热应激下公兔睾丸组织形态和精液转录组分析

doi:10.11843/j.issn.0366-6964.2023.11.020

Abstract

Abstract: The study aimed to understand the morphological changes of testicular tissue and responding gene profile of male rabbits in heat stress environment to reveal the mechanism of heat stress of the decline of semen quality of male rabbits. In this study, six 8-month-old New Zealand male rabbits with the same feeding level and similar body condition and weight were selected. By establishing a heat stress model, 3 rabbits in the heat stress group (August, HS ) and 3 rabbits in the non-heat stress group (May, NHS ) were obtained. The testis and semen of the two groups were collected, respectively. HE staining was used to compare the differences in testicular tissue structure under different conditions, and Illumina Hiseq high-throughput sequencing technology was used to analyze the transcriptome sequencing of sperm. GO and KEGG enrichment analysis of differentially expressed genes between the two groups of samples were performed by bioinformatics methods, and the transcriptome sequencing data were verified by RT-qPCR method. The results of testicular tissue sections of male rabbits showed that, compared with the NHS group, the central part in testis of the HS group showed obvious vacuolization and tearing, the spermatogenic epithelium was thinner, the damage was significant, and the number of sperm cells decreased. With log₂FoldChange>1 and P<0.05 as the threshold, a total of 1 676 differentially expressed genes related to sperm heat stress response were screened, including ATP5MC1, MDH2, ALDH7A1, GAPDHS, PRPS1, etc. Among them, 894 genes were up-regulated and 782 genes were down-regulated. GO analysis result showed that differentially expressed genes was enriched in stress response, protein metabolic process, catalytic activity and other related items, and KEGG result showed that differentially expressed genes was enriched in MAPK, PI3K-AkT, Toll-like receptor, Ras and other signaling pathways. RT-qPCR verified the expression levels of ATP5MC1, MDH2, PRPS1, ALDH7A1 and GAPDHS, which were consistent with the trend of transcriptome results. In summary, heat stress can regulate the expression of spermatogenesis-related genes and affect related signaling pathways and biological functions. The results of this study provide clues for the molecular regulation mechanism of spermato-genesis and provide a theoretical basis for the breeding of rabbits under high temperature environment.

Key words: heat stress, male rabbit, sperm, RNA-Seq, differentially expressed genes

CLC Number:

S814
S829.1

CAI Jiawei, ZHANG Chen, JIN Rongshuai, BAO Zhiyuan, ZHANG Xiyu, WANG Fan, ZHAI Pin, ZHAO Bohao, CHEN Yang, TANG Xianwei, WU Xinsheng. Analysis of Testicular Tissue Morphology and Semen Transcriptome of Male Rabbits under Heat Stress[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4653-4663.

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Analysis of Testicular Tissue Morphology and Semen Transcriptome of Male Rabbits under Heat Stress

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