Baatar Tergel, Song Dailing, Weng Yajuan, Wang Guoqing, Jin Liangyi, Guo Rui, Li Bei, Dugarjaviin Manglai
Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
Animals (Basel). 2025 Apr 11;15(8):1106. doi: 10.3390/ani15081106.
The epigenetic regulation of gene expression through the covalent modification of histones is crucial for developing germline cells. To study the regulatory role of alternative splicing (AS) of euchromatic histone lysine methyltransferase 2 (EHMT2/G9A) in spermatogenesis in Mongolian horses, this study first examines the localization of the gene in testicular support cells and then predicts the higher-order structures of sequences with and without AS. Two types of lentiviral vectors for overexpression were subsequently constructed for the gene, one with AS and one without, to infect support cells. The proliferation and activity of infected cells were measured using CCK8, and the differential expression of spermatogenesis-related genes in the two types of support cells was analyzed via qRT-PCR. We analyzed the expression of by immunofluorescence staining. was expressed in the nuclei of Sertoli cells. The expression of spermatogenesis-related genes was measured in the two types of cells. The results reveal that the expression levels of the , , , , , , , and genes in the AS group were greater than those in the control group. These results indicate that AS events in affect gene expression and thus affect spermatogenesis.
通过组蛋白的共价修饰对基因表达进行表观遗传调控对于生殖细胞的发育至关重要。为了研究常染色质组蛋白赖氨酸甲基转移酶2（EHMT2/G9A）的可变剪接（AS）在蒙古马精子发生中的调控作用，本研究首先检测了该基因在睾丸支持细胞中的定位，然后预测了有或无AS的序列的高级结构。随后构建了两种用于该基因过表达的慢病毒载体，一种带有AS，一种不带AS，以感染支持细胞。使用CCK8测量感染细胞的增殖和活性，并通过qRT-PCR分析两种类型支持细胞中精子发生相关基因的差异表达。我们通过免疫荧光染色分析了该基因的表达。该基因在支持细胞的细胞核中表达。在两种类型的细胞中测量了精子发生相关基因的表达。结果显示，AS组中该基因、、、、、、和基因的表达水平高于对照组。这些结果表明，该基因中的AS事件影响基因表达，从而影响精子发生。
