Wang Yixin, Jin Long, Ma Jideng, Chen Li, Fu Yuhua, Long Keren, Hu Silu, Song Yang, Shang Dazhi, Tang Qianzi, Wang Xun, Li Xuewei, Li Mingzhou
Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.
Key Lab of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Asian-Australas J Anim Sci. 2018 Feb;31(2):189-197. doi: 10.5713/ajas.17.0518. Epub 2017 Oct 20.
Hemicastration is a unilateral orchiectomy to remove an injured testis, which can induce hormonal changes and compensatory hypertrophy of the remaining testis, and may influence spermatogenesis. However, the underlying molecular mechanisms are poorly understood. Here, we investigated the impact of hemicastration on remaining testicular function.
Prepubertal mice (age 24 days) were hemicastrated, and their growth was monitored until they reached physical maturity (age 72 days). Subsequently, we determined testis DNA methylation patterns using reduced representation bisulfite sequencing of normal and hemicastrated mice. Moreover, we profiled the testicular gene expression patterns by RNA sequencing (RNA-seq) to examine whether methylation changes affected gene expression in hemicastrated mice.
Hemicastration did not significantly affect growth or testosterone (p>0.05) compared with control. The genome-wide DNA methylation pattern of remaining testis suggested that substantial genes harbored differentially methylated regions (1,139) in gene bodies, which were enriched in process of protein binding and cell adhesion. Moreover, RNA-seq results indicated that 46 differentially expressed genes (DEGs) involved in meiotic cell cycle, synaptonemal complex assembly and spermatogenesis were upregulated in the hemicastration group, while 197 DEGs were downregulated, which were related to arachidonic acid metabolism. Integrative analysis revealed that proteasome 26S subunit ATPase 3 interacting protein gene, which encodes a protein crucial for homologous recombination in spermatocytes, exhibited promoter hypomethylation and higher expression level in hemicastrated mice.
Global profiling of DNA methylation and gene expression demonstrated that hemicastration-induced compensatory response maintained normal growth and testicular morphological structure in mice.
半去势是一种单侧睾丸切除术，用于切除受伤的睾丸，这会引发激素变化以及剩余睾丸的代偿性肥大，并可能影响精子发生。然而，其潜在的分子机制尚不清楚。在此，我们研究了半去势对剩余睾丸功能的影响。
对青春期前小鼠（24日龄）进行半去势，并监测其生长直至达到生理成熟（72日龄）。随后，我们使用正常和半去势小鼠的简化代表性亚硫酸氢盐测序来确定睾丸DNA甲基化模式。此外，我们通过RNA测序（RNA-seq）分析睾丸基因表达模式，以检查甲基化变化是否影响半去势小鼠的基因表达。
与对照组相比，半去势对生长或睾酮水平没有显著影响（p>0.05）。剩余睾丸的全基因组DNA甲基化模式表明，大量基因在基因体内含有差异甲基化区域（1139个），这些区域在蛋白质结合和细胞黏附过程中富集。此外，RNA-seq结果表明，半去势组中46个参与减数分裂细胞周期、联会复合体组装和精子发生的差异表达基因（DEG）上调，而197个DEG下调，这些下调基因与花生四烯酸代谢有关。综合分析显示，蛋白酶体26S亚基ATP酶3相互作用蛋白基因（该基因编码对精母细胞同源重组至关重要的一种蛋白质）在半去势小鼠中表现出启动子低甲基化和更高的表达水平。
DNA甲基化和基因表达的整体分析表明，半去势诱导的代偿反应维持了小鼠的正常生长和睾丸形态结构。
