Gilbert Isabelle, Bissonnette Nathalie, Boissonneault Guylain, Vallée Maud, Robert Claude
Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Université Laval, Québec G1K 7P4, Canada.
Reproduction. 2007 Jun;133(6):1073-86. doi: 10.1530/REP-06-0292.
Spermiogenesis represents the transition from haploid spermatids to spermatozoa. This process entails an extreme condensation of the nucleus and a loss of nearly all cytoplasmic content. The presence of messenger RNAs in the spermatozoa has previously been shown. Generally, these transcripts are considered to be remnants of spermiogenesis. However, it has recently been proposed that there may exist a function for these sperm-associated RNAs. To address the possibility of a functional role for these transcripts, we sought to investigate and characterize the RNA pool found in bovine spermatozoa. The main goals of this study were to examine RNA integrity and survey the mRNA found in spermatids and spermatozoa. Assessment of mRNAs integrity was performed by three approaches: microelectrophoresis, comparative smearing after global amplification, and PCR amplification of target sequences located either in the 5' or the 3' ends, while mRNAs survey was performed by microarray hybridizations. RNA integrity studies in the spermatozoa showed a majority of low molecular size fragments indicating a natural segmentation of the mRNA population. The mRNA survey indicated that the sperm transcriptome harbors a complex mixture of messengers implicated in a wide array of cell functions and representing a large subset of transcripts found in spermatids. Subsequently, such sperm RNA profiling could allow the molecular diagnosis of male gamete quality.
精子形成代表了从单倍体精子细胞到精子的转变。这个过程需要细胞核的极度浓缩以及几乎所有细胞质内容物的丢失。先前已经证明精子中存在信使核糖核酸。一般来说，这些转录本被认为是精子形成的残余物。然而，最近有人提出这些与精子相关的核糖核酸可能存在某种功能。为了探究这些转录本是否具有功能作用，我们试图研究并表征牛精子中发现的核糖核酸库。本研究的主要目标是检查核糖核酸的完整性，并调查精子细胞和精子中发现的信使核糖核酸。通过三种方法评估信使核糖核酸的完整性：微电泳、全局扩增后的比较涂片以及对位于5'或3'端的靶序列进行聚合酶链反应扩增，而通过微阵列杂交进行信使核糖核酸调查。精子中的核糖核酸完整性研究表明，大多数是低分子大小的片段，这表明信使核糖核酸群体存在自然分割。信使核糖核酸调查表明，精子转录组包含复杂的信使核糖核酸混合物，这些信使核糖核酸涉及广泛的细胞功能，并且代表了精子细胞中发现的大量转录本子集。随后，这种精子核糖核酸谱分析可以实现对雄配子质量的分子诊断。
