Pyruvate dehydrogenase complex: mRNA and protein expression patterns of E1¦Á subunit genes in human spermatogenesis

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• 作者：Ana Pinheiro ; Maria Jo?o Silva ; In¨ºs Gra?a ; Joaquina Silva ; Ros¨¢lia S¨¢ ; M¨¢rio Sousa ; Alberto Barros ; Isabel Tavares de Almeida ; Isabel Rivera
• 关键词：AZF ; azoospermia factor ; IEF ; isoelectric focusing ; IP ; isoelectric point ; IPF ; immobilized pH gradient ; OSCP ; oligomycin sensitivity conferral protein (subunit of ATP synthase) ; PDC ; pyruvate dehydrogenase complex ; PDHA1 ; somatic isoform of pyruvate dehy
• 刊名：Gene
• 出版年：2012
• 出版时间：10 September, 2012
• 年：2012
• 卷：506
• 期：1
• 页码：173-178
• 全文大小：1095 K

文摘

During spermatogenesis, germ cells undergo a complex process of cell differentiation and morphological restructuring, which depends on the coordinated expression of different genes. Some vital examples are those involved in cell energy metabolism, namely the genes encoding the E1¦Á subunit of pyruvate dehydrogenase complex: the somatic PDHA1 (X-linked) and the testis-specific PDHA2 (autosomal). There are no data related to the study at the RNA and protein levels of PDHA genes during human spermatogenesis. The present study aimed to describe the mRNA and protein expression patterns of the human PDHA genes during spermatogenesis.

Expression profiles of the PDHA1 and PDHA2 genes were characterized using different human tissues and cells. Diploid and haploid germ cells fractions were obtained from testis tissues. The mRNA profiles were analyzed by quantitative RT-PCR, whereas the protein profiles were evaluated by immunohistochemistry, western blotting and two-dimensional electrophoresis.

Expression of the PDHA1 gene was found in all somatic cells, whereas expression of PDHA2 gene was restricted to germ cells. The switch from X-linked to autosomic gene expression occurred in spermatocytes. Data suggest the activation of PDHA2 gene expression is most probably a mechanism to ensure the continued expression of the protein, thus allowing germ cell viability and functionality.