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1.
Biol Reprod ; 101(1): 76-90, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31066903

RESUMO

Mechanisms of bovine corpus luteum (CL) maintenance during the second month of pregnancy have not been adequately investigated, despite significant reproductive losses. In the first month, interferon-tau is believed to suppress oxytocin-stimulated prostaglandin F2α (PGF) production, yet there are conflicting reports of circulating PGF metabolite (PGFM). In this study, characterization of PGFM and P4 occurred through continuous bihourly blood sampling in cows undergoing CL regression (day 18-21, n = 5), and during the first (day 18-21, n = 5) and second month (day 47-61; n = 16) of pregnancy. Cattle in the second month were assigned to control (n = 8) or oxytocin treatment (n = 8; three pulses to mimic luteolysis) to evaluate if oxytocin receptors were active. All cows but one (which had elevated PGFM prior to oxytocin treatment) maintained the pregnancy. Basal PGFM concentrations were low (11.6 ± 0.7 pg/mL) in the first month but increased 2.54-fold in the second month. Few (0.26 ± 0.12 pulses/day) PGFM pulses with low peak concentrations (28.8 ± 3.1 pg/mL) were observed during the first month of pregnancy, similar to cows not undergoing regression. However, in the second month, frequency (1.10 ± 0.26 pulses/day) and peak concentration (67.2 ± 5.0 pg/mL) of PGFM pulses increased, displaying similar frequency but lower peak PGFM than seen in regression (1.44 ± 0.14 pulses/day; 134.5 ± 18.9 pg/mL). Oxytocin treatment increased likelihood of PGFM pulses post-treatment and increased peak concentration (89.7 ± 10.1 pg/mL) in cows during the second month. Thus, cows have more PGFM pulses during second than first month of pregnancy, possibly induced by endogenous oxytocin, indicating suppression of PGF production is an important mechanism for CL maintenance during first but not second month of pregnancy.


Assuntos
Bovinos , Manutenção do Corpo Lúteo/fisiologia , Dinoprosta/metabolismo , Lactação/metabolismo , Luteólise/metabolismo , Metaboloma , Prenhez , Animais , Bovinos/metabolismo , Corpo Lúteo/metabolismo , Manutenção do Corpo Lúteo/metabolismo , Indústria de Laticínios , Feminino , Gravidez , Prenhez/metabolismo
2.
J Endocrinol ; 231(3): 223-233, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27679426

RESUMO

This study aimed to determine in the canine corpus luteum throughout the dioestrus (1) the influence of insulin on glucose uptake; (2) the regulation of genes potentially involved; and (3) the influence of hypoxia on glucose transporter expression and steroidogenesis, after treatment with cobalt chloride (CoCl2). Glucose uptake by luteal cells increased 2.7 folds (P < 0.05) in response to insulin; a phenomenon related to increased expression of glucose transporter (GLUT) 4 and phosphorylation of protein kinase B (AKT). The gene expression of insulin receptor and SLC2A4 (codifier of GLUT4) genes after insulin stimulation increased on day 20 post ovulation (p.o.) and declined on day 40 p.o. (P < 0.05). Regarding potentially involved molecular mechanisms, the nuclear factor kappa B gene RELA was upregulated on days 30/40 p.o., when SLC2A4 mRNA was low, and the interleukin 6 (IL6) gene was upregulated in the first half of dioestrus, when SLC2A4 mRNA was high. CoCl2 in luteal cell cultures increased the hypoxia-inducible factor HIF1A/HIF1A and the SLC2A4/GLUT4 expression, and decreased progesterone (P4) production and hydroxyl-delta-5-steroid dehydrogenase 3 beta (HSD3B) mRNA expression (P < 0.05). This study shows that the canine luteal cells are responsive to insulin, which stimulates glucose uptake in AKT/GLUT4-mediated pathway; that may be related to local activity of RELA and IL6. Besides, the study reveals that luteal cells under hypoxia activate HIF1A-modulating luteal function and insulin-stimulated glucose uptake. These data indicate that insulin regulates luteal cells' glucose disposal, participating in the maintenance and functionality of the corpus luteum.


Assuntos
Corpo Lúteo/metabolismo , Cães/metabolismo , Insulina/metabolismo , Animais , Transporte Biológico Ativo/efeitos dos fármacos , Cobalto/farmacologia , Manutenção do Corpo Lúteo/genética , Manutenção do Corpo Lúteo/metabolismo , Cães/genética , Feminino , Expressão Gênica/efeitos dos fármacos , Glucose/metabolismo , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 4/genética , Transportador de Glucose Tipo 4/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Insulina/fisiologia , Interleucina-6/genética , Interleucina-6/metabolismo , Células Lúteas/efeitos dos fármacos , Células Lúteas/metabolismo , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptor de Insulina/genética , Receptor de Insulina/metabolismo , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo
3.
Reprod Fertil Dev ; 28(3): 269-77, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25033944

RESUMO

The aim was to examine the effect of a single intramuscular (i.m.) injection of equine chorionic gonadotrophin (eCG) on Day 3 after oestrus on corpus luteum (CL) development, circulating progesterone and conceptus development in cross-bred beef heifers. In Experiment 1, heifers received: (1) saline, or a single i.m. injection of eCG on Day 3 at (2) 250IU (3) 500IU (4) 750IU or (5) 1000IU. Administration of eCG resulted in increased luteal tissue area and progesterone and oestradiol concentrations compared with controls. In Experiment 2, heifers received (1) a progesterone-releasing intravaginal device (PRID Delta) from Day 3 to 5 or (2) a PRID Delta from Day 3 to 5 plus a single injection of 750IU eCG on Day 3. In vitro-produced blastocysts (n=10 per recipient) were transferred on Day 7 and heifers were slaughtered on Day 14 to assess conceptus development. Administration of eCG reduced the number of short cycles (6.3% vs 31.3%) and increased mean luteal tissue weight (P=0.02). Insertion of a PRID Delta on Day 3 resulted in an elevation (P<0.05) in serum progesterone until removal on Day 5. Administration of eCG at the time of PRID Delta insertion resulted in higher progesterone levels (P<0.05) from Day 10 onwards. Conceptus dimensions were not affected. In conclusion, a single injection of eCG on Day 3 increased CL size and progesterone concentrations and, when given in conjunction with a progesterone-releasing device, appeared to reduce the number of short cycles, presumably due to its luteotrophic nature. The implications of the elevated oestradiol concentrations for embryo quality require further study.


Assuntos
Gonadotropina Coriônica/administração & dosagem , Manutenção do Corpo Lúteo/efeitos dos fármacos , Corpo Lúteo/efeitos dos fármacos , Fármacos para a Fertilidade Feminina/administração & dosagem , Gonadotropinas Equinas/administração & dosagem , Progesterona/administração & dosagem , Técnicas de Reprodução Assistida/veterinária , Animais , Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Bovinos , Corpo Lúteo/metabolismo , Manutenção do Corpo Lúteo/metabolismo , Relação Dose-Resposta a Droga , Esquema de Medicação , Quimioterapia Combinada , Implantação do Embrião/efeitos dos fármacos , Transferência Embrionária/veterinária , Estradiol/sangue , Feminino , Fertilização in vitro/veterinária , Injeções Intramusculares , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/metabolismo , Gravidez , Progesterona/sangue , Fatores de Tempo
4.
Biol Reprod ; 89(4): 91, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23966322

RESUMO

Normal pregnancy is supported by increased levels of progesterone (P4), which is secreted from ovarian luteal cells via enzymatic steps catalyzed by P450scc (CYP11A1) and HSD3B. The development and maintenance of corpora lutea during pregnancy, however, are less well understood. Here we used Cyp11a1 transgenic mice to delineate the steps of luteal cell differentiation during pregnancy. Cyp11a1 in a bacterial artificial chromosome was injected into mouse embryos to generate transgenic mice with transgene expression that recapitulated endogenous Cyp11a1 expression. Cyp11a1 transgenic females displayed reduced pregnancy rate, impaired implantation and placentation, and decreased litter size in utero, although they produced comparable numbers of blastocysts. The differentiation of transgenic luteal cells was delayed during early pregnancy as shown by the delayed activation of genes involved in steroidogenesis and cholesterol availability. Luteal cell mitochondria were elongated, and their numbers were reduced, with morphology and numbers similar to those observed in granulosa cells. Transgenic luteal cells accumulated lipid droplets and secreted less progesterone during early pregnancy. The progesterone level returned to normal on gestation day 9 but was not properly withdrawn at term, leading to delayed stillbirth. P4 supplementation rescued the implantation rates but not the ovarian defects. Thus, overexpression of Cyp11a1 disrupts normal development of the corpus luteum, leading to progesterone insufficiency during early pregnancy. Misregulation of the progesterone production in Cyp11a1 transgenic mice during pregnancy resulted in aberrant implantation, anomalous placentation, and delayed parturition.


Assuntos
Enzima de Clivagem da Cadeia Lateral do Colesterol/biossíntese , Decídua/enzimologia , Infertilidade Feminina/enzimologia , Células Lúteas/metabolismo , Luteinização/metabolismo , Progesterona/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Enzima de Clivagem da Cadeia Lateral do Colesterol/genética , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Manutenção do Corpo Lúteo/sangue , Manutenção do Corpo Lúteo/efeitos dos fármacos , Manutenção do Corpo Lúteo/metabolismo , Cruzamentos Genéticos , Decídua/efeitos dos fármacos , Decídua/metabolismo , Decídua/patologia , Implantação do Embrião/efeitos dos fármacos , Feminino , Terapia de Reposição Hormonal , Infertilidade Feminina/tratamento farmacológico , Infertilidade Feminina/metabolismo , Infertilidade Feminina/patologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Tamanho da Ninhada de Vivíparos/efeitos dos fármacos , Células Lúteas/efeitos dos fármacos , Células Lúteas/patologia , Luteinização/sangue , Luteinização/efeitos dos fármacos , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/ultraestrutura , Placentação/efeitos dos fármacos , Gravidez , Progesterona/deficiência , Progesterona/uso terapêutico , Organismos Livres de Patógenos Específicos
5.
Reprod Fertil Dev ; 25(1): 62-70, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23244829

RESUMO

Ongoing detailed investigations into embryo-maternal communication before implantation reveal that during early embryonic development a plethora of events are taking place. During the sexual cycle, remodelling and differentiation processes in the endometrium are controlled by ovarian hormones, mainly progesterone, to provide a suitable environment for establishment of pregnancy. In addition, embryonic signalling molecules initiate further sequences of events; of these molecules, prostaglandins are discussed herein as specifically important. Inadequate receptivity may impede preimplantation development and implantation, leading to embryonic losses. Because there are multiple factors affecting fertility, receptivity is difficult to comprehend. This review addresses different models and methods that are currently used and discusses their respective potentials and limitations in distinguishing key messages out of molecular twitter. Transcriptome, proteome and metabolome analyses generate comprehensive information and provide starting points for hypotheses, which need to be substantiated using further confirmatory methods. Appropriate in vivo and in vitro models are needed to disentangle the effects of participating factors in the embryo-maternal dialogue and to help distinguish associations from causalities. One interesting model is the study of somatic cell nuclear transfer embryos in normal recipient heifers. A multidisciplinary approach is needed to properly assess the importance of the uterine milieu for embryonic development and to use the large number of new findings to solve long-standing issues regarding fertility.


Assuntos
Blastocisto/fisiologia , Bovinos/fisiologia , Implantação do Embrião , Endométrio/fisiologia , Modelos Biológicos , Animais , Bovinos/embriologia , Biologia Computacional/métodos , Manutenção do Corpo Lúteo/metabolismo , Feminino , Técnicas de Transferência Nuclear/veterinária , Gravidez
6.
Reprod Biomed Online ; 25(6): 551-60, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23063822

RESUMO

Progesterone produced by the corpus luteum (CL) regulates the synthesis of various endometrial proteins required for embryonic implantation and development. Compromised CL progesterone production is a potential risk factor for prenatal development. Reactive oxygen species (ROS) play diverse roles in mammalian reproductive biology. ROS-induced oxidative damage and subsequent adverse developmental outcomes constitute important issues in reproductive medicine. The CL is considered to be highly exposed to locally produced ROS due to its high blood vasculature and steroidogenic activity. ROS-induced apoptotic cell death is involved in the mechanisms of CL regression that occurs at the end of the non-fertile cycle. Luteal ROS production and propagation depend upon several regulating factors, including luteal antioxidants, steroid hormones and cytokines, and their crosstalk. However, it is unknown which of these factors have the greatest contribution to the maintenance of CL integrity and function during the oestrous/menstrual cycle. There is evidence to suggest that antioxidants play important roles in CL rescue from luteolysis when pregnancy ensues. As luteal phase defect impacts fertility by preventing implantation and early conceptus development in livestock and humans, this review attempts to address the importance of ROS-scavenging antioxidant enzymes in the control of mammalian CL function and integrity. The corpus luteum (CL) is a transient endocrine organ that develops after ovulation from the ovulated follicle during each reproductive cycle. The main function of the CL is the production and secretion of progesterone which is necessary for embryonic implantation and development. Compromised CL progesterone production is a potential risk factor for prenatal development and pregnancy outcomes. Reactive oxygen species (ROS), which are natural by-products of cellular respiration and metabolism, play diverse roles in mammalian reproductive biology. ROS-induced oxidative damage and subsequent development of adverse pregnancy outcomes constitute important issues in reproductive medicine. Before the end of the first trimester, a high rate of human and animal conceptions end in spontaneous abortion and most of these losses occur at the time of implantation in association with ROS-induced oxidative damage. Every cell in the body is normally able to defend itself against the oxidative damage caused by the ROS. The cellular antioxidant enzymes constitute the first line of defence against the toxic effects of ROS. The CL is considered to be highly exposed to locally produced ROS due to its high blood vasculature and metabolic activity. There is now evidence to suggest that cellular antioxidants play important roles in CL rescue from regression when pregnancy ensues. As defective CL function impacts fertility by preventing implantation and early conceptus development in livestock and humans, this review attempts to address the importance of antioxidant enzymes in the control of mammalian CL function and integrity.


Assuntos
Manutenção do Corpo Lúteo/metabolismo , Corpo Lúteo/enzimologia , Estresse Oxidativo , Oxirredutases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Animais , Apoptose , Hipóxia Celular , Corpo Lúteo/irrigação sanguínea , Corpo Lúteo/metabolismo , Feminino , Humanos , Gravidez , Progesterona/metabolismo
7.
Reprod Biol Endocrinol ; 10: 72, 2012 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-22950660

RESUMO

BACKGROUND: Studies suggested that microRNAs influence cellular activities in the uterus including cell differentiation and embryo implantation. In assisted reproduction cycles, luteal phase support, given to improve endometrial characteristics and to facilitate the implantation process, has been a standard practice. The effect of different types of luteal phase support using steroid hormones in relation to endometrial miRNA profiles during the peri-implantation period has not seen described. This study was designed to evaluate the expression of miRNAs during the luteal phase following controlled ovarian stimulation for IVF and the influence of different luteal phase support protocols on miRNA profiles. METHODS: The study was approved by the Johns Hopkins Hospital Institutional Review Board. Endometrial biopsies were obtained on the day of oocyte retrieval from 9 oocyte donors (group I). An additional endometrial biopsy was obtained 3-5 days later (Group II) after the donors were randomized into three groups. Group IIa had no luteal-phase support, group IIb had luteal support with micronized progesterone (P), and Group IIc had luteal support with progesterone plus 17-beta-estradiol (P + E). Total RNA was isolated and microarray analysis was performed using an Illumina miRNA expression panel. RESULTS: A total of 526 miRNAs were identified. Out of those, 216 miRNAs were differentially regulated (p < 0.05) between the comparison groups. As compared to the day of retrieval, 19, 11 and 6 miRNAs were differentially regulated more than 2 fold in the groups of no support, in the P support only, and in the P + E support respectively, 3-5 days after retrieval. During the peri-implantation period (3-5 days after retrieval) the expression of 33 and 6 miRNAs increased, while the expression of 3 and 0 miRNAs decreased, in the P alone and in the P + E group respectively as compared to the no steroid supplementation group. CONCLUSION: Luteal support following COS has a profound influence on miRNA profiles. Up or down regulation of miRNAs after P or P + E support suggest a role(s) of luteal support in the peri-implantation uterus in IVF cycles through the regulation of associated target genes.


Assuntos
Manutenção do Corpo Lúteo/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Endométrio/efeitos dos fármacos , Fase Luteal/efeitos dos fármacos , MicroRNAs/metabolismo , Indução da Ovulação , Regulação para Cima/efeitos dos fármacos , Adulto , Manutenção do Corpo Lúteo/metabolismo , Endométrio/metabolismo , Estradiol/farmacologia , Estrogênios/farmacologia , Feminino , Perfilação da Expressão Gênica , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Antagonistas de Hormônios/farmacologia , Humanos , Fase Luteal/metabolismo , MicroRNAs/genética , Análise de Sequência com Séries de Oligonucleotídeos , Doação de Oócitos , Gravidez , Progesterona/farmacologia , Progestinas/farmacologia , Doadores de Tecidos , Adulto Jovem
8.
Fertil Steril ; 98(5): 1112-7, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22819186

RESUMO

Luteal phase deficiency (LPD) has been described in healthy normally menstruating women and in association with other medical conditions. While progesterone is important for the process of implantation and early embryonic development, LPD, as an independent entity causing infertility, has not been proven.


Assuntos
Manutenção do Corpo Lúteo , Infertilidade Feminina/etiologia , Fase Luteal , Ovário/fisiopatologia , Fatores Etários , Envelhecimento/metabolismo , Biomarcadores/sangue , Biópsia , Manutenção do Corpo Lúteo/metabolismo , Implantação do Embrião , Feminino , Humanos , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/metabolismo , Infertilidade Feminina/fisiopatologia , Infertilidade Feminina/terapia , Fase Luteal/metabolismo , Obesidade/complicações , Ovário/metabolismo , Valor Preditivo dos Testes , Gravidez , Progesterona/sangue , Medição de Risco , Fatores de Risco
9.
J Reprod Dev ; 58(4): 445-52, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22522231

RESUMO

Persistence of the corpus luteum (CL) in cattle usually occurs during the puerperium and is associated with interference of prostaglandin (PG) F(2α) release from the uterus. The objective of the present study was to determine for the first time the gene expressions in the persistent CL compared with the CL of pregnancy and cyclic CL. Three types of CL biopsy samples were collected from 32 lactating Holstein cows: (1) CL persisting for 29 to 33 days after the first ovulation postpartum (persistent CL, n=9), (2) CL between days 29 and 33 of early pregnancy (CL of pregnancy, n=8) and (3) CL between days 10 and 13 of the estrous cycle (cyclic CL, n=27). mRNA expression of 2',5'-oligoadenylate synthetase-1 was upregulated only in the CL of pregnancy, confirming exposure to interferon-τ (IFNT) produced by trophoblasts in pregnant cows. mRNA expressions of immune tolerance-related factors (PGES and forkhead/winged helix transcription factor 3) were upregulated in the CL of pregnancy but not in the persistent CL, suggesting that IFNT controls upregulation of these genes. mRNA expression relating to some of the major systems such as lymphangiogenesis, inflammation and apoptosis were similarly upregulated in the persistent CL and the CL of pregnancy but not in the cyclic CL. The results suggest that the persistent CL may survive for a long period without changes in local immune tolerance but develops several major systems required for CL maintenance similar to the CL of pregnancy.


Assuntos
Bovinos/fisiologia , Manutenção do Corpo Lúteo/metabolismo , Corpo Lúteo/metabolismo , Ciclo Estral/metabolismo , Regulação da Expressão Gênica , Luteólise , Manutenção da Gravidez , 2',5'-Oligoadenilato Sintetase/genética , 2',5'-Oligoadenilato Sintetase/metabolismo , Animais , Animais Endogâmicos , Indústria de Laticínios , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica , Tolerância Imunológica , Oxirredutases Intramoleculares/genética , Oxirredutases Intramoleculares/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Período Pós-Parto , Gravidez , Prostaglandina-E Sintases , RNA Mensageiro/metabolismo
10.
Am J Physiol Endocrinol Metab ; 297(3): E676-84, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19531635

RESUMO

The corpus luteum (CL) plays a central role in the maintenance of pregnancy in rodents, mainly by secreting progesterone. Female mice lacking prolactin (PRL) receptor (R) are sterile due to a failure of embryo implantation, which is a consequence of decreased luteinizing hormone (LH) receptor expression in the CL and inadequate levels of progesterone. We attempted to treat PRLR(-/-) females with human chorionic gonadotropin (hCG) and showed a de novo expression of LHR mRNA in the corpora lutea. Binding analysis confirmed that the LHR in hCG-treated PRLR(-/-) animals was functional. This was accompanied with increased expression of steroidogenic enzymes involved in progesterone synthesis. Despite these effects, no embryo implantation was observed because of high expression of 20alpha-hydroxysteroid dehydrogenase. To better appreciate the molecular mechanisms underlying maintenance of the CL, a series of mRNA expression-profiling experiments was performed on isolated corpora lutea of PRLR(-/-) and hCG-treated PRLR(-/-) mice. This approach revealed several novel candidate genes with potentially pivotal roles in ovarian function, among them, p27, VE-cadherin, Pten, and sFRP-4, a member of the Wnt/frizzled family. This study showed the differential role of PRL and LH in CL function and identified new targets of these hormones in luteal cells.


Assuntos
Manutenção do Corpo Lúteo/genética , Regulação da Expressão Gênica , Hormônio Luteinizante/fisiologia , Prolactina/fisiologia , Animais , Gonadotropina Coriônica/farmacologia , Corpo Lúteo/efeitos dos fármacos , Corpo Lúteo/metabolismo , Corpo Lúteo/fisiologia , Manutenção do Corpo Lúteo/sangue , Manutenção do Corpo Lúteo/efeitos dos fármacos , Manutenção do Corpo Lúteo/metabolismo , Feminino , Fertilidade/efeitos dos fármacos , Fertilidade/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Hormônio Luteinizante/farmacologia , Masculino , Camundongos , Camundongos Knockout , Ovário/anatomia & histologia , Ovário/efeitos dos fármacos , Ovário/metabolismo , Gravidez , Progesterona/sangue , Prolactina/farmacologia , Receptores do LH/genética , Receptores do LH/metabolismo , Receptores da Prolactina/genética
11.
Reprod Domest Anim ; 43 Suppl 2: 104-12, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18638111

RESUMO

Conception rates of dairy cows are currently declining at an estimated 1% every year. Approximately, 35% of embryos fail to prevent luteolysis during the first three weeks of gestation. Interactions between the corpus luteum, endometrium and embryo are critical to the successful establishment of pregnancy and inadequacies will result in the mortality of the embryo. For example, as little as a one day delay in the post-ovulatory rise of progesterone has serious consequences for embryo development and survival. Recently, we found that LH support, degree of vascularization and luteal cell steroidogenic capacity were not the major factors responsible for this luteal inadequacy, but are nevertheless essential for luteal development and function. Progesterone acting on its receptor in the endometrium stimulates the production of endometrial secretions on which the free-living embryo is dependent. However, their exact composition and effects of inadequate progesterone remains to be determined. The embryo is recognized through its secretion of interferon tau (IFNT), which suppresses luteolytic pulses of prostaglandin F(2 alpha). In the cow, it is most likely that IFNT inhibits oxytocin receptor up-regulation directly and does not require the prior inhibition of oestrogen receptor alpha (ESR1). Unravelling the precise luteal-endometrium and embryo interactions is essential for us to understand pregnancy establishment and development of strategies to reverse the declining fertility of dairy cows.


Assuntos
Bovinos/fisiologia , Corpo Lúteo/fisiologia , Embrião de Mamíferos/fisiologia , Endométrio/fisiologia , Proteínas da Gravidez/metabolismo , Prenhez/fisiologia , Animais , Bovinos/metabolismo , Corpo Lúteo/metabolismo , Manutenção do Corpo Lúteo/metabolismo , Manutenção do Corpo Lúteo/fisiologia , Embrião de Mamíferos/metabolismo , Endométrio/metabolismo , Feminino , Gravidez , Prenhez/metabolismo
12.
Reproduction ; 133(1): 187-96, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17244745

RESUMO

Peroxisome proliferator-activated receptor gamma (PPARgamma) has been shown to stimulate progesterone production by bovine luteal cells. We previously reported higher expression of PPARgamma in old compared with new luteal tissue in the rat. The following studies were conducted to determine the role of PPARgamma in rat corpora lutea (CL) and test the hypothesis that PPARgamma plays a role in the metabolism of progesterone and/or luteal lifespan. Ovaries were removed from naturally cycling rats throughout the estrous cycle, and pseudopregnant rats. mRNA for PPARgamma and P450 side-chain cleavage (SCC) was localized in luteal tissue by in situ hybridization, and protein corresponding to PPARgamma and macrophages identified by immunohistochemistry. Luteal tissue was cultured with agonists (ciglitazone, prostaglandin J2) or an antagonist (GW-9662) of PPARgamma. Progesterone was measured in media by RIA and levels of mRNA for 20alpha-hydroxysteriod dehydrogenase (HSD) and bcl-2 were measured in luteal tissue after culture by RT-PCR. An inverse relationship existed between the expression of mRNA for SCC and PPARgamma. There was no effect of PPARgamma agonists or the antagonist on luteal progesterone production in vitro, or levels of mRNA for 20alpha-HSD. PPARgamma protein was localized to the nuclei of luteal cells and did not correspond with the presence of macrophages. In new CL, ciglitazone decreased mRNA for bcl-2 on proestrus, estrus, and metestrus. Interestingly, GW-9662 also decreased mRNA for bcl-2 on proestrus and diestrus in old and new CL, and on metestrus in new CL. These data indicate that PPARgamma is not a major player in luteal progesterone production or metabolism but may be involved in regulating luteal lifespan.


Assuntos
Manutenção do Corpo Lúteo/metabolismo , Corpo Lúteo/metabolismo , PPAR gama/fisiologia , Anilidas/farmacologia , Animais , Núcleo Celular/química , Núcleo Celular/metabolismo , Enzima de Clivagem da Cadeia Lateral do Colesterol/genética , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Feminino , Imuno-Histoquímica , Hibridização In Situ , PPAR gama/antagonistas & inibidores , PPAR gama/genética , Gravidez , Prostaglandina D2/análogos & derivados , Prostaglandina D2/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Pseudogravidez , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tiazolidinedionas/farmacologia , Técnicas de Cultura de Tecidos
13.
Reproduction ; 133(1): 197-205, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17244746

RESUMO

It has been shown that hypothyroidism in the rat produces a prolongation of pregnancy associated with a delay in the fall of circulating progesterone (P4) at term. The aim of the present work is to determine whether the delayed P4 decline in hypothyroid mother rats is due to a retarded induction of P4 degradation to 20alphaOH P4 or to a stimulation of its synthesis, and to investigate the possible mechanisms that may underlie the altered luteal function. We determined by RIA the circulating profile of the hormones (TSH, PRL, LH, P4, PGF2alpha, and PGE2) involved in luteal regulation at the end of pregnancy and, by semiquantitative RT-PCR, the expression of factors involved in P4 synthesis (CytP450scc, StAR, 3betaHSD, PRLR) and metabolism (20alphaHSD, PGF2alphaR, iNOS and COX2). Our results show that the delay in P4 decline and parturition is the resultant of retarded luteal regression, caused by a combination of decreases in luteolytic factors, mainly luteal PGF2alpha, iNOS mRNA expression and also circulating LH, and increased synthesis or action of luteotrophic factors, such as luteal and circulating PGE2 and circulating PRL. All these changes may be direct causes of the decreased 20alphaHSD mRNA and protein (measured by western blot analysis) expression, which in the presence of unchanged expression of the factors involved in P4 synthesis results in elevated luteal and circulating P4 that prolonged pregnancy and also may favor longer survival of the corpus luteum.


Assuntos
Manutenção do Corpo Lúteo/metabolismo , Hipotireoidismo/metabolismo , Progesterona/sangue , 20-Hidroxiesteroide Desidrogenases/genética , 20-Hidroxiesteroide Desidrogenases/metabolismo , Animais , Western Blotting , Enzima de Clivagem da Cadeia Lateral do Colesterol/genética , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Corpo Lúteo/metabolismo , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Feminino , Expressão Gênica , Hormônio Luteinizante/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Gravidez , RNA Mensageiro/análise , Ratos , Ratos Wistar , Receptores de Prostaglandina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
Mol Reprod Dev ; 74(6): 730-9, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17120307

RESUMO

The present study was undertaken to determine the expression of vascular endothelial growth factor (VEGF) and its receptors, fms-like tyrosine kinase (Flt-1) and fetal liver kinase-1/kinase insert domain-containing receptor (Flk-1/KDR), in the porcine corpus luteum (CL) during the estrous cycle and early pregnancy. Immunohistochemical studies localized proteins of VEGF ligand-receptor system in the cytoplasm of luteal cells and in some blood vessels. Western blot analysis revealed significantly higher levels of VEGF protein during early and mid-luteal phase (vs. late luteal phase; P<0.001 and P<0.01, respectively). Quantification of VEGF mRNA in the CL showed increased mRNA levels during entire luteal phase (vs. Days 16-17; P<0.05). Expression of Flt-1 protein remained high during luteal phase (P<0.001), but the mRNA levels tended to increase from the early to the late luteal phase. Elevated protein expression of Flk-1/KDR was found in the mid-luteal phase (vs. Days 16-17; P<0.05). However, induction of Flk-1/KDR mRNA expression occurred earlier, in early luteal phase. The lowest VEGF, Flt-1 and Flk-1/KDR mRNA and protein levels were observed in regressed CL (P<0.001). During pregnancy, VEGF, Flt-1 and Flk-1/KDR mRNA and protein expression was comparable to the mid-luteal phase. In conclusion, the present study has demonstrated dynamic expression of VEGF and its receptors in the porcine CL during the estrous cycle and early pregnancy. These data suggest that the VEGF ligand-receptor system may play an important role in the development and maintenance of the CL in pigs.


Assuntos
Corpo Lúteo/metabolismo , Ciclo Estral/genética , Prenhez , Suínos/genética , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Animais , Manutenção do Corpo Lúteo/genética , Manutenção do Corpo Lúteo/metabolismo , Ciclo Estral/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Idade Gestacional , Gravidez , RNA Mensageiro/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
15.
Gynecol Obstet Invest ; 60(2): 67-74, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15785074

RESUMO

OBJECTIVE: To determine if luteal phase support improves the pregnancy rate in in vitro fertilization (IVF) cycles. DESIGN: A meta-analysis of randomized trials of luteal phase support was carried out with the main outcome measure being the pregnancy rate per cycle. RESULTS: Fifty-nine trials were evaluated. Eighteen trials met the inclusion criteria. Five main themes were identified: human chorionic gonadotropin (hCG) versus progesterone; progesterone versus progesterone and hCG; progesterone versus placebo; hCG versus placebo, and hCG versus progesterone versus no support. CONCLUSION: Luteal phase support is definitely indicated in IVF treatment cycles. This meta-analysis favored hCG above progesterone as luteal phase support with respect to pregnancy rates. Further prospective randomized trials are needed to determine a definite consensus with respect to the duration of luteal phase support in IVF cycles.


Assuntos
Gonadotropina Coriônica/farmacologia , Manutenção do Corpo Lúteo/efeitos dos fármacos , Fertilização in vitro/métodos , Progesterona/farmacologia , Manutenção do Corpo Lúteo/metabolismo , Feminino , Humanos , Gravidez , Taxa de Gravidez , Ensaios Clínicos Controlados Aleatórios como Assunto
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