RESUMO
Bovine interferon-tau (bIFN-tau) is secreted by the developing conceptus and initiates antiluteolytic events by interacting with uterine membrane receptors. We have identified three endometrial proteins (approximately 8, 16, and 28 kDa; P8, P16, and P28; respectively) that are secreted in response to recombinant (r) bIFN-tau. The objective of this study was to determine whether or not secretion of these proteins was a unique response to IFN-tau during early pregnancy. Three experiments were designed to examine secretion of endometrial proteins as a function of time in culture (0, 3, 6, 12, 18, 24 h), stage of the estrous cycle and pregnancy (Days 15, 18, 0/21), and dose of Type I IFN (0, 0.5, 5, and 25 nM; rbIFN-tau, rbIFN-alpha, and roIFN-tau). Endometrium was cultured for times specified with L-[3H]leusine to generate radiolabeled proteins. Secreted proteins were quantitated by using one-dimensional (1D)-PAGE, fluorography, and densitometry. Secretion of P8, P16, and P28 increased over time (p < 0.0001) in culture and in response to 25 nM rbIFN-tau (p < 0.05). Secretion of P8 in response to rbIFN-tau was higher (p < 0.05). Secretion of P8 in response to rbIFN-tau was higher (p < 0.0005) in endometrium collected from pregnant than nonpregnant heifers, but did not differ across the days examined. Although secretion of P8 was higher (p < 0.0001) in the presence than in the absence of rbIFN-tau, it was not affected by rbIFN-alpha.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Bovinos/metabolismo , Interferon Tipo I/farmacologia , Proteínas da Gravidez/farmacologia , Proteínas/metabolismo , Útero/metabolismo , Animais , Técnicas de Cultura , Relação Dose-Resposta a Droga , Eletroforese em Gel de Poliacrilamida , Estro/fisiologia , Feminino , Interferon Tipo I/administração & dosagem , Peso Molecular , Gravidez , Proteínas da Gravidez/administração & dosagem , Proteínas Recombinantes/farmacologia , Fatores de TempoRESUMO
Conceptus-derived interferon-tau (IFN-tau) has been implicated in the process of maternal recognition of pregnancy in the bovine. This type I IFN interacts with a uterine receptor complex to elicit secondary maternal responses, one of which is secretion of uterine proteins. We investigated the effects of pregnancy and recombinant (r) bovine (bo) IFN-tau (10(7) antiviral units/mg) on secreted proteins by cultured bovine endometrial explants. Explants were cultured for 24 h with [3H]leucine and dialyzed medium analyzed by one- and two-dimensional SDS polyacrylamide gel electrophoresis (PAGE) and fluorography. In one-dimensional PAGE experiments, endometrium representing early pregnancy (days 16-21) and the estrous cycle (days 16-19) was cultured in the presence of 5 nM rboIFN-tau and showed an increase (two- to five-fold) in secretion of 12- and 28-kD proteins. Further examination of these proteins by using two-dimensional PAGE indicated that the 12-kD protein was basic (pI > or = 7.5), whereas the 28-kD protein was acidic (pI approximately 5.0). Isoelectric focusing in the acidic range revealed that the 28-kD protein was composed of several isoelectric variants (pI 4.5-5.5). Although the functions of these secretory proteins are currently unknown, they serve as useful markers for IFN action and may act as secondary signals to protect the early developing conceptus.