National Engineering Laboratory for Animal Breeding & Key Laboratory of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, China Agricultural University, 100193, Beijing, People's Republic of China.
Mol Biol Rep. 2014 Mar;41(3):1597-606. doi: 10.1007/s11033-013-3006-4. Epub 2014 Jan 10.
There is accumulating evidence that leptin may be directly involved in mammalian reproduction, however, the potential role of obesity gene/obesity gene long form receptor (ob/ob-Rb) system in porcine implantation is poorly understood. To further confirm this role, mRNA and protein expression of ob/ob-Rb in implantation site and inter-implantation sites of porcine uterus on pregnancy day 13, 18 and 24 were compared in this study. Ob mRNA level went up with the advance of pregnancy and was higher in implantation site than inter-implantation site (P < 0.05). But ob-Rb mRNA, which was negative-regulated by leptin, went down with the advance of pregnancy and lessened in implantation site compared with inter-implantation site (P < 0.05). During the three implantation phase, leptin protein peaked at day 18 pregnancy (P < 0.05) and leptin protein at implantation site were always higher than inter-implantation site (P < 0.05). The higher ob-Rb protein in implantation site compared with inter-implantation site (P < 0.05) only appeared at day 18 pregnancy. Localization of ob/ob-Rb protein in porcine uterus was assayed using immunohistochemistry and found that ob/ob-Rb protein mainly located in luminal epithelium and glandular epithelium in pregnant pigs, but distinct immune-staining of leptin also detected in stroma in non-pregnancy porcine uterus except for luminal epithelium and glandular epithelium. In conclusion, the peak of leptin and the peak of ob-Rb protein in implantation site specifically appeared on day 18 pregnancy of pig. Another funning discovery is ob-Rb mRNA in porcine endometrium was mainly negative-regulated by leptin. The space-time difference of gene and protein expression for ob/ob-Rb confirmed ob/ob-Rb system role as delicate regulator of porcine implantation process.
越来越多的证据表明,瘦素可能直接参与哺乳动物的生殖过程,然而,肥胖基因/肥胖基因长型受体(ob/ob-Rb)系统在猪植入中的潜在作用还知之甚少。为了进一步证实这一作用,本研究比较了妊娠第 13、18 和 24 天猪子宫植入部位和植入部位之间 ob/ob-Rb 的 mRNA 和蛋白表达。ob mRNA 水平随着妊娠的进展而升高,在植入部位高于植入部位(P < 0.05)。但是,瘦素负调控的 ob-Rb mRNA 随着妊娠的进展而下降,在植入部位比植入部位少(P < 0.05)。在三个植入阶段,瘦素蛋白在妊娠第 18 天达到峰值(P < 0.05),植入部位的瘦素蛋白始终高于植入部位(P < 0.05)。植入部位的 ob-Rb 蛋白高于植入部位(P < 0.05)仅出现在妊娠第 18 天。用免疫组织化学法检测猪子宫中 ob/ob-Rb 蛋白的定位,发现 ob/ob-Rb 蛋白主要位于怀孕猪的腔上皮和腺上皮,而在非怀孕猪的子宫基质中除腔上皮和腺上皮外,还检测到瘦素的明显免疫染色。总之,瘦素和 ob-Rb 蛋白在植入部位的峰值专门出现在猪妊娠的第 18 天。另一个有趣的发现是,猪子宫内膜中 ob-Rb mRNA 主要受瘦素负调控。ob/ob-Rb 的基因和蛋白表达的时空差异证实了 ob/ob-Rb 系统作为猪植入过程精细调节剂的作用。