Biopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City University, Yokohama, Kanagawa, Japan; Laboratory for Cellular Function Conversion Technology, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa, Japan.
Laboratory for Cellular Function Conversion Technology, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa, Japan.
Biochem Biophys Res Commun. 2020 Oct 1;530(4):732-738. doi: 10.1016/j.bbrc.2020.06.161. Epub 2020 Aug 8.
Cultivation of neonatal mouse testis tissue can induce spermatogenesis and produce fertile sperms. However, in vitro spermatogenesis mediated by the current organ culture method comes short in fully mimicking the in vivo counterpart, partly due to a lack of knowledge underlying molecular phenotypes of in vitro spermatogenesis. In this study, we investigated transcriptome of cultured testis tissues using microarray method. Principle component analysis of the transcriptome data revealed delay and/or arrest of spermatogenesis and immediate radical immune reactions in the cultured testis tissues. The delay/arrest of spermatogenesis occurred before and during early meiotic phase, resulting in inefficient progression of meiosis. The immune reaction, on the other hand, was drastic and overwhelming, in which TLR4-NF-kB signaling was speculated to be involved. Notably, treatment with TAK242, an inhibitor of TLR4-NF-kB signaling pathway, ameliorated the macrophage activation which otherwise would exacerbate the inflammation. Thus, the present study revealed for the first time at molecular level that the deficiency of germ cell differentiation and the immense immune reaction are major abnormalities in the cultured testis tissues.
培养新生小鼠睾丸组织可以诱导精子发生并产生可育精子。然而，目前的器官培养方法介导的体外精子发生在很大程度上不能完全模拟体内对应物，部分原因是缺乏对体外精子发生分子表型的了解。在这项研究中，我们使用微阵列方法研究了培养的睾丸组织的转录组。转录组数据的主成分分析表明，培养的睾丸组织中存在精子发生延迟和/或停滞以及即刻的激进免疫反应。精子发生的延迟/停滞发生在减数分裂早期之前和期间，导致减数分裂效率低下。另一方面，免疫反应是剧烈和压倒性的，推测其中涉及 TLR4-NF-κB 信号通路。值得注意的是，TLR4-NF-κB 信号通路抑制剂 TAK242 的治疗改善了巨噬细胞的激活，否则会加剧炎症。因此，本研究首次从分子水平揭示了生殖细胞分化缺陷和巨大免疫反应是培养的睾丸组织中的主要异常。
