title: A preliminary report on the implication of RT-PCR detection of DAZ, RBMY1, USP9Y and Protamine-2 mRNA in testicular biopsy samples from azoospermic men.
abstract: In this study, reverse transcription-polymerase chain reaction (RT-PCR) was optimized to analyse the presence of DAZ, RBMY1, USP9Y, protamine-2, SRY and actin messenger RNA (mRNA) in testicular cells of men suffering from idiopathic azoospermia. All samples (n=28), including five controls, showed normal expression of actin, SRY and USP9Y. Sperm was not recovered from eight patients after testicular biopsy. Of these, four patients showed altered mRNA levels for the fertility genes, DAZ, RBMY1 and protamine-2. One patient, who was previously shown to be azoospermia factor region (AZF)b deleted, lacked RBM mRNA and presented with reduced amplification of protamine-2 mRNA. This correlated with previous studies, which proposed that RBM expression is exclusive to AZFb and that the lack of testicular RBMY1 mRNA results in suppressed spermatogenesis. Two patients were each lacking DAZ mRNA but did show expression of RBMY1 mRNA at a reduced level, suggesting that there might be residual spermatogenesis in the absence of DAZ expression. Protamine-2 mRNA was detected in one patient and was absent in the second patient. Finally, one patient lacked DAZ, RBMY1 and protamine-2 mRNA. The 19 remaining azoospermic patients presented with normal expression patterns for each of the fertility genes studied. This study demonstrates that the expression of spermatogenesis-specific genes varies in azoospermia. The study of the expression of such genes in a larger number of patients might be useful in characterizing and identifying subpopulations of azoospermic men.
authors.affiliation: National Diagnostics Centre, BioResearch Ireland, National University of Ireland, Galway, UK.
authors.forename: A
authors.initials: A
authors.lastname: Friel
authors.forename: J A
authors.initials: JA
authors.lastname: Houghton
authors.forename: M
authors.initials: M
authors.lastname: Glennon
authors.forename: R
authors.initials: R
authors.lastname: Lavery
authors.forename: T
authors.initials: T
authors.lastname: Smith
authors.forename: A
authors.initials: A
authors.lastname: Nolan
authors.forename: M
authors.initials: M
authors.lastname: Maher
mesh_terms: D001706:Biopsy; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics; D000075364:Deleted in Azoospermia 1 Protein; D005298:Fertility / Q000235:genetics; D005307:Fertilization in Vitro; D015870:Gene Expression; D006801:Humans; D007621:Karyotyping; D008297:Male; D009687:Nuclear Proteins; D009845:Oligospermia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D011479:Protamines / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000235:genetics*; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051876:Sex-Determining Region Y Protein; D013737:Testis / Q000378:metabolism* / Q000473:pathology; D014157:Transcription Factors; D014998:Y Chromosome
chemical_list: C095668:DAZ1 protein, human; D017931:DNA Primers; D004268:DNA-Binding Proteins; D000075364:Deleted in Azoospermia 1 Protein; D009687:Nuclear Proteins; D011479:Protamines; C501499:RBMY1A1 protein, human; D016601:RNA-Binding Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors; C068621:protamine 2