Expression of PCOS candidate genes in bovine fetal and adult ovarian somatic cells.

Menghe Liu, Nicole A Bastian, Monica Dwi Hartanti, Katja Hummitzsch, Helen F Irving-Rodgers, Richard A Anderson, Raymond Joseph Rodgers
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Abstract

Polycystic ovary syndrome (PCOS) is an endocrine metabolic disorder that appears to have a genetic predisposition and a fetal origin. The fetal ovary has two major somatic cell types shown previously to be of different cellular origins, different morphologies and to differentially express 15 genes. We isolated the somatic gonadal ridge epithelial-like (GREL) cells (n = 7) and ovarian fetal fibroblasts (n = 6) by clonal expansion. Using qRT-PCR, we compared the gene expression levels of PCOS candidate genes with previous data on the expression levels in whole fetal ovaries across gestation. We also compared these levels with those in bovine adult ovarian cells including fibroblasts (n = 4), granulosa cells (n = 5) and surface epithelial cells (n = 5). Adult cell types exhibited clear differences in the expression of most genes. In fetal ovarian cells, DENND1A and ERBB3 had significantly higher expression in GREL cells. HMGA2 and TGFB1I1 tended to have higher expression in fetal fibroblasts than GREL cells. Another 19 genes did not exhibit differences between GREL cells and fetal fibroblasts and FBN3, FSHB, LHCGR, FSHR and ZBTB16 were very lowly expressed in GREL cells and fibroblasts. The culture of fetal fibroblasts in EGF-containing medium resulted in lower expression of NEIL2, but higher expression of MAPRE1 compared to culture in the absence of EGF. Thus, the two fetal ovarian somatic cell types mostly lacked differential expression of PCOS candidate genes.

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牛胎儿和成年卵巢体细胞中 PCOS 候选基因的表达。
多囊卵巢综合征(PCOS)是一种内分泌代谢紊乱疾病,似乎具有遗传倾向,并且起源于胎儿。以前的研究表明,胎儿卵巢有两种主要的体细胞类型,它们具有不同的细胞起源、不同的形态以及 15 个基因的不同表达。我们通过克隆扩增分离了体细胞性腺脊上皮样(GREL)细胞(n = 7)和卵巢胎儿成纤维细胞(n = 6)。通过 qRT-PCR,我们将多囊卵巢综合征候选基因的基因表达水平与之前关于整个妊娠期胎儿卵巢表达水平的数据进行了比较。我们还比较了这些基因在牛成年卵巢细胞中的表达水平,包括成纤维细胞(4 个)、颗粒细胞(5 个)和表面上皮细胞(5 个)。成体细胞类型在大多数基因的表达上表现出明显的差异。在胎儿卵巢细胞中,DENND1A 和 ERBB3 在 GREL 细胞中的表达量明显较高。HMGA2和TGFB1I1在胎儿成纤维细胞中的表达量往往高于GREL细胞。另外 19 个基因在 GREL 细胞和胎儿成纤维细胞中没有表现出差异,FBN3、FSHB、LHCGR、FSHR 和 ZBTB16 在 GREL 细胞和成纤维细胞中的表达量很低。与不含 EGF 的培养基相比,在含 EGF 的培养基中培养胎儿成纤维细胞,NEIL2 的表达较低,但 MAPRE1 的表达较高。因此,两种胎儿卵巢体细胞类型大多缺乏 PCOS 候选基因的差异表达。
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