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Abstract
BackgroundWe generated the gene expression profile of the total testis from the adult C57BL/6J male mice using serial analysis of gene expression (SAGE). Two high-quality SAGE libraries containing a total of 76 854 tags were constructed. An extensive bioinformatic analysis and comparison of SAGE transcriptomes of the total testis, testicular somatic cells and other mouse tissues was performed and the theory of male-biased gene accumulation on the X chromosome was tested.ResultsWe sorted out 829 genes predominantly expressed from the germinal part and 944 genes from the somatic part of the testis. The genes preferentially and specifically expressed in total testis and testicular somatic cells were identified by comparing the testis SAGE transcriptomes to the available transcriptomes of seven non-testis tissues. We uncovered chromosomal clusters of adjacent genes with preferential expression in total testis and testicular somatic cells by a genome-wide search and found that the clusters encompassed a significantly higher number of genes than expected by chance. We observed a significant 3.2-fold enrichment of the proportion of X-linked genes specific for testicular somatic cells, while the proportions of X-linked genes specific for total testis and for other tissues were comparable. In contrast to the tissue-specific genes, an under-representation of X-linked genes in the total testis transcriptome but not in the transcriptomes of testicular somatic cells and other tissues was detected.ConclusionOur results provide new evidence in favor of the theory of male-biased genes accumulation on the X chromosome in testicular somatic cells and indicate the opposite action of the meiotic X-inactivation in testicular germ cells.
Highlights
We generated the gene expression profile of the total testis from the adult C57BL/ 6J male mice using serial analysis of gene expression (SAGE)
Both total testis SAGE libraries provided similar gene expression profiles (R2 = 0.84 for all unique tags, Pearson correlation), which suggests a good reproducibility of SAGE data
A certain variation was observed in the tag abundances when 24 529 unique tags found in both total testis SAGE libraries were compared by Monte Carlo simulations
Summary
We generated the gene expression profile of the total testis from the adult C57BL/ 6J male mice using serial analysis of gene expression (SAGE). * excluding tags from duplicated ditags and linker-derived tags (i.e. two linker tags TCCCTATTAA, TCCCCGTACA and all possible 1-bp linker tag variations) modifications of paternal and maternal DNA such as reactivation of the X chromosome in female germ cells [3,4], inactivation of a single X chromosome in pachytene spermatocytes [5,6,7] or differential establishment of imprinting marks on paternally or maternally imprinted genes [8,9]. Obeying the Haldane's rule, hybrid sterility preferentially affects gametogenesis in testis in species with heterogametic (XY) sex [13,14,15]. Molecular analyses of these phenomena are hindered by the fact that testis is a complex organ with many types of intimately intermingled somatic and germline cells. The main cell types can be fractionated, via gravity sedimentation, centrifugal elutriation or fluorescence activated cell sorting, but the time required can be fairly long to exclude possible artificial changes of mRNA levels
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