Genome Analysis Laboratory, Animal Genetics Division, Indian Veterinary Research Institute, Izatnagar, India.
Anim Reprod Sci. 2012 Aug;133(3-4):159-68. doi: 10.1016/j.anireprosci.2012.06.023. Epub 2012 Jul 6.
Interferon stimulated gene 15 (ISG15), one of the several proteins induced by conceptus derived Type I and/or a Type II interferon (IFN), is implicated as an important factor in determining the uterine receptivity and conceptus development. However, presence as well as specific role of the ISG15 in buffalo (Bubalus bubalis) reproduction is yet to be elucidated. In the present study, both genomic and cDNA sequences of bubaline (bu) ISG15 were cloned and investigated for its expression in different tissues of female reproductive tract of buffalo. Sequence analysis revealed 100% identity among the genomic sequences (1014 bp) of buISG15 from three different breeds of buffalo (viz., Murrah: Acc. No. DQ118137, Mehsana: Acc. No. DQ118138, and Nagpuri: Acc. No. DQ118136) and cDNAs (Acc. Nos. HM543268-HM543270). As in cattle, the buISG15 was comprised of two exons of 57 bp and 520 bp encoding a peptide of 154 amino acids. Moreover, the buISG15 cDNA sequence exhibited 98.3% and 98.5% identity with that of taurine and indicine cattle, respectively. Subsequent reverse transcription PCR analysis revealed expression of the buISG15 in the uterine endometrium, corpus luteum (CL), corpus hemorrhagicum and oviduct. Quantitative Real Time PCR (RTqPCR) analysis also confirmed the constitutive expression of the buISG15 in the uterine endometrium during different stages (i.e. estrus, diestrus and proestrus) of estrous cycle and also during early (∼d 30-40) pregnancy. Western blot analysis of the endometrial extract from both estrous cyclic as well as pregnant buffalo demonstrated the presence of only conjugated ISG15 which was >40 kDa. ISG15 mRNA and immune-reactive proteins were localized in the stromal as well as glandular epithelial cells of the uterine endometrium of estrous cyclic as well as pregnant buffalo. However, there was no significant difference in amount of ISG15 mRNA across the different reproductive phases. To conclude, this study will be helpful for the further understanding of the roles of the ISG15 in pregnancy of buffalo cows.
干扰素刺激基因 15（ISG15）是由胚胎衍生的 I 型和/或 II 型干扰素（IFN）诱导的几种蛋白质之一，被认为是决定子宫容受性和胚胎发育的重要因素。然而，水牛（Bubalus bubalis）中 ISG15 的存在及其特定作用仍有待阐明。在本研究中，克隆了水牛的基因组和 cDNA 序列，并研究了其在水牛生殖道不同组织中的表达。序列分析显示，来自三个不同品种（即 Murrah：Acc. No. DQ118137、Mehsana：Acc. No. DQ118138 和 Nagpuri：Acc. No. DQ118136）的 buISG15 基因组序列（1014 bp）之间具有 100%的同一性，cDNAs（Acc. Nos. HM543268-HM543270）也是如此。与牛一样，buISG15 由 57 bp 和 520 bp 的两个外显子组成，编码 154 个氨基酸的肽。此外，buISG15 cDNA 序列与牛和印度牛的分别具有 98.3%和 98.5%的同一性。随后的反转录 PCR 分析显示，buISG15 在子宫子宫内膜、黄体（CL）、出血体和输卵管中表达。定量实时 PCR（RTqPCR）分析也证实了 buISG15 在发情周期的不同阶段（即发情、间情期和发情前期）以及早期（约 d 30-40）妊娠期间在子宫子宫内膜中的组成型表达。来自发情周期和妊娠水牛的子宫内膜提取物的 Western blot 分析表明，只有共轭 ISG15 存在，其分子量>40 kDa。ISG15 mRNA 和免疫反应性蛋白定位于发情周期和妊娠水牛的子宫子宫内膜的基质和腺上皮细胞中。然而，在不同的生殖阶段，ISG15 mRNA 的量没有显著差异。总之，这项研究将有助于进一步了解 ISG15 在水牛妊娠中的作用。
