Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, and Key Laboratory of Reproduction and Genetics of Ningxia Hui Autonomous Region, Department of Anatomy, Histology and Embryology, School of Basic Medical Science, Ningxia Medical University, Yinchuan, 750004, China.
CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China.
Stem Cell Res Ther. 2019 Jul 29;10(1):227. doi: 10.1186/s13287-019-1335-5.
High temperature has a very adverse effect on mammalian spermatogenesis and eventually leads to sub- or infertility through either apoptosis or DNA damage. However, the direct effects of heat stress on the development of spermatogonial stem cells (SSCs) are still unknown because SSCs are rare in the testes.
In the present study, we first used in vitro-cultured SSCs to study the effect of heat shock treatment on SSC development. Then, we used RNA-Seq analysis to identify new genes or signalling pathways implicated in the heat stress response.
We found that 45 min of 43 °C heat shock treatment significantly inhibited the proliferation of SSCs 2 h after treatment but did not lead to apoptosis. In total, 17,822 genes were identified by RNA-Seq after SSC heat shock treatment. Among these genes, we found that 200 of them had significantly changed expression, with 173 upregulated and 27 downregulated genes. The number of differentially expressed genes in environmental information processing pathways was 37, which was the largest number. We screened the candidate JAK-STAT signalling pathway on the basis of inhibition of cell cycle progression and found that the JAK-STAT pathway was inhibited after heat shock treatment. The flow cytometry results further confirmed that heat stress caused S phase cycle arrest of SSCs.
Our results showed that heat shock treatment at 43 °C for 45 min significantly inhibited SSC self-renewal through S phase cell cycle arrest but not apoptosis.
高温对哺乳动物的精子发生有非常不利的影响，最终通过细胞凋亡或 DNA 损伤导致生育能力下降或不育。然而，由于精原干细胞（SSC）在睾丸中很少，因此热应激对 SSC 发育的直接影响尚不清楚。
在本研究中，我们首先使用体外培养的 SSC 来研究热休克处理对 SSC 发育的影响。然后，我们使用 RNA-Seq 分析来鉴定新的基因或信号通路参与热应激反应。
我们发现，43°C 的 45 分钟热休克处理在处理后 2 小时显著抑制了 SSC 的增殖，但不会导致细胞凋亡。通过 RNA-Seq 总共鉴定出 17822 个基因，其中 200 个基因的表达有显著变化，上调的基因有 173 个，下调的基因有 27 个。在环境信息处理途径中差异表达基因的数量为 37，是数量最多的。我们基于细胞周期进程的抑制筛选候选 JAK-STAT 信号通路，发现热休克处理后 JAK-STAT 通路被抑制。流式细胞术结果进一步证实，热应激导致 SSC 的 S 期细胞周期停滞。
我们的结果表明，43°C 热休克处理 45 分钟可通过 S 期细胞周期阻滞而不是细胞凋亡显著抑制 SSC 的自我更新。
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