背景:探索原始生殖细胞(PGC)迁移和性腺体细胞参与性腺发育的分子机制对于理解生殖相关疾病的起源和潜在治疗具有重要意义。
方法:通过分析公开可用的数据集(ATAC-seq,DNase-seq,和RNA-seq)。随后,采用CRISPR-Cas9技术构建Diaph1基因敲除小鼠,研究Diaph1在性腺发育中的作用。
结果:基于来自公共数据库的数据,在小鼠PGC的迁移中鉴定出差异表达基因Diaph1。此外,与野生型小鼠相比,Diaph1基因敲除小鼠的PGCs数量显著减少,和增殖相关基因(Dicer1,Mcm9)的表达水平,粘附(E-cadherin,Cdh1),和迁移(Cxcr4、Hmgcr、Dazl)显著降低。Diaph1基因敲除还能抑制睾丸间质细胞增殖并诱导其凋亡,以及卵巢颗粒细胞凋亡。此外,Diaph1基因敲除小鼠的附睾区精子数和卵巢卵泡数明显减少,导致生育率下降,伴随着血清睾酮和雌二醇水平的降低。进一步研究发现,在Diaph1基因敲除小鼠中,睾丸间质细胞中睾酮合成的关键酶(CYP11A1,3β-HSD)减少,和雌二醇相关因子(FSH受体,颗粒细胞中的AMH)也下调。
结论:总体而言,我们的发现表明敲除Diaph1可以破坏调节性激素产生的因子的表达,导致性激素分泌受损,最终导致生殖功能受损。这些结果为PGC迁移和性腺发育的分子机制提供了新的视角,并为进一步研究原因提供有价值的见解,诊断,以及相关疾病的治疗。
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.