Diaph1 敲除抑制小鼠原始生殖细胞增殖并影响性腺发育。

Diaph1 knockout inhibits mouse primordial germ cell proliferation and affects gonadal development.

机构信息

State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, 010020, Inner Mongolia, China.

School of Biology and Agriculture, Shaoguan University, Shaoguan, 512005, Guangdong Province, China.

出版信息

Reprod Biol Endocrinol. 2024 Jul 15;22(1):82. doi: 10.1186/s12958-024-01257-z.

DOI:10.1186/s12958-024-01257-z

PMID:39010074

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11247884/

Abstract

BACKGROUND

Exploring the molecular mechanisms of primordial germ cell (PGC) migration and the involvement of gonadal somatic cells in gonad development is valuable for comprehending the origins and potential treatments of reproductive-related diseases.

METHODS

Diaphanous related formin 1 (Diaph1, also known as mDia1) was screened by analyzing publicly available datasets (ATAC-seq, DNase-seq, and RNA-seq). Subsequently, the CRISPR-Cas9 technology was used to construct Diaph1 knockout mice to investigate the role of Diaph1 in gonad development.

RESULTS

Based on data from public databases, a differentially expressed gene Diaph1, was identified in the migration of mouse PGC. Additionally, the number of PGCs was significantly reduced in Diaph1 knockout mice compared to wild type mice, and the expression levels of genes related to proliferation (Dicer1, Mcm9), adhesion (E-cadherin, Cdh1), and migration (Cxcr4, Hmgcr, Dazl) were significantly decreased. Diaph1 knockout also inhibited Leydig cell proliferation and induced apoptosis in the testis, as well as granulosa cell apoptosis in the ovary. Moreover, the sperm count in the epididymal region and the count of ovarian follicles were significantly reduced in Diaph1 knockout mice, resulting in decreased fertility, concomitant with lowered levels of serum testosterone and estradiol. Further research found that in Diaph1 knockout mice, the key enzymes involved in testosterone synthesis (CYP11A1, 3β-HSD) were decreased in Leydig cells, and the estradiol-associated factor (FSH receptor, AMH) in granulosa cells were also downregulated.

CONCLUSIONS

Overall, our findings indicate that the knockout of Diaph1 can disrupt the expression of factors that regulate sex hormone production, leading to impaired secretion of sex hormones, ultimately resulting in damage to reproductive function. These results provide a new perspective on the molecular mechanisms underlying PGC migration and gonadal development, and offer valuable insights for further research on the causes, diagnosis, and treatment of related diseases.

摘要

背景

探索原始生殖细胞（PGC）迁移的分子机制以及性腺体细胞在性腺发育中的作用，对于理解生殖相关疾病的起源和潜在治疗方法具有重要意义。

方法

通过分析公共数据集（ATAC-seq、DNase-seq 和 RNA-seq）筛选出 Diahanas 相关formin1（Diaph1，也称为 mDia1）。随后，使用 CRISPR-Cas9 技术构建 Diaph1 敲除小鼠，以研究 Diaph1 在性腺发育中的作用。

结果

基于公共数据库中的数据，发现了一个在小鼠 PGC 迁移中差异表达的基因 Diaph1。与野生型小鼠相比，Diaph1 敲除小鼠的 PGC 数量明显减少，与增殖相关的基因（Dicer1、Mcm9）、黏附相关的基因（E-cadherin、Cdh1）和迁移相关的基因（Cxcr4、Hmgcr、Dazl）的表达水平显著降低。Diaph1 敲除还抑制了睾丸间质细胞的增殖并诱导其凋亡，以及卵巢颗粒细胞的凋亡。此外，Diaph1 敲除小鼠的附睾区域精子计数和卵巢卵泡计数明显减少，导致生育能力下降，同时血清睾酮和雌二醇水平降低。进一步的研究发现，在 Diaph1 敲除小鼠中，睾丸间质细胞中参与睾酮合成的关键酶（CYP11A1、3β-HSD）减少，颗粒细胞中与雌二醇相关的因子（FSH 受体、AMH）也下调。