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泛素在妊娠早期猪子宫内膜腔上皮中由干扰素生理诱导:用于差异显示筛选的全局逆转录聚合酶链反应互补脱氧核糖核酸替代核糖核酸。

Ubiquitin is physiologically induced by interferons in luminal epithelium of porcine uterine endometrium in early pregnancy: global RT-PCR cDNA in place of RNA for differential display screening.

作者信息

Chwetzoff S, d'Andrea S

机构信息

Institut National de la Recherche Agronomique, Laboratoire de Virologie et d'Immunologie Moleculaires, Jouy-en-Josas, France.

出版信息

FEBS Lett. 1997 Mar 24;405(2):148-52. doi: 10.1016/s0014-5793(97)00177-4.

Abstract

Early in the course of pregnancy, at the preimplantation stage, the pig embryo is likely to exert a paracrine effect on the tissue intended to receive it, via the secretion of interferons. Our observations show that trophoblastic interferons induce an increase of some mRNAs in the epithelial cells of the gilt endometrium, which would illustrate this phenomenon. The increase of four mRNAs, whose corresponding cDNAs are dD1, dD2, dD3 and dD4, has been examined in this study. The method used is similar to Northern blot analysis except that mRNAs in the blot are replaced by cDNAs produced from total cellular poly(A)+ mRNAs by global reverse-transcription polymerase chain reaction (RT-PCR). Northern blot hybridization requires a considerable quantity of starting material--which we estimate in this study to be several million porcine endometrium cells--whereas the RT-PCR-based method gives comparable results starting with only a few cells--about 200. Using this method, the differential nature of dD1, dD2, dD3 and dD4 was shown. dD2 and dD3 correspond to genes already identified as interferon-induced: the beta2-microglobulin and Finkel-Biskis-Reilly murine sarcoma virus-associated ubiquitously secreted protein (FAU). dD1 corresponds to a still unidentified gene. dD4 encodes for the porcine UbA52 ubiquitin. Up to now, the increase in ubiquitin mRNA as a result of interferon effect has not been reported and is discussed in view of recent publications.

摘要

在妊娠早期,即着床前阶段,猪胚胎可能通过分泌干扰素,对准备接纳它的组织产生旁分泌效应。我们的观察结果表明,滋养层干扰素可诱导后备母猪子宫内膜上皮细胞中某些mRNA的增加,这将说明这一现象。本研究检测了四种mRNA(其对应的cDNA分别为dD1、dD2、dD3和dD4)的增加情况。所采用的方法类似于Northern印迹分析,不同之处在于印迹中的mRNA被通过全细胞多聚腺苷酸加尾mRNA的整体逆转录聚合酶链反应(RT-PCR)产生的cDNA所取代。Northern印迹杂交需要大量的起始材料——在本研究中我们估计需要数百万个猪子宫内膜细胞——而基于RT-PCR的方法仅从少数细胞(约200个)开始就能得到类似的结果。使用这种方法,显示了dD1、dD2、dD3和dD4的差异性质。dD2和dD3对应于已被鉴定为干扰素诱导的基因:β2-微球蛋白和芬克尔-比斯克斯-赖利鼠肉瘤病毒相关的普遍分泌蛋白(FAU)。dD1对应于一个尚未鉴定的基因。dD4编码猪UbA52泛素。到目前为止,尚未有因干扰素作用导致泛素mRNA增加的报道,鉴于最近的出版物对此进行了讨论。

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