Analysis of cell-type-specific gene expression during mouse spermatogenesis
Analysis of cell-type-specific gene expression during mouse spermatogenesis
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DOI：
10.1095/biolreprod.103.026575
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发表时间：
2004-06-01
影响因子：
3.6
通讯作者：
Leffers, H
中科院分区：
文献类型：
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作者：
Almstrup, K;Nielsen, JE;Leffers, H
在啮齿类动物中，精子发生过程中基因表达的变化可以通过产后发育过程中每天的睾丸取样来监测。然而，组织水平基因表达的变化可以反映特定细胞类型中mRNA浓度的变化、特定细胞体积的变化或细胞类型组成的变化。这反映了组织的细胞结构。在这里，我们结合了在全组织水平、差异显示和 DNA 阵列以及细胞水平、原位杂交评估基因表达谱的技术。结合这些技术的结果可以确定精子发生过程中许多基因的细胞类型特异性基因表达模式。差异显示用于以高灵敏度确定表达谱，并且与序列的先验知识无关，而DNA阵列可以快速评估所有基因的表达谱。这确定了三组基因表达谱。主要组对应于在精子发生的中粗线期或粗线期晚期（VII-XI 阶段）精母细胞中上调的基因。该粗线期簇在精子细胞的后期阶段逐渐消失，但随后在精子细胞中表达了另一簇基因。最后，一组基因在精子发生过程中被下调，并可能在非生殖细胞中表达。我们相信大多数基因的表达可以通过这些细胞类型特异性表达模式的组合来描述。
In rodents, changes in gene expression during spermatogenesis can be monitored by sampling testis from each day during postnatal development. However, changes in gene expression at the tissue level can reflect changes in the concentration of an mRNA in a specific cell type, changes in volume of specific cells, or changes in the cell-type composition. This reflects the cellularity of the tissue. Here we have combined techniques that assess the expression profiles of genes at the whole-tissue level, differential display and DNA array, and, at the level of cellularity, in situ hybridization. Combining results from these techniques allows determination of the cell-type-specific gene-expression patterns of many genes during spermatogenesis. Differential display was used to determine expression profiles with high sensitivity and independent of prior knowledge of the sequence, whereas DNA arrays quickly assess the expression profiles of all the genes. This identified three groups of gene-expression profiles. The major group corresponds to genes that are upregulated in spermatocytes during either the mid- or latepachytene phase of spermatogenesis (stages VII-XI). This pachytene cluster was gradually extinguished in the later spermatid stages but was followed by another cluster of genes expressed in spermatids. Finally, a group of genes was downregulated during spermatogenesis and probably expressed in nongerm cells. We believe that expression of most genes can be described by a combination of these cell-type-specific expression patterns.