Glis3 plays a crucial role in spermatogenesis [848_827]

We show that Glis3 is expressed in gonocytes, SSCs and SPCs, but not in differentiated spermatogonia or subsequent stages of spermatogenesis nor in Sertoli or Leydig cells. We further demonstrate that Glis3-deficiency causes a severe impairment in spermatogenesis in mice. Although the number of gonocytes was slightly diminished in Glis3KO testis, the number undifferentiated, PLZF+ spermatogonia was dramatically reduced leading to a virtual block in the progression of spermatogenesis. Gene expression profiling showed that the expression of a number of genes associated with self-renewal and differentiation of spermatogonial cells was significantly decreased in 1-week-old Glis3KO2 testis. These included a set of GDNF-dependent genes, such as Etv5, Bcl6b, Lhx1, Brachyury, Id4, and Pou3f1, and GDNF-independent genes, such as FoxO1, Oct4, and Zbtb16. Impairment of the nuclear localization of FoxO1 may be in part responsible for the reduced expression of Ret, Lhx1, and Sall4 in Glis3KO2 testis. Our study identifies Glis3 as a novel and critical regulator of early stages of spermatogenesis. Testis total RNAs were purified from 4 WT and 4 Glis3KO2 at 1 week old age, and 3WT and 3 Glis3KO2 at 3 week-old age. Then the samples were applied to Agilent mouse genome chip.本研究证实，Glis3可在生殖母细胞（gonocytes）、精原干细胞（Spermatogonial Stem Cells, SSCs）以及精原祖细胞（Spermatogonial Progenitor Cells, SPCs）中表达，但在分化型精原细胞、精子发生后续阶段，以及支持细胞（Sertoli cells）与间质细胞（Leydig cells）中均无表达。本研究进一步证实，Glis3基因缺失会导致小鼠精子发生出现严重障碍。尽管Glis3基因敲除（Glis3KO）小鼠睾丸内的生殖母细胞数量仅略有减少，但未分化的早幼粒细胞白血病锌指蛋白阳性（PLZF+）精原细胞数量却显著降低，进而造成精子发生进程几乎完全阻滞。基因表达谱分析显示，在1周龄Glis3KO2小鼠睾丸中，一系列与精原细胞自我更新及分化相关的基因表达水平显著下调。其中既包含胶质细胞源性神经营养因子（GDNF）依赖型基因，如Etv5、Bcl6b、Lhx1、Brachyury、Id4及Pou3f1，也涵盖GDNF非依赖型基因，如FoxO1、Oct4及Zbtb16。FoxO1的核定位功能受损，可能是Glis3KO2小鼠睾丸中Ret、Lhx1及Sall4基因表达下调的部分原因。本研究鉴定出Glis3是精子发生早期阶段的新型关键调控因子。实验中，我们从1周龄的4只野生型（Wild Type, WT）小鼠与4只Glis3KO2小鼠，以及3周龄的3只WT小鼠与3只Glis3KO2小鼠体内提取睾丸总RNA，随后将所有样本应用于安捷伦（Agilent）小鼠全基因组芯片进行检测。