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可育和不育精子的RNA动态变化

RNA dynamics of fertile and infertile spermatozoa.

作者信息

Carreau S, Lambard S, Said L, Saad A, Galeraud-Denis I

机构信息

Biochemistry Laboratory, EA 2608-USC INRA 2006, Université de Caen, Esplanade de la paix, 14032 Caen cedex, France.

出版信息

Biochem Soc Trans. 2007 Jun;35(Pt 3):634-6. doi: 10.1042/BST0350634.

Abstract

The presence of a complex population of mRNAs in human mature spermatozoa is well documented; among them, transcripts of aromatase and ERs (oestrogen receptors) have been described but their significance is not clear. Therefore, to clarify the role of this complex population of mRNAs in human ejaculated sperm, we have isolated on discontinuous density gradients two main fractions from the same sample: high- and low-motile spermatozoa. The levels of different transcripts coding for molecules involved in nuclear condensation [Prm-1 (protamine 1) and Prm-2], capacitation [eNOS (endothelial nitric oxide synthase), nNOS (neuronal nitric oxide synthase), c-myc], motility and sperm survival (aromatase) have been assessed using semi-quantitative RT (reverse transcriptase)-PCR. The viability of sperm as well as the percentage of apoptosis were identical in high- and low-motile fractions. No significant change in the c-myc/Prm-2 ratio between the two populations of spermatozoa was observed. Conversely the amount of Prm-1 mRNA was significantly higher in low-motile than in high-motile fraction; in most of the high-motile sperm samples analysed, eNOS and nNOS transcripts were undetectable, whereas they were observed in low-motile sperm. Moreover, a partial or complete disappearance of c-myc transcripts was observed after capacitation. As to the aromatase expression, a significant decrease in the amount of transcripts in immotile sperm fraction was recorded in all samples studied. To conclude, analysing mRNA profiles in humans could be helpful either as a diagnostic tool to evaluate male fertility, since they reflect spermatogenesis gene expression, and/or a prognosis value for fertilization, since these RNAs are delivered to oocytes.

摘要

人类成熟精子中存在复杂的mRNA群体,这一点已有充分记录;其中,芳香化酶和雌激素受体(ERs)的转录本已被描述,但其意义尚不清楚。因此,为了阐明这种复杂的mRNA群体在人类射出精子中的作用,我们在不连续密度梯度上从同一样本中分离出两个主要部分:高活力和低活力精子。使用半定量逆转录聚合酶链反应(RT-PCR)评估了编码参与核浓缩的分子[鱼精蛋白1(Prm-1)和鱼精蛋白2(Prm-2)]、获能[内皮型一氧化氮合酶(eNOS)、神经元型一氧化氮合酶(nNOS)、c-myc]、活力和精子存活(芳香化酶)的不同转录本的水平。高活力和低活力部分的精子活力以及凋亡百分比相同。在这两种精子群体之间未观察到c-myc/Prm-2比值有显著变化。相反,低活力部分中Prm-1 mRNA的量显著高于高活力部分;在分析的大多数高活力精子样本中,未检测到eNOS和nNOS转录本,而在低活力精子中观察到了它们。此外,在获能后观察到c-myc转录本部分或完全消失。至于芳香化酶的表达,在所有研究样本中,不动精子部分的转录本量均显著下降。总之,分析人类的mRNA谱作为评估男性生育能力的诊断工具可能会有所帮助,因为它们反映了精子发生过程中的基因表达,和/或作为受精的预后指标,因为这些RNA会传递给卵母细胞。

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