Brevini-Gandolfi T A, Favetta L A, Mauri L, Luciano A M, Cillo F, Gandolfi F
Department of Endocrinology, University of Milan, Italy.
Mol Reprod Dev. 1999 Apr;52(4):427-33. doi: 10.1002/(SICI)1098-2795(199904)52:4<427::AID-MRD12>3.0.CO;2-G.
Molecules of mRNA are stored in the oocyte cytoplasm in order to be used during the initial phases of embryonic development. The storage takes place during oocyte growth and the extent of poly(A) tail at the 3' end of the transcripts has emerged as an important regulatory element for determining their stability. The objective of the present study was to analyse changes in polyadenylation levels of mRNA transcripts, stored in bovine oocytes, during in vitro maturation and their possible relation with developmental competence. Oocyte developmental competence was predicted on the basis of the morphological appearance of their originating ovary as previously established (Gandolfi et al. 1997a. Theriogenology 48:1153-1160) and were divided into groups H (high competence) and L (low competence). The length of the poly(A) tail of the following genes, beta-actin (beta-Act), connexin 43, glucose transporter type 1, heat shock protein 70, oct-4, plakophilin, pyruvate dehydrogenase phosphatase (PDP), and RNA poly(A) polymerase, was determined at the germinal vesicle (GV) and metaphase II (MII) stage. The results indicated that the poly(A) tail of all genes except for beta-Act and PDP, is shorter after in vitro maturation (IVM) in both groups. Moreover, group L oocytes showed a shorter poly(A) tail than group H oocytes in all genes except for beta-Act and PDP, both at GV and MII stage. We conclude that most of the examined transcripts follow the default deadenylation pattern described during oocyte maturation in other species and that a shorter poly(A) tail is correlated with low developmental competence.
mRNA分子存储在卵母细胞的细胞质中,以便在胚胎发育的初始阶段使用。这种存储发生在卵母细胞生长期间,转录本3'端的聚腺苷酸(poly(A))尾长度已成为决定其稳定性的重要调控元件。本研究的目的是分析体外成熟过程中存储在牛卵母细胞中的mRNA转录本的多聚腺苷酸化水平变化及其与发育能力的可能关系。根据先前确定的(Gandolfi等人,1997a。《兽医学》48:1153 - 1160)其来源卵巢的形态外观预测卵母细胞的发育能力,并将其分为H组(高能力)和L组(低能力)。在生发泡(GV)期和中期II(MII)期测定以下基因β-肌动蛋白(β-Act)、连接蛋白43、葡萄糖转运蛋白1型、热休克蛋白70、八聚体结合转录因子4、桥粒斑蛋白、丙酮酸脱氢酶磷酸酶(PDP)和RNA聚腺苷酸聚合酶的poly(A)尾长度。结果表明,除β-Act和PDP外,两组中所有基因的poly(A)尾在体外成熟(IVM)后均变短。此外,在GV期和MII期,除β-Act和PDP外,L组卵母细胞在所有基因中的poly(A)尾均比H组卵母细胞短。我们得出结论,大多数检测的转录本遵循其他物种卵母细胞成熟过程中描述的默认去腺苷酸化模式,且较短的poly(A)尾与低发育能力相关。