A bioenergetic balance between glycolysis and mitochondrial respiration is particularly important for stem cell fate specification. It however remains to be determined whether undifferentiated spermatogonia switch their preference for bioenergy production during differentiation. In this study, we found that ATP generation in spermatogonia was gradually increased upon retinoic acid (RA)-induced differentiation. To accommodate this elevated energy demand, RA signaling concomitantly switched ATP production in spermatogonia from glycolysis to mitochondrial respiration, accompanied by increased levels of reactive oxygen species. Disrupting mitochondrial respiration significantly blocked spermatogonial differentiation. Inhibition of glucose conversion to glucose-6-phosphate or pentose phosphate pathway also repressed the formation of c-Kit⁺ differentiating germ cells, suggesting that metabolites produced from glycolysis are required for spermatogonial differentiation. We further demonstrated that the expression levels of several metabolic regulators and enzymes were significantly altered upon RA-induced differentiation, with both RNA-seq and quantitative proteomic analyses. Taken together, our data unveil a critically regulated bioenergetic balance between glycolysis and mitochondrial respiration that is required for spermatogonial proliferation and differentiation.

糖酵解和线粒体呼吸之间的生物能平衡对于干细胞命运规范尤其重要。然而，未分化的精原细胞在分化过程中是否改变了对生物能生产的偏好仍有待确定。在这项研究中，我们发现精原细胞中 ATP 的生成在视黄酸 (RA) 诱导分化后逐渐增加。为了适应这种增加的能量需求，RA 信号传导同时将精原细胞中 ATP 的产生从糖酵解转变为线粒体呼吸，同时活性氧水平增加。扰乱线粒体呼吸显着阻碍精原细胞分化。^{抑制葡萄糖转化为葡萄糖-6-磷酸或磷酸戊糖途径也抑制了c-Kit +}分化生殖细胞的形成，这表明糖酵解产生的代谢物是精原细胞分化所必需的。我们通过 RNA-seq 和定量蛋白质组学分析进一步证明，在 RA 诱导的分化过程中，几种代谢调节因子和酶的表达水平发生了显着改变。综上所述，我们的数据揭示了精原细胞增殖和分化所需的糖酵解和线粒体呼吸之间严格调节的生物能量平衡。