Tight gene co-expression in BCB positive cattle oocytes and their surrounding cumulus cells.

Bailey N Walker, Jada Nix, Chace Wilson, Mackenzie A Marrella, Savannah L Speckhart, Lydia Wooldridge, Con-Ning Yen, Jocelyn S Bodmer, Laila T Kirkpatrick, Sarah E Moorey, David E Gerrard, Alan D Ealy, Fernando H Biase
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引用次数: 5

Abstract

Background: Cytoplasmic and nuclear maturation of oocytes, as well as interaction with the surrounding cumulus cells, are important features relevant to the acquisition of developmental competence.

Methods: Here, we utilized Brilliant cresyl blue (BCB) to distinguish cattle oocytes with low activity of the enzyme Glucose-6-Phosphate Dehydrogenase, and thus separated fully grown (BCB positive) oocytes from those in the growing phase (BCB negative). We then analyzed the developmental potential of these oocytes, mitochondrial DNA (mtDNA) copy number in single oocytes, and investigated the transcriptome of single oocytes and their surrounding cumulus cells of BCB positive versus BCB negative oocytes.

Results: The BCB positive oocytes were twice as likely to produce a blastocyst in vitro compared to BCB- oocytes (P < 0.01). We determined that BCB negative oocytes have 1.3-fold more mtDNA copies than BCB positive oocytes (P = 0.004). There was no differential transcript abundance of genes expressed in oocytes, however, 172 genes were identified in cumulus cells with differential transcript abundance (FDR < 0.05) based on the BCB staining of their oocyte. Co-expression analysis between oocytes and their surrounding cumulus cells revealed a subset of genes whose co-expression in BCB positive oocytes (n = 75) and their surrounding cumulus cells (n = 108) compose a unique profile of the cumulus-oocyte complex.

Conclusions: If oocytes transition from BCB negative to BCB positive, there is a greater likelihood of producing a blastocyst, and a reduction of mtDNA copies, but there is no systematic variation of transcript abundance. Cumulus cells present changes in transcript abundance, which reflects in a dynamic co-expression between the oocyte and cumulus cells.

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紧密基因在BCB阳性牛卵母细胞及其周围积云细胞中的共表达。
背景:卵母细胞的细胞质和核成熟以及与周围积云细胞的相互作用是获得发育能力的重要特征。方法:利用亮甲酰蓝(BCB)对葡萄糖-6-磷酸脱氢酶活性较低的牛卵母细胞进行区分,将发育完全的(BCB阳性)卵母细胞与处于生长期的(BCB阴性)卵母细胞分离。然后,我们分析了这些卵母细胞的发育潜力,单个卵母细胞的线粒体DNA (mtDNA)拷贝数,并研究了BCB阳性卵母细胞和BCB阴性卵母细胞的单个卵母细胞及其周围卵丘细胞的转录组。结果:BCB阳性卵母细胞体外产生囊胚的可能性是BCB阳性卵母细胞的两倍(P结论:当卵母细胞从BCB阴性转变为BCB阳性时,产生囊胚的可能性更大,mtDNA拷贝数减少,但转录物丰度没有系统性变化。积云细胞转录物丰度的变化反映在卵母细胞和积云细胞之间的动态共表达。
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