山羊Wnt10b基因生物学特征及组织细胞表达模式分析

Analysis of Biological Characteristics and Tissue Cell Expression Pattern of Wnt10b Gene in Goat

为了获得山羊Wnt10b基因序列,并阐明其组织表达谱及在前体脂肪细胞和成肌细胞分化过程中的表达模式。本研究利用胶原酶消化法获得山羊皮下和肌内前体脂肪细胞,利用组织块法获得成肌细胞;采用RT-PCR方法克隆山羊Wnt10b基因序列,荧光定量PCR技术检测该基因在各组织、前体脂肪细胞与成肌细胞诱导分化过程中的表达情况。由分析可知,获得山羊Wnt10b基因序列1 232 bp(Gen Bank登陆号:KU950832),其中CDS为1 176 bp,5'UTR 44 bp和3'UTR 12 bp,编码391个氨基酸残基,山羊Wnt10b氨基酸序列与绵羊的氨基酸序列同源性达98%;Wnt10b m RNA在山羊脂肪组织中表达水平最高,极显著高于其他组织(p<0.01);Wnt10b m RNA随着山羊肌内前体脂肪细胞和成肌细胞的分化表达呈上升趋势,但其在皮下前体脂肪细胞中的表达模式却相反。结果表明,山羊Wnt10b基因在脂肪组织中表达水平最高,可能在脂肪沉积和脂代谢中发挥重要的调控作用,但在皮下和肌内前体脂肪细胞成脂分化的表达模式不同,提示该基因在山羊不同部位脂肪沉积中可能发挥不同的调控作用。

The aim of this study was to obtain the gene sequence of goat Wnt10 b, and clarify its expression pattern in different tissues and differentiation of preadipocyte and myoblast. The goat subcutaneous and intramuscular preadipocytes were obtained by collagenase digestion method, and myoblast was obtained by tissue block method. We used RT-PCR method to clone the gene sequence of goat Wnt10 b, and also used fluorescence quantitative PCR technique to detect the expression situation of Wnt10 b gene in the induced differentiation of each tissue, preadipocyte, and myoblast. The results showed that the gene sequence of goat Wnt10 b was 1 232 bp（Gen Bank accession number： KU950832）, containing 1 176 bp CDS, 44 bp 5＇ UTR and 12 bp 3＇ UTR, encoding391 amino acids. Homology of the amino acid sequence of goat Wnt10 b and sheep could reach 98%. The m RNA expression level of Wnt10 b was the highest in adipose tissue, which was extremely higher than other organizations（p〈0.01）. Its expression level gradually increased after the induction of intramuscular preadipocytes and myoblast,while it significantly decreased in subcutaneous preadipocytes. We found that the m RNA expression level of Wnt10 b was the highest in adipose tissue, so it might play an important regulatory role in fat deposition and lipid metabolism, but its expression patterns of subcutaneous and intramuscular preadipocyte were different, which suggested that Wnt10 b gene might play a different role in the fat deposition of different parts in goat.