SPATA3基因克隆及其在牦牛和犏牛睾丸中的差异表达研究

doi:10.11843/j.issn.0366-6964.2020.02.009

Abstract

Abstract: The aim of this research was to clone the spermatogenesis associated 3 (SPATA3) gene in yak, identify its expression pattern in various tissues of yak, and in different developmental stages in yak and cattle-yak testis, respectively, explore the effects of SPATA3 on testicular development and sperm maturity, and provide reliable basis data for studying the mechanism of SPATA3 in spermatogenesis of yak. The samples of yak heart, liver, spleen, lung, kidney, small intestine, brain, ovary, uterus and testis were collected after slaughtering. The total RNAs in different samples were extracted. The coding sequence of SPATA3 gene was cloned and the expression of SPATA3 in different tissues was detected by RT-PCR. The structure and function of SPATA3 were analyzed by bioinformatics softwares. The mRNA expression of SPATA3 gene in testicular different development stages of yak and cattle-yak was compared by quantitative real-time PCR (RT-qPCR). Then, the localization and expression of SPATA3 in yak and cattle-yak testis were detected by immunohistochemistry. The results showed that the CDS region of SPATA3 gene was 693 bp, encoding 230 amino acids; It had high homology with SPATA3 sequence of cattle, wild yak and buffalo. SPATA3 gene was specifically expressed in testis tissue, and no expression was observed in other tissues of yak. The results of RT-qPCR showed that SPATA3 gene was expressed during testicular development of yak, and the expression level in testis of adulthood yak(4-5 years old) was extremely significantly higher than that of fetal period yak(5-6 months old, P<0.01). And its expression in testis in different developmental stages of yak were all extremely significantly higher than that of cattle-yak (P<0.01). The results of immunohistochemistry showed that SPATA3 was expressed in cytoplasm of round spermatid and elongating spermatid, and its expression level in testis of adulthood yak was extremely significantly higher than that in testis of adulthood cattle-yak (P<0.01). The above results show that SPATA3 is highly conserved during evolution, and it is differentially expressed in testis tissues of yak and cattle-yak. Its low expression may cause sperm formation disorder and participate in sperm maturation process, but the specific function should be researched further.

Key words: yak, cattle-yak, SPATA3, immunohistochemistry, spermatogenesis, expression

CLC Number:

S823.85

HUANG Xiangyue, XIONG Xianrong, ZHANG Lei, MA Hongcheng, HAI Zhuo, LI Jian. Cloning of SPATA3 Gene and Its Differential Expression Analysis in Yak and Cattle-yak Testis[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(2): 279-287.

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Cloning of SPATA3 Gene and Its Differential Expression Analysis in Yak and Cattle-yak Testis

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