PDF(Pubmed)
Abstract：
BACKGROUND: Spermatogenesis is a temperature-sensitive process, and elevation in temperature hampers this process quickly and significantly. We studied the molecular effects of testicular heating on piRNAs and gene expression in rat testicular germ cells.
METHODS: We generated a cryptorchid rat model by displacing the testis from the scrotal sac (34 °C) to the abdominal area (37 °C) and sacrificed animals after 1 day, 3 days, and 5 days. Pachytene spermatocytes and round spermatids were purified using elutriation centrifugation and percoll gradient methods. We performed transcriptome sequencing in pachytene spermatocytes and round spermatids to identify differentially expressed piRNAs and their probable targets, i.e., TE transcripts and mRNAs.
RESULTS: As a result of heat stress, we observed significant upregulation of piRNAs and TE transcripts in testicular germ cells. In addition to this, piRNA biogenesis machinery and heat shock proteins (Hsp70 and Hsp90 family members) were upregulated. mRNAs have also been proposed as targets for piRNAs; therefore, we shortlisted certain piRNA-mRNA pairs with an inverse relationship of expression. We observed that in testicular heat stress, the heat shock proteins go hand-in-hand with the upregulation of piRNA biogenesis machinery. The dysregulation of piRNAs in heat-stressed germ cells, increased ping-pong activity, and disturbed expression of piRNA target transcripts suggest a connection between piRNAs, mRNAs, and TE transcripts.
CONCLUSIONS: In heat stress, piRNAs, piRNA machinery, and heat shock proteins are activated to deal with low levels of stress, which is followed by a rescue approach in prolonged stressaccompained by high TE activity to allow genetic mutations, perhaps for survival and adaptability.
METHODS: We generated a cryptorchid rat model by displacing the testis from the scrotal sac (34 °C) to the abdominal area (37 °C) and sacrificed animals after 1 day, 3 days, and 5 days. Pachytene spermatocytes and round spermatids were purified using elutriation centrifugation and percoll gradient methods. We performed transcriptome sequencing in pachytene spermatocytes and round spermatids to identify differentially expressed piRNAs and their probable targets, i.e., TE transcripts and mRNAs.
RESULTS: As a result of heat stress, we observed significant upregulation of piRNAs and TE transcripts in testicular germ cells. In addition to this, piRNA biogenesis machinery and heat shock proteins (Hsp70 and Hsp90 family members) were upregulated. mRNAs have also been proposed as targets for piRNAs; therefore, we shortlisted certain piRNA-mRNA pairs with an inverse relationship of expression. We observed that in testicular heat stress, the heat shock proteins go hand-in-hand with the upregulation of piRNA biogenesis machinery. The dysregulation of piRNAs in heat-stressed germ cells, increased ping-pong activity, and disturbed expression of piRNA target transcripts suggest a connection between piRNAs, mRNAs, and TE transcripts.
CONCLUSIONS: In heat stress, piRNAs, piRNA machinery, and heat shock proteins are activated to deal with low levels of stress, which is followed by a rescue approach in prolonged stressaccompained by high TE activity to allow genetic mutations, perhaps for survival and adaptability.
摘要：
背景：精子发生是一个对温度敏感的过程，温度的升高迅速而显著地阻碍了这一过程。我们研究了睾丸加热对大鼠睾丸生殖细胞中piRNA和基因表达的分子效应。
方法：我们通过将睾丸从阴囊（34°C）移位到腹部区域（37°C）并在1天后处死动物来产生隐睾大鼠模型，3天,和5天。使用淘析离心和percoll梯度方法纯化粗线精母细胞和圆形精子细胞。我们在粗线精母细胞和圆形精子细胞中进行了转录组测序，以鉴定差异表达的piRNAs及其可能的靶标。即，TE转录物和mRNA。
结果：由于热应激，我们观察到睾丸生殖细胞中piRNA和TE转录本的显著上调。除此之外,piRNA生物发生机制和热休克蛋白(Hsp70和Hsp90家族成员)上调。mRNAs也被提出作为piRNAs的靶标；因此，我们入围了某些表达成反比的piRNA-mRNA对。我们观察到在睾丸热应激中，热休克蛋白与piRNA生物发生机制的上调齐头并进。热应激生殖细胞中piRNAs的失调，增加乒乓球活动，piRNA靶转录物的表达紊乱表明piRNA之间存在联系，mRNA,和TE成绩单。
结论：在热应激中，piRNAs,piRNA机器，热休克蛋白被激活以应对低水平的压力，随后是在由高TE活性引起的长期压力中进行挽救的方法，以允许基因突变，也许是为了生存和适应。
方法：我们通过将睾丸从阴囊（34°C）移位到腹部区域（37°C）并在1天后处死动物来产生隐睾大鼠模型，3天,和5天。使用淘析离心和percoll梯度方法纯化粗线精母细胞和圆形精子细胞。我们在粗线精母细胞和圆形精子细胞中进行了转录组测序，以鉴定差异表达的piRNAs及其可能的靶标。即，TE转录物和mRNA。
结果：由于热应激，我们观察到睾丸生殖细胞中piRNA和TE转录本的显著上调。除此之外,piRNA生物发生机制和热休克蛋白(Hsp70和Hsp90家族成员)上调。mRNAs也被提出作为piRNAs的靶标；因此，我们入围了某些表达成反比的piRNA-mRNA对。我们观察到在睾丸热应激中，热休克蛋白与piRNA生物发生机制的上调齐头并进。热应激生殖细胞中piRNAs的失调，增加乒乓球活动，piRNA靶转录物的表达紊乱表明piRNA之间存在联系，mRNA,和TE成绩单。
结论：在热应激中，piRNAs,piRNA机器，热休克蛋白被激活以应对低水平的压力，随后是在由高TE活性引起的长期压力中进行挽救的方法，以允许基因突变，也许是为了生存和适应。
