摘要
目的CatSper1是精子特定的电压门控Ca^(2+)通道蛋白,在精子生成和受精过程中具有重要作用。本研究旨在分析CatSper1基因在版纳微型猪近交系(BMI)的表达调控模式、序列特征与潜在的生物学功能。方法取成年BMI公猪睾丸进行转录组测序;采用RT-PCR技术克隆CatSper1的完整编码序列;分析CatSper1的序列、结构特征及相互作用蛋白;利用转录组测序数据构建CatSper1的lncRNA和miRNA调控网络,并进行GO功能注释。结果RNA-seq获得CatSper1基因的平均表达量和平均TPM值分别为2817.5和33.6。CatSper1 CDS区全长为2166 bp(GenBank登录号:OK042306),编码721个氨基酸,含有233个氨基酸残基组成的Ion_trans保守结构域以及与精子生成相关的保守跨膜螺旋结构。CatSper1蛋白与10个雄性育性相关蛋白质存在相互作用,尤其与该基因家族成员CatSper2-4蛋白互作最为紧密。CatSper1基因受10个miRNAs靶向调控,分别有16个和14个lncRNAs与CatSper1竞争性结合ssc-miR-1343和ssc-miR-744。GO注释发现CatSper1基因在分子功能(MF)、生物学过程(BP)、细胞成分(CC)等方面均具有重要功能。结论本研究报道了CatSper1在BMI睾丸中的表达,基因的分子结构特征和表达调控网络,为深入研究CatSper1基因在BMI精子发生、精子获能和顶体反应等重要生物学过程中的功能奠定了基础。
Objective CatSper1 is a sperm-specific voltage-gated Ca^(2+) channel protein that plays an essential role in spermatogenesis and fertilization.We analyzed the expression regulation pattern,sequence characteristics,and potential biological function of CatSper1 gene of Banna mini-pig inbred(BMI)line.Methods The testes of adult BMI boars were used for RNA sequencing(RNA-seq),and the complete coding sequence of CatSper1 was cloned using reverse-transcription polymerase chain reaction.The sequence,structural characteristics,and interacting proteins of CatSper1 were analyzed.The lncRNA and miRNA regulatory network of CatSper1 was constructed,and gene ontology function annotations were carried out using RNA-seq data.Results The average expression level and transcripts per million of CatSper1 obtained by RNA-seq was 2817.5 and 33.6,respectively.The full-length coding sequence of CatSper1 was 2166 bp(GenBank accession number:OK042306),encoding 721 amino acids.CatSper1 protein contained an Ion_trans conserved domain of 233 amino acid residues and a conserved transmembrane helix structure related to spermatogenesis.CatSper1 protein interacted with 10 proteins related to male reproduction,especially CatSper2-4 of the CatSper family.Ten miRNAs regulated CatSper1 gene by a targeted mode,and 16 and 14 lncRNAs completed with CatSper1 for binding to ssc-miR-1343 and ssc-miR-744,respectively.The gene ontology annotation result indicated that the CatSper1 gene plays important roles in molecular functions,biological processes,and cellular components.Conclusions This study reported the expression CatSper1 in BMI testis,the molecular structure characteristics and expression regulatory network.The fundings will lay groundwork for further research of CatSper1 function in important biological processes such as spermatogenesis,sperm capacitation and acrosome reaction in BMI line.
作者
张霞
王配
刘志朋
霍海龙
代红梅
赵筱
霍金龙
ZHANG Xia;WANG Pei;LIU Zhipeng;HUO Hailong;DAI Hongmei;ZHAO Xiao;HUO Jinlong(Department of Life Science,Lyuliang University,Lvliang 033001,China;College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201;Yunnan Vocational and Technical College of Agriculture,Kunming 650212)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2022年第5期619-628,共10页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金项目(32060733,31460580,31660637,31660650)
吕梁学院博士科研启动基金项目。
关键词
版纳微型猪近交系
阳离子通道精子相关蛋白1
转录组测序
精子生成
转录调控
Banna mini-pig inbred line(BMI)
cation channel sperm-associated protein 1(CatSper1)
RNA-seq
spermatogenesis
transcriptional regulation
