at any stage of pregnancy. Anterior hypothalamic Avp mRNA expression exhibited no diurnal variation, but Avpr1a peaked at 12:00h during proestrus, possibly reflecting the circadian input from the suprachiasmatic nucleus to AVPV Kiss1 neurons. Rfrp (Npvf) expression in the posterior hypothalamus did not demonstrate diurnal variation at any stage. Clock genes Bmal1 and Rev-erbα were strongly diurnal, but there was little change between diestrus/proestrus and pregnancy. Our data indicate the absence of the circadian input to Kiss1 in pregnancy, despite high gestational estradiol levels and normal clock gene expression, and may suggest a disruption of a kisspeptin-specific diurnal rhythm that operates in the nonpregnant state.


Assuntos
Ritmo Circadiano/fisiologia , Hipotálamo/fisiologia , Kisspeptinas/fisiologia , Prenhez/fisiologia , Animais , Arginina Vasopressina/genética , Ritmo Circadiano/genética , Feminino , Expressão Gênica , Hormônios/sangue , Kisspeptinas/genética , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , Prenhez/genética , Proestro/genética , Proestro/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Vasopressinas/genética
13.
Endocrinology ; 157(3): 1082-93, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26677881

RESUMO

Gonadotropes in the anterior pituitary gland are of particular importance within the hypothalamic-pituitary-gonadal axis because they provide a means of communication and thus a functional link between the brain and the gonads. Recent results indicate that female gonadotropes may be organized in the form of a network that shows plasticity and adapts to the altered endocrine conditions of different physiological states. However, little is known about functional changes on the molecular level within gonadotropes during these different conditions. In this study we capitalize on a binary genetic strategy in order to fluorescently label murine gonadotrope cells. Using this mouse model allows to produce an enriched gonadotrope population using fluorescence activated cell sorting to perform mRNA sequencing. By using this strategy, we analyze and compare the expression profile of murine gonadotropes in different genders and developmental and hormonal stages. We find that gonadotropes taken from juvenile males and females, from cycling females at diestrus and at proestrus, from lactating females, and from adult males each have unique gene expression patterns with approximately 100 to approximately 500 genes expressed only in one particular stage. We also demonstrate extensive gene-expression profile changes with up to approximately 2200 differentially expressed genes when comparing female and male development, juveniles and adults, and cycling females. Differentially expressed genes were significantly enriched in the GnRH signaling, calcium signaling, and MAPK signaling pathways by Kyoto Encyclopedia of Genes and Genomes analysis. Our data provide an unprecedented molecular view of the primary gonadotropes and reveal a high degree of molecular plasticity within the gonadotrope population.


Assuntos
Plasticidade Celular/genética , Ciclo Estral/genética , Gonadotrofos/metabolismo , Lactação/genética , RNA Mensageiro/genética , Maturidade Sexual/genética , Transcriptoma , Animais , Sinalização do Cálcio/genética , Diestro/genética , Diestro/metabolismo , Ciclo Estral/metabolismo , Feminino , Citometria de Fluxo , Hormônio Liberador de Gonadotropina/metabolismo , Lactação/metabolismo , Sistema de Sinalização das MAP Quinases/genética , Masculino , Camundongos , Proestro/genética , Proestro/metabolismo , Análise de Sequência de RNA , Transdução de Sinais/genética
14.
Endocrinology ; 156(8): 2999-3011, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25993523

RESUMO

The kisspeptin (Kp) neurons in the anteroventral periventricular nucleus (AVPV) are essential for the preovulatory LH surge, which is gated by circulating estradiol (E2) and the time of day. We investigated whether AVPV Kp neurons in intact female mice may be the site in which both E2 and daily signals are integrated and whether these neurons may host a circadian oscillator involved in the timed LH surge. In the afternoon of proestrous day, Kp immunoreactivity displayed a marked and transient decrease 2 hours before the LH surge. In contrast, Kp content was stable throughout the day of diestrus, when LH levels are constantly low. AVPV Kp neurons expressed the clock protein period 1 (PER1) with a daily rhythm that is phase delayed compared with the PER1 rhythm measured in the main clock of the suprachiasmatic nuclei (SCN). PER1 rhythm in the AVPV, but not in the SCN, exhibited a significant phase delay of 2.8 hours in diestrus as compared with proestrus. Isolated Kp-expressing AVPV explants from PER2::LUCIFERASE mice displayed sustained circadian oscillations of bioluminescence with a circadian period (23.2 h) significantly shorter than that of SCN explants (24.5 h). Furthermore, in AVPV explants incubated with E2 (10 nM to 1 µM), the circadian period was lengthened by 1 hour, whereas the SCN clock remained unaltered. In conclusion, these findings indicate that AVPV Kp neurons display an E2-dependent daily rhythm, which may possibly be driven by an intrinsic circadian clock acting in combination with the SCN timing signal.


Assuntos
Núcleo Hipotalâmico Anterior/metabolismo , Relógios Circadianos/genética , Kisspeptinas/genética , Animais , Diestro/efeitos dos fármacos , Diestro/genética , Diestro/metabolismo , Estradiol/metabolismo , Estradiol/farmacologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Hipotálamo/metabolismo , Kisspeptinas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Proestro/efeitos dos fármacos , Proestro/genética , Proestro/metabolismo , Núcleo Supraquiasmático/metabolismo
15.
Endocrinology ; 143(2): 577-86, 2002 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11796513

RESUMO

Activation of the ErbB-1 receptor is necessary for initiating mammalian female puberty by stimulating the release of LH-releasing hormone. It remains unclear whether ErbB-1 is also required in governing reproduction during adulthood and whether altered ErbB-1 signaling is linked to changes in gonadotropin secretion in aging females. The present study examined these issues. RT-PCR was employed to determine changes in ErbB-1 mRNA levels during proestrus in both young adult (YA) and middle-aged (MA) female rats. Before the LH surge, expression levels in the preoptic area of YA rats increased to a maximal value. No such increase in ErbB-1 mRNA was found in MA rats. This difference was confirmed by the analysis of in situ hybridization histochemistry, where a stronger mRNA signal was observed in the preoptic area of YA rats compared with MA females. ErbB-1 protein levels measured by Western blot reflected this difference. A peak level of ErbB-1 mRNA in the median eminence-arcuate nucleus was detected at 0800 h in YA rats, but it was delayed in MA animals. There were intense ErbB-1 mRNA-positive cells in the arcuate nucleus. Pharmacological blockade of ErbB-1 receptor-mediated signal transduction resulted in the disruption of estrous cyclicity in YA rats. These results indicate that ErbB-1 receptors are necessary for maintaining normal estrous cycles. Consequently, age-related alterations in hypothalamic ErbB-1 gene activity may contribute to a delayed preovulatory LH secretion in aging females. Thus, the ErbB-1 signaling system plays an important role in the control of female reproduction during adulthood.


Assuntos
Envelhecimento/genética , Receptores ErbB/genética , Ciclo Estral/fisiologia , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Envelhecimento/fisiologia , Animais , Western Blotting , Receptores ErbB/biossíntese , Ciclo Estral/genética , Feminino , Regulação da Expressão Gênica/genética , Regulação da Expressão Gênica/fisiologia , Hormônios Esteroides Gonadais/sangue , Hibridização In Situ , Proestro/genética , Proestro/fisiologia , Sondas RNA , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Receptores Proteína Tirosina Quinases/antagonistas & inibidores , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
16.
Eur J Endocrinol ; 132(6): 771-6, 1995 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-7788020

RESUMO

The present study examined the changes in mRNA levels of a pituitary-specific trans-acting factor, Pit-1, and prolactin during the rat estrous cycle. Total cytoplasmic RNA was analyzed by Northern blot and slot-blot hybridization to examine the prolactin mRNA level. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to examine the Pit-1 mRNA level. Proestrous and estrous prolactin mRNA levels were significantly higher than the metestrous and diestrous levels, whereas Pit-1 mRNA levels of the estrous and metestrous stages were about two- to threefold higher than those of the proestrous and diestrous stages. Proestrous Pit-1 mRNA levels increased gradually from 10.00 h to 20.00 h, while prolactin mRNA levels slightly decreased until 14.00 h but increased later until 20.00 h. During the rat estrous cycle, especially in the afternoon of the proestrous day, changes of prolactin mRNA levels seem to follow a prior increase of Pit-1 mRNA. Therefore, Pit-1 may be partly involved in the regulation of prolactin gene expression according to the rat estrous cycle. Estradiol administration to ovariectomized rats significantly increased both the mRNA levels of prolactin and Pit-1, which suggests that the gene expression of Pit-1 is regulated by estrogen through indirect extracellular pathways.


Assuntos
Proteínas de Ligação a DNA/genética , Estro/genética , Estro/metabolismo , Prolactina/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/genética , Animais , Sequência de Bases , Primers do DNA/genética , Estradiol/farmacologia , Estro/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Dados de Sequência Molecular , Ovariectomia , Proestro/genética , Proestro/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Transcrição Pit-1
17.
Brain Res ; 690(1): 117-20, 1995 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-7496797

RESUMO

Previous studies have revealed that in several animal models, N-methyl-D,L-Aspartate (NMA) stimulates LH secretion by acting at a suprapituitary site. In addition, NMDA receptor antagonists appear to block GnRH neuronal activation on the afternoon of proestrous as evidenced by the lack of c-Fos expression in the neurons and by the absence of an ovulatory LH surge. However, administration of NMA does not induce c-Fos or c-Jun expression in GnRH neurons. To better understand the effects of NMDA receptor activation on GnRH neuronal function, we examined whether GnRH neurons express the NMDA receptor in male rats, and in female rats during diestrus and proestrus, by performing double label in situ hybridization. An 35S-labeled cRNA probe for the NMDA receptor subunit (NMDAR1) was used to quantify NMDAR1 mRNA and a digoxigenin-labeled cRNA probe for GnRH was used to identify GnRH neurons. The data were quantified and expressed as grains/average cell area. In male and female rats, less than 5% of GnRH neurons expressed grain levels twice the minimum detectable level and were considered double-labeled. However, many non-GnRH neurons in the same areas as GnRH neurons expressed high levels of NMDAR1 mRNA. These results suggest that the effects of NMA on GnRH secretion are unlikely to be mediated solely by the activation of NMDA receptors on GnRH neurons. Given the widespread expression of NMDAR1 mRNA in the hypothalamus, it is possible that the stimulatory effects of NMA on GnRH neurons are indirect through activation of other neurons.


Assuntos
Hormônio Liberador de Gonadotropina/análise , Neurônios/química , Receptores de N-Metil-D-Aspartato/genética , Animais , Diestro/genética , Feminino , Expressão Gênica , Hibridização In Situ , Hormônio Luteinizante/metabolismo , Masculino , Proestro/genética , RNA Mensageiro/biossíntese , Ratos , Taxa Secretória/fisiologia
18.
J Anim Sci ; 72(8): 1964-70, 1994 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7982823

RESUMO

Puberty traits of 740 Yorkshire gilts from a selection experiment were analyzed. Heritabilities were estimated to be .32 for pubertal age, .23 for length of proestrus, .16 for length of standing estrus, .29 for ability to show standing reflex, and .24 for intensity of vulvar symptoms. Genetic correlations between length of proestrus, intensity of vulvar symptoms, and ability to show standing reflex were positive. There seemed to be a negative genetic correlation between percentage of lean and intensity of vulvar symptoms. There were negative genetic correlations between growth rate and length of standing estrus, and between growth rate and ability to show standing reflex.


Assuntos
Composição Corporal/genética , Estro/genética , Maturidade Sexual/genética , Suínos/genética , Animais , Feminino , Fenótipo , Proestro/genética , Suínos/crescimento & desenvolvimento , Suínos/fisiologia , Vulva/fisiopatologia
19.
J Am Vet Med Assoc ; 200(8): 1104-6, 1992 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-1607315

RESUMO

A stunted Miniature American Eskimo bitch that had signs of proestrus, which persisted for almost 8 months, had a 77,XO karyotype. Despite signs of proestrus, the ovaries were small and fibrous, and there was no evidence of ovarian follicle development or corpora lutea. Except for its juvenile appearance, the rest of the reproductive tract was grossly normal. Clinical signs in this bitch were similar to those in human beings with Turner's syndrome.


Assuntos
Doenças do Cão/genética , Monossomia , Proestro/genética , Cromossomo X , Animais , Cães , Feminino , Hormônio Foliculoestimulante/sangue , Genitália Feminina/patologia , Cariotipagem/veterinária , Hormônio Luteinizante/sangue
20.
Tsitol Genet ; 24(3): 37-40, 1990.
Artigo em Russo | MEDLINE | ID: mdl-2238098

RESUMO

Female rats at the stage of proestrus have been subjected to a stress effect--immobilization for an hour. Oocytes were cytogenetically analyzed and embryonal mortality was determined. It is shown that this stress effect at the end of the fist meiotic division of the oocytes increased the rate of aneuploid gametes and the level of embryonal mortality.


Assuntos
Aberrações Cromossômicas/genética , Oócitos/ultraestrutura , Proestro/genética , Estresse Psicológico/genética , Animais , Aberrações Cromossômicas/etiologia , Transtornos Cromossômicos , Perda do Embrião/etiologia , Perda do Embrião/genética , Feminino , Meiose/genética , Ratos , Ratos Endogâmicos , Restrição Física , Estresse Psicológico/complicações
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