乳酸调控小鼠支持细胞增殖和基因表达的潜在机制分析

doi:10.16829/j.slxb.150778

兽类学报 ›› 2023, Vol. 43 ›› Issue (5): 568-579.DOI: 10.16829/j.slxb.150778

乳酸调控小鼠支持细胞增殖和基因表达的潜在机制分析

张晓娜^1,2,3, 贾功雪^1,2,3, 伍仕鑫^1,2,3, 万瑞东^1,2,3, 王玉军^1,2,3, 杨其恩^1,2,3

1 中国科学院西北高原生物研究所, 中国科学院高原生物适应与进化重点实验室, 西宁 810001;
2 中国科学院大学, 北京 100049;
3 中国科学院西北高原生物研究所, 青海省动物生态基因组学重点实验室, 西宁 810001

收稿日期:2023-02-14 修回日期:2023-07-20 出版日期:2023-09-30 发布日期:2023-09-22
通讯作者: 杨其恩,E-mail:yangqien@nwipb.cas.cn
作者简介:张晓娜(1991-)，女，博士研究生，主要从事动物生殖与发育研究.
基金资助:
国家自然科学基金(81960287)；中国科学院青年创新促进会(2021432)；青海省重大专项“三江源区代表性动物基因资源保护与应用”

Dissecting potential mechanisms of lactate-dependent Sertoli cell proliferation and gene expression

ZHANG Xiaona^1,2,3, JIA Gongxue^1,2,3, WU Shixin^1,2,3, WAN Ruidong^1,2,3, WANG Yujun^1,2,3, YANG Qien^1,2,3

1 Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China;
2 University of Chinese Academy of Sciences, Beijing 100049, China;
3 Qinghai Key Laboratory of Animal Ecological Genomics, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China

Received:2023-02-14 Revised:2023-07-20 Online:2023-09-30 Published:2023-09-22

摘要/Abstract

摘要： 精子发生是哺乳动物产生功能配子的细胞分化过程。作为曲细精管中唯一的体细胞，支持细胞为各级生精细胞提供结构、营养和调节支持，在精子发生中发挥不可替代的作用。睾丸中支持细胞总量是决定精子数量和生育能力的关键因素。支持细胞在胚胎发育后期和新生期睾丸中增殖，在成年睾丸中停止增殖。已发表的数据表明，支持细胞分泌的乳酸为精子发育提供能量和调控信号，但乳酸是否调控支持细胞增殖并不清楚。本研究利用免疫组化染色证实乳酸脱氢酶A (LDHA)在支持细胞中表达。利用条件性敲除小鼠模型发现，LDHA功能缺失导致支持细胞明显减少。进一步分析发现，敲除小鼠睾丸中支持细胞增殖指数在胚胎期16.5 d和出生后0 d分别下降了17.0%和9.0%，细胞凋亡的比例分别增加了86.9%和459.0%。分离纯化的支持细胞在添加10 mmol/L乳酸的培养液中培养24 h后，与对照相比，936个基因显著差异表达，其中695个表达上调，241个表达下调。GO和KEGG富集分析显示，乳酸处理支持细胞的MAPK信号通路、PI3K-AKT信号通路显著上调，而DNA复制和氧化磷酸化显著下调。根据这些研究结果，可以得出结论：LDHA在小鼠支持细胞中表达，并对调节支持细胞增殖有重要作用。

关键词: 乳酸, 支持细胞, 增殖, 凋亡

Abstract: Sertoli cells proliferate in fetal and neonatal testes and remain quiescent in adult life. Functional evidence suggests that the size of the Sertoli cell population determines sperm production and fertility. However, the factors that regulate Sertoli cell proliferation and maturation are not fully understood. Lactate, which is secreted by Sertoli cells via lactate dehydrogenase, plays a critical role in cell fate determination. We found that Ldha mRNA is present in mouse testes and immunohistochemical staining confirmed that LDHA is enriched in Sertoli cells. To study the function of LDHA in Sertoli cells, we prepared a conditional knockout mouse model using Amh-Cre and Ldha^fl/fl mice. The results showed that LDHA deficiency resulted in a significant reduction in the number of Sertoli cells, which was (18. 27 ±0. 60) /cell in Ldha-cKO mice compared to (21. 1 ±0. 68) /cell in control mice. Further analysis revealed that the proliferative capacity of Sertoli cells in Ldha-cKO mice decreased by 17. 0% and 9. 0% at Embryonic day 16. 5 and Day 0, respectively, while cell apoptosis increased by 86. 9% and 459. 0%, respectively. After incubation with 10 mmol/L lactic acid for 24 h, 936 significantly differentially expressed genes were identified in Sertoli cells of the knockout mouse, of which 695 were upregulated and 241 were downregulated. GO and KEGG enrichment analyses revealed that the MAPK pathway and PI3K-AKT signaling pathway were upregulated, while DNA replication and oxidative phosphorylation were downregulated. Hence, it can be concluded that LDHA is expressed in mouse Sertoli cells and plays an important role in regulating Sertoli cell proliferation.

Key words: Lactate, Sertoli cells, Proliferation, Apoptosis

中图分类号:

Q492

张晓娜, 贾功雪, 伍仕鑫, 万瑞东, 王玉军, 杨其恩. 乳酸调控小鼠支持细胞增殖和基因表达的潜在机制分析[J]. 兽类学报, 2023, 43(5): 568-579.

ZHANG Xiaona, JIA Gongxue, WU Shixin, WAN Ruidong, WANG Yujun, YANG Qien. Dissecting potential mechanisms of lactate-dependent Sertoli cell proliferation and gene expression[J]. ACTA THERIOLOGICA SINICA, 2023, 43(5): 568-579.

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乳酸调控小鼠支持细胞增殖和基因表达的潜在机制分析

Dissecting potential mechanisms of lactate-dependent Sertoli cell proliferation and gene expression

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