Sengupta Anamika, Sridaran Rajagopala
Department of Physiology, Morehouse School of Medicine, 720 Westview Drive S.W., Atlanta, GA 30310-1495, USA.
J Histochem Cytochem. 2008 Jan;56(1):25-31. doi: 10.1369/jhc.7A7289.2007. Epub 2007 Sep 17.
A recent study from our laboratory has shown cellular and ultrastructural distribution of the gonadotropin-releasing hormone (GnRH) and the relative expression of its mRNA in the rat oviduct during the postimplantation period of pregnancy (days 7, 9, 16, and 20). To determine the possible autocrine/paracrine involvement of the oviductal GnRH during pregnancy in rats, the present investigation aims at the study of the relative expression of GnRH receptor (GnRHR) mRNA by real-time PCR followed by immunolocalization of the peptide in the oviduct during pregnancy. Semiquantitative analysis of the oviductal GnRHR expression by Western blot was done by densitometry of the signal intensity. Our results indicate the expression of GnRHR mRNA in the rat oviduct throughout the postimplantation period of pregnancy with no significant difference in expression between the selected time points. Immunoreactive GnRHR peptide was localized predominantly in the cytoplasm of the luminal epithelial cells, with less expression in the cytoplasm of the stromal cells and the smooth muscles throughout the oviduct. Signal intensity of GnRHR was significantly lower during day 16 when compared to days 7 and 20. These results, for the first time, support the transcription of GnRHR mRNA and its translation to protein in the rat oviduct throughout the postimplantation period of pregnancy. The lower protein content of GnRHR by day 16 may be indicative of ligand-induced downregulation of the receptor expression. The present investigation thus strengthens our previously postulated hypothesis regarding the receptor-mediated autocrine/paracrine role of oviductal GnRH during pregnancy in rats.
我们实验室最近的一项研究显示了妊娠植入后时期(第7、9、16和20天)大鼠输卵管中促性腺激素释放激素(GnRH)的细胞和超微结构分布及其mRNA的相对表达。为了确定大鼠妊娠期间输卵管GnRH可能的自分泌/旁分泌作用,本研究旨在通过实时PCR研究GnRH受体(GnRHR)mRNA的相对表达,随后对妊娠期间输卵管中的该肽进行免疫定位。通过对信号强度进行光密度测定,采用蛋白质印迹法对输卵管GnRHR表达进行半定量分析。我们的结果表明,在妊娠植入后整个时期大鼠输卵管中均有GnRHR mRNA表达,所选时间点之间的表达无显著差异。免疫反应性GnRHR肽主要定位于腔上皮细胞的细胞质中,在整个输卵管的基质细胞和平滑肌细胞质中的表达较少。与第7天和第20天相比,第16天时GnRHR的信号强度显著降低。这些结果首次支持了在妊娠植入后整个时期大鼠输卵管中GnRHR mRNA的转录及其向蛋白质的翻译。第16天时GnRHR较低的蛋白质含量可能表明配体诱导的受体表达下调。因此,本研究强化了我们之前提出的关于大鼠妊娠期间输卵管GnRH受体介导自分泌/旁分泌作用的假设。