Lei Luo, Zhu Junxian, Chen Chen, Wang Yakun, Hong Xiaoyou, Liu Xiaoli, Yu Lingyun, Wei Chengqing, Chen Haigang, Liu Yihui, Li Ruiyang, Li Wei, Zhu Xinping
Key Laboratory of Tropical and Subtropical Fishery Resource Application and Cultivation, Ministry of Agriculture and Rural Affairs, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China.
Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China.
Animals (Basel). 2022 Jul 21;12(14):1858. doi: 10.3390/ani12141858.
Spats1 (spermatogenesis-associated, serinerich 1) has been characterized as a male-biased gene which acts an important role in the germ cell differentiation of mammals. Nevertheless, the function of Spats1 in the Chinese soft-shelled turtle (P. sinensis) has not yet been reported. To initially explore the expression of Spats1 in P. sinensis and its response to sex steroid treatment, we cloned the CDS of Spats1 for the first time and analyzed its expression profile in different tissues, including the testes in different seasons. The Spats1 cDNA fragment is 1201 base pairs (bp) in length and contains an open reading frame (ORF) of 849 bp, which codes for 283 amino acids. Spats1 mRNA was highly expressed in the testes (p < 0.01) and barely detectable in other tissues. In P. sinensis, the relative expression of Spats1 also responsive to seasonal changes in testis development. In summer (July) and autumn (October), Spats1 gene expression was significantly higher in the testes than in other seasons (p < 0.05). Spats1 mRNA was found to be specifically expressed in germ cells by chemical in situ hybridization (CISH), and it was mainly located in primary spermatocytes (Sc1), secondary spermatocytes (Sc2) and spermatozoa (St). Spats1 expression in embryos was not significantly changed after 17α-methyltestosterone (MT)and 17β-estradiol (E2) treatment. In adults, MT significantly induced Spats1 expression in male P. sinensis. However, the expression of Spats1 in testes was not responsive to E2 treatment. In addition, the expression of Spats1 in females was not affected by either MT or E2 treatment. These results imply that Spats1 is a male-specific expressed gene that is mainly regulated by MT and is closely linked to spermatogenesis and release in P. sinensis.
精子发生相关富含丝氨酸蛋白1（Spats1）已被鉴定为一种雄性偏向基因，在哺乳动物生殖细胞分化中起重要作用。然而，Spats1在中国鳖（中华鳖）中的功能尚未见报道。为初步探究Spats1在中华鳖中的表达及其对性类固醇处理的反应，我们首次克隆了Spats1的编码序列（CDS），并分析了其在不同组织中的表达谱，包括不同季节的睾丸。Spats1 cDNA片段长度为1201个碱基对（bp），包含一个849 bp的开放阅读框（ORF），编码283个氨基酸。Spats1 mRNA在睾丸中高表达（p < 0.01），在其他组织中几乎检测不到。在中华鳖中，Spats1的相对表达也对睾丸发育的季节变化有反应。在夏季（7月）和秋季（10月），睾丸中Spats1基因表达显著高于其他季节（p < 0.05）。通过化学原位杂交（CISH）发现Spats1 mRNA在生殖细胞中特异性表达，主要位于初级精母细胞（Sc1）、次级精母细胞（Sc2）和精子（St）中。17α-甲基睾酮（MT）和17β-雌二醇（E2）处理后，胚胎中Spats1的表达无显著变化。在成体中，MT显著诱导雄性中华鳖中Spats1的表达。然而，睾丸中Spats1的表达对E2处理无反应。此外，MT或E2处理均不影响雌性中华鳖中Spats1的表达。这些结果表明，Spats1是一个雄性特异性表达基因，主要受MT调控，与中华鳖的精子发生和释放密切相关。
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