牦牛瘦素基因克隆及其在肾周和皮下脂肪组织的表达和分布  

Cloning of Leptin Gene in Yak(Bos grunniens)and Its Expression and Distribution in Perirenal and Subcutaneous Adipose Tissues

作　　者：范伟峰 陈付菊[1] 赵宇田 李颖帮 童依然 陈杏娣 胡亚青 FANWei-Feng;CHEN Fu-Ju;ZHAO Yu-Tian;LI Ying-Bang;TONG Yi-Ran;CHEN Xing-Di;HU Ya-Qing(College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China;Agricultural Comprehensive Administrative Law Enforcement Supervision Bureau of Qinghai Province,Xining 810006,China;Bureau of Agriculture and Rural Affairs,Hualong County,Hualong 810900,China)

机构地区：[1]青海大学农牧学院,西宁810016 [2]青海省农业综合行政执法监督局,西宁810006 [3]化隆县农业农村和科技局,化隆810900

出　　处：《农业生物技术学报》2024年第10期2285-2292,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31960686)。

摘　　要：瘦素(leptin)在机体能量平衡及脂肪沉积方面具有重要作用,其在牦牛(Bos grunniens)能量代谢中的作用尚不明确。本研究采集1和30日龄的健康雄性犊牦牛肾周和皮下棕色及白色脂肪组织,通过RTPCR克隆leptin基因编码区序列(CDS),采用生物信息学方法分析其序列特征,并利用qRT-PCR和免疫组织化学(immunohistochemistry,IHC)染色方法检测其在牦牛肾周和皮下脂肪组织中的表达和分布。结果显示,牦牛leptin基因CDS序列(GenBank No.PP385937)长504 bp,编码167个氨基酸,与野牦牛(Bos mutus)的同源性可达99.01%,表明该基因在进化过程中较为保守。基因表达分析显示,与1日龄比较,30日龄牦牛肾周和皮下脂肪组织中leptin基因表达量显著增加(P<0.05)。免疫组织化学染色显示,1和30日龄牦牛的肾周和皮下棕色及白色脂肪细胞膜上均有leptin阳性染色,30日龄的染色强度均显著高于1日龄(P<0.05)。本研究为深入阐明leptin在牦牛寒冷适应中的作用机制提供了基础资料。Leptin plays an important role in the regulation of energy balance and fat deposition,but its role in energy metabolism in yaks(Bos grunniens)is not yet clear.This study collected perirenal and subcutaneous brown and white adipose tissues of 1-and 30-day-old healthy male yak calves,cloned the CDS of the leptin gene through RT-PCR,analyzed its sequence characteristics using bioinformatics methods,and detected its expression and distribution in yak perirenal and subcutaneous adipose tissues using qRT-PCR and immunohistochemistry(IHC)staining methods.The results showed that the CDS of yak leptin gene(GenBank No.PP385937)was 504 bp,encoding 167 amino acids,and the homology with wild yak(Bos mutus)reached 99.01%,indicating that the gene was relatively conserved during evolution.The gene expression analysis showed that the leptin expression in the perirenal and subcutaneous adipose tissues of 30-day-old yak were significantly higher than those of 1-day-old yak(P<0.05).The IHC showed that the leptin positive staining was observed on the cytomembrane of borwn and white adipocytes in perirenal and subcutaneous brown and white adipose tissues,and the leptin positive staining intensity of 30-day-old yak was significantly higher than that of 1-day-old yak(P<0.05).This study provides basic materials for further elucidating the mechanism of leptin in cold adaptation of yaks.

关 键 词：瘦素 犊牦牛 基因克隆 脂肪组织 基因表达 分布 

分 类 号：S852.1[农业科学—基础兽医学]