Comparative study on AZF gene expression in peripheral leukocytes and testicular cells on idiopathic infertility males

Abstract

Objective To investigate the difference on AZF gene expression between peripheral leukocytes and testicular cells of idiopathic infertility males.Methods Sixty-two idiopathic infertility patients were enrolled.The infertility group included 33 males with azoospermia and 29 with severe oligozoospermia.Genomic DNA was prepared from peripheral blood samples of the 62 patients.Each patient was tested for STS of AZF region located at long arm of Y-chromosome,including sY84 and sY86 for AZFa,sY127 and sY134 for AZFb,sY254 and sY255 for AZFc.The two multiplex PCR consisted of:MixA:SRY(sY14)-ZFY-sY84-sY134-sY255 and MixB:SRY(sY14)-ZFY-sY86-sY127-sY254.The testicular material of infertile men was obtained by bilateral fine needle aspiration cytology.Each patient was tested for STS of AZF region located at long arm of Y-chromosome,including DFFRY for AZFa,RBM for AZFb,sY254(DAZ)for AZFc.The PCR consisted of:SRY(sYl4)-DFFRY-RBM-DAZ(sY254)-β-actin.Results In 62 cases,12 deletions(19.4%)wet found.Nine patients(9/33,27.3%)RT-PCR analysis from testicular cells gave normal amplifications for SRY and DFFRY mRNA in 62 idiopathic patients.Two patients were negative for RBM expression.No RBM and DAZ were detected in 1 case.Twelve patients had no expression in the AZFc region involving the DAZ gene.Of the 12 cases,3 patients with normal PCR analysis of DAZ gene on genomic DNA.Conclusions AZF microdeletion detection is possible in testicular material.Hence RT-PCR analysis can also be helpful to define etiology of idiopathic infertility. Key words: Y chromosome; A zoospermia factor; Polymerase chain reaction; Reverse transcriptase polymerase chain reaction; Infertility,male