Genome Analysis Laboratory, Animal Genetics Division, Indian Veterinary Research Institute, Izatnagar 243 122, India.
Anim Reprod Sci. 2010 Oct;122(1-2):133-41. doi: 10.1016/j.anireprosci.2010.08.005. Epub 2010 Aug 13.
Uterine serpins (SERPINA14) play important roles during pregnancy in the farm animals. In this study, we have cloned and characterized cDNA sequence encoding the bubaline SERPINA14. We also studied its spatio-temporal expression in the uterine endometrium. The bubaline SERPINA14 has an open reading frame of 1299bp. Itshares ∼90% identity with the SERPINA14 of other ruminant livestock species. Phylogenetically, bubaline SERPINA14 has been placed in the same clade that contained other mammalian homologues with a maximum identity to bovine SERPINA14. Using an anti-ovine monoclonal antibody, Western blot analysis of the uterine fluid of buffalo during the early stage of pregnancy confirmed the presence of SERPINA14 of about 48,000Da. The results of quantitative real time PCR (RT-qPCR) as well as in situ hybridization demonstrated a stage and tissue specific expression of bubaline SERPINA14. The level of SERPINA14 mRNA was low during stage I (Days 3-5), which increased (P<0.05) during stage II (Days 6-15) and then subsequently declined during stage III (Days 16-21) of the estrus cycle. During early pregnancy (Days ∼30 of gestation) the level of SERPINA14 mRNA was as high as that during stage II of the estrus cycle. The SERPINA14 mRNA was localized in the glandular epithelium. The differential spatio-temporal expression of SERPINA14 in the uterine endometrium of buffalo suggests its plausible important roles in reproduction.
绵羊丝氨酸蛋白酶抑制剂（SERPINA14）在农场动物的妊娠期间发挥着重要作用。在本研究中，我们克隆并鉴定了编码绵羊 SERPINA14 的 cDNA 序列。我们还研究了其在子宫子宫内膜中的时空表达。绵羊 SERPINA14 的开放阅读框为 1299bp。它与其他反刍家畜物种的 SERPINA14 具有约 90%的同源性。系统发育分析表明，绵羊 SERPINA14 与其他哺乳动物同源物位于同一分支，与牛 SERPINA14 的同源性最高。使用抗绵羊单克隆抗体，通过 Western blot 分析在妊娠早期的水牛子宫液中证实了约 48000Da 的 SERPINA14 的存在。定量实时 PCR（RT-qPCR）和原位杂交的结果表明，绵羊 SERPINA14 具有组织和阶段特异性表达。在发情周期的 I 期（第 3-5 天），SERPINA14 mRNA 的水平较低，在 II 期（第 6-15 天）增加（P<0.05），然后在 III 期（第 16-21 天）下降。在早期妊娠（妊娠约 30 天）时，SERPINA14 mRNA 的水平与发情周期的 II 期一样高。SERPINA14 mRNA 定位于腺上皮中。SERPINA14 在水牛子宫子宫内膜中的差异时空表达表明其在繁殖中可能具有重要作用。
