RESUMO
Mammals are important for balancing the natural ecosystem, but in the past few decades, several species have rapidly been entered under threatened category worldwide. The environmental changes, loss of natural habitats, human activities, and thereby stress are responsible for a gradual decline in reproductive outcome. Stress induces generation of reactive oxygen species (ROS). High physiological level of ROS drives abortive spontaneous egg activation (SEA), while beyond the physiological level causes oxidative stress (OS). The OS induces apoptosis and deteriorates egg quality that limits reproductive outcome. The reduced reproductive outcome is one of the major causes for gradual decline in population size of several mammalian species. Despite having several conservation programs, a gradual decline in species reproductive outcome and their population size is the serious concern for the existence of threatened mammalian species. Thus, it is important to identify and prevent the underlying causes responsible for abortive SEA, which could be an emerging problem for several mammalian species that are threatened or at the verge of extinction.
Assuntos
Apoptose , Mamíferos/metabolismo , Óvulo/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Reprodução , Animais , Feminino , MasculinoRESUMO
The present study was aimed to find out whether increased level of reactive oxygen species (ROS) particularity hydrogen peroxide (H2O2) could persuade postovulatory aging-mediated abortive spontaneous egg activation (SEA) in rat eggs cultured in vitro. For this purpose, ROS and H2O2 levels, mitochondria distribution and its membrane potential, p286-CaMK-II, Emi2, Thr-161 phophorylated cyclin-dependent protein kinase1 (Cdk1) as well as cyclin B1 levels, in vitro effects of 3-tert-butyl-4 hydroxy anisole (BHA), pentoxifylline and dibutyryl-adenosine 3',5'-cyclic monophosphate (db-cAMP) were analyzed during postovulatory aging-induced abortive SEA in vitro. Data of the present study suggest that postovulatory aging increased H2O2 levels, disturbed mitochondrial distribution pattern and mitochondrial membrane potential (MMP) in eggs. There was an significant increase of p286-CaMK-II level, while Emi2 level reduced significantly during egg aging in vitro. The reduced Emi2 level was associated with decreased Thr-161 phosphorylated cyclin-dependent kinase-1 (Cdk1) as well as cyclin B1 level in aged eggs that underwent abortive SEA. Further, supplementation of pentoxifylline, db-cAMP, and BHA protected postovulatory aging-mediated abortive SEA in concentration-dependent manner. These data suggest that postovulatory aging increased H2O2 levels, reduced MMP, and increased p286-CaMK-II. The increased p286-CaMK-II was associated with reduced Emi2 level and maturation-promoting factor levels during postovulatory aging-mediated abortive SEA. Drugs that elevate cAMP directly or indirectly and BHA protected postovulatory aging-mediated abortive SEA possibly by reducing ROS level in rat eggs cultured in vitro.
Assuntos
Senescência Celular , Peróxido de Hidrogênio/metabolismo , Fase Luteal , Oócitos/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Bucladesina/farmacologia , Hidroxianisol Butilado/farmacologia , Proteína Quinase CDC2 , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Ciclina B1/metabolismo , Quinases Ciclina-Dependentes/metabolismo , Proteínas F-Box/metabolismo , Feminino , Potencial da Membrana Mitocondrial , Mitocôndrias/metabolismo , Oócitos/efeitos dos fármacos , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Pentoxifilina/farmacologia , Fosforilação , Ratos EndogâmicosRESUMO
OBJECTIVE: The aim of this study was to determine whether an increase of intracellular nitric oxide (NO) level signals postovulatory aging-induced abortive spontaneous egg activation (SEA) in rats. METHODS: Freshly ovulated eggs (arrested at metaphase-II stage; M-II) were cultured in vitro for 3 hours to induce postovulatory egg aging. The morphological changes, inducible nitric oxide synthase (iNOS) expression, NO, cytosolic free Ca(2+), 3',5' cyclic guanosine monophosphate (cGMP), cell division cycle 25B (Cdc25B) and Wee1 levels, specific phosphorylation (pThr-14/Tyr-15) as well as total cyclin-dependent kinases-1 (Cdk1) (PSTAIRE) levels were analyzed. RESULTS: Postovulatory aging induced generation of NO possibly through an iNOS-mediated pathway. The increase in NO level was associated with augmented cytosolic free Ca(2+) as well as cGMP levels in aged eggs. A significant increase in Wee1 level and decrease of Cdc25B level were observed in aged eggs. An accumulation of phosphorylated Cdk1 (pThr-14/Tyr-15) level was observed in aged eggs, while total Cdk1 (PSTAIR) level remained unchanged. CONCLUSION: Our study demonstrates that generation of NO through an iNOS-mediated pathway increases cytosolic free Ca2+and cGMP levels. High levels of these signal molecules trigger the accumulation of phosphorylated Cdk1 in aged eggs. Thus, NO signals the accumulation of phosphorylated Cdk1 and induces postovulatory aging-induced abortive SEA in the rat.