长线程和短线程RNA-seq揭示保山猪睾丸PICK1的转录调控特征

IF 1.6 4区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Animal Reproduction Pub Date : 2024-10-04 eCollection Date: 2024-01-01 DOI:10.1590/1984-3143-AR2024-0047
Xia Zhang, Hailong Huo, Guowen Fu, Changyao Li, Wan Lin, Hongmei Dai, Xuemin Xi, Lan Zhai, Qingting Yuan, Guiying Zhao, Jinlong Huo
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引用次数: 0

摘要

PICK1 在哺乳动物的精子发生过程中发挥着至关重要的作用。在此,我们整合了单分子长读程和短读程测序技术,全面研究了PICK1在成年保山猪(BS)睾丸中的表达模式。我们鉴定了PICK1最重要的转录本ENSSSCT00000000120,获得了其全长1254 bp的编码序列(CDS)。基因结构分析将 PICK1 定位在猪的第 5 号染色体上,共有 14 个外显子。蛋白质结构分析表明,PICK1 由 417 个氨基酸组成,包含两个保守结构域 PDZ 和 BAR_PICK1。系统发育分析强调了 PICK1 在不同哺乳动物物种中的进化保守性和同源性。对蛋白质相互作用网络、KEGG和GO通路的评估表明,PICK1与50个蛋白质相互作用,主要涉及谷氨酸能突触、苯丙胺成瘾、神经活性配体-受体相互作用、多巴胺能突触和突触小泡再循环,并且PICK1与DLG4和TBC1D20表现出显著的相关性。功能注释发现,PICK1参与了9个GOs,包括7个细胞成分和2个分子功能。ceRNA网络分析表明,BS PICK1受7个miRNA靶标调控。此外,对 15 个组织进行的 qPCR 表达分析表明,PICK1 在球尿道腺和睾丸中高表达。ST(Swine Tesits)细胞的亚细胞定位分析表明,PICK1在细胞质内显著定位。总之,我们的研究结果为 PICK1 在 BS 繁殖中的作用提供了新的线索,为进一步研究 PICK1 的功能奠定了基础。
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Long-read and short-read RNA-seq reveal the transcriptional regulation characteristics of PICK1 in Baoshan pig testis.

PICK1 plays a crucial role in mammalian spermatogenesis. Here, we integrated single-molecule long-read and short-read sequencing to comprehensively examine PICK1 expression patterns in adult Baoshan pig (BS) testes. We identified the most important transcript ENSSSCT00000000120 of PICK1, obtaining its full-length coding sequence (CDS) spanning 1254 bp. Gene structure analysis located PICK1 on pig chromosome 5 with 14 exons. Protein structure analysis reflected that PICK1 consisted of 417 amino acids containing two conserved domains, PDZ and BAR_PICK1. Phylogenetic analysis underscored the evolutionary conservation and homology of PICK1 across different mammalian species. Evaluation of protein interaction network, KEGG, and GO pathways implied that interacted with 50 proteins, predominantly involved in glutamatergic synapses, amphetamine addiction, neuroactive ligand-receptor interactions, dopaminergic synapses, and synaptic vesicle recycling, and PICK1 exhibited significant correlation with DLG4 and TBC1D20. Functional annotation identified that PICK1 was involved in 9 GOs, including seven cellular components and two molecular functions. ceRNA network analysis suggested BS PICK1 was regulated by seven miRNA targets. Moreover, qPCR expression analysis across 15 tissues highlighted that PICK1 was highly expressed in the bulbourethral gland and testis. Subcellular localization analysis in ST (Swine Tesits) cells demonstrated that PICK1 significantly localized within the cytoplasm. Overall, our findings shed new light on PICK1's role in BS reproduction, providing a foundation for further functional studies of PICK1.

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来源期刊
Animal Reproduction
Animal Reproduction AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
2.30
自引率
11.80%
发文量
49
审稿时长
70 days
期刊介绍: Animal Reproduction (AR) publishes original scientific papers and invited literature reviews, in the form of Basic Research, Biotechnology, Applied Research and Review Articles, with the goal of contributing to a better understanding of phenomena related to animal reproduction. The scope of the journal applies to students, researchers and practitioners in the fields of veterinary, biology and animal science, also being of interest to practitioners of human medicine. Animal Reproduction Journal is the official organ of the Brazilian College of Animal Reproduction in Brazil.
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