Investigation of DAZ and RBMY1 gene expression in human testis by quantitative real-time PCR.
This study developed quantitative real-time PCR assays for the DAZ and RBMY1 genes to determine the copy number of RNA extracted from testicular biopsies from a cohort of normospermic controls (n=6) and azoospermic males (n=17) including two males with Y-chromosome microdeletions (AZFc and AZFb + c). All patients underwent testicular sperm extraction (TESE) for intracytoplasmic sperm injection (ICSI). Forty percent of the azoospermic cohort showed a significant reduction in the copies of at least one of the genes (DAZ P=0.003; RBMY1 P=0.009). The histopathology of these patients ranged from Sertoli cell only (SCO) to severe hypospermatogenesis with interstitial fibrosis. The patient with the AZFb + c deletion lacked expression of DAZ and RBMY1 and had a histopathology of SCO. The patient with the AZFc deletion had reduced expression of RBMY1 and no DAZ expression with a histopathology of spermatocyte arrest. The quantitative real-time PCR assays for DAZ and RBMY1 gave positive predictive values of 78% and 70%, respectively for the recovery of sperm from testicular biopsy.
Lavery
R
R
National Diagnostics Centre, National University of Ireland, Galway, Ireland.
Glennon
M
M
Houghton
J
J
Nolan
A
A
Egan
D
D
Maher
M
M
D000199:Actins / Q000235:genetics; D053713:Azoospermia / Q000235:genetics*; D001706:Biopsy; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D000075364:Deleted in Azoospermia 1 Protein; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D009687:Nuclear Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D016601:RNA-Binding Proteins / Q000235:genetics*; D012016:Reference Values; D013737:Testis / Q000166:cytology* / Q000473:pathology
D000199:Actins; C095668:DAZ1 protein, human; D017931:DNA Primers; D000075364:Deleted in Azoospermia 1 Protein; D009687:Nuclear Proteins; C501499:RBMY1A1 protein, human; D016601:RNA-Binding Proteins; D004247:DNA