TY - DATA T1 - Isoform-Level Gene Expression Profiles of Human Y Chromosome Azoospermia Factor Genes and Their X Chromosome Paralogs in the Testicular Tissue of Non-Obstructive Azoospermia Patients PY - 2015/09/04 AU - Diba Ahmadi Rastegar AU - Mehdi Sharifi Tabar AU - Mehdi Alikhani AU - Pouria Parsamatin AU - Fazel Sahraneshin Samani AU - Marjan Sabbaghian AU - Mohammad Ali Sadighi Gilani AU - Ali Mohammad Ahadi AU - Anahita Mohseni Meybodi AU - Abbas Piryaei AU - Naser Ansari-Pour AU - Hamid Gourabi AU - Hossein Baharvand AU - Ghasem Hosseini Salekdeh UR - https://acs.figshare.com/articles/journal_contribution/Isoform_Level_Gene_Expression_Profiles_of_Human_Y_Chromosome_Azoospermia_Factor_Genes_and_Their_X_Chromosome_Paralogs_in_the_Testicular_Tissue_of_Non_Obstructive_Azoospermia_Patients/2135281 DO - 10.1021/acs.jproteome.5b00520.s001 L4 - https://ndownloader.figshare.com/files/3769123 KW - WB KW - 5D KW - 5C KW - 41 alternative transcripts KW - PRY KW - CDY KW - USP 9Y DDX 3Y XKRY KW - HSFY KW - SCOS KW - IHC KW - PCR KW - Human Y Chromosome Azoospermia Factor Genes KW - NOA KW - sperm retrieval KW - 28 premiotic maturation arrest KW - expression KW - X Chromosome Paralogs KW - AZFc region genes KW - 1A KW - RBMX KW - KDM KW - X chromosome homologue transcripts KW - CYORF KW - 12 MA patients KW - Y chromosome genes KW - TESE KW - EIF KW - BPY KW - isoform level signature KW - testicular sperm extraction KW - RPS KW - 15B KW - 4Y KW - spermatogenesi KW - protein level KW - X chromosome counterparts KW - DAZL 1 transcripts KW - 1AY KW - testis cell types KW - RBMY KW - azoospermic men KW - 15A N2 - The human Y chromosome has an inevitable role in male fertility because it contains many genes critical for spermatogenesis and the development of the male gonads. Any genetic variation or epigenetic modification affecting the expression pattern of Y chromosome genes may thus lead to male infertility. In this study, we performed isoform-level gene expression profiling of Y chromosome genes within the azoospermia factor (AZF) regions, their X chromosome counterparts, and few autosomal paralogues in testicular biopsies of 12 men with preserved spermatogenesis and 68 men with nonobstructive azoospermia (NOA) (40 Sertoli-cell-only syndrome (SCOS) and 28 premiotic maturation arrest (MA)). This was undertaken using quantitative real-time PCR (qPCR) at the transcript level and Western blotting (WB) and immunohistochemistry (IHC) at the protein level. We profiled the expression of 41 alternative transcripts encoded by 14 AZFa, AZFb, and AZFc region genes (USP9Y, DDX3Y, XKRY, HSFY1, CYORF15A, CYORF15B, KDM5D, EIF1AY, RPS4Y2, RBMY1A1, PRY, BPY2, DAZ1, and CDY1) as well as their X chromosome homologue transcripts and a few autosomal homologues. Of the 41 transcripts, 18 were significantly down-regulated in men with NOA when compared with those of men with complete spermatogenesis. In contrast, the expression of five transcripts increased significantly in NOA patients. Furthermore, to confirm the qPCR results at the protein level, we performed immunoblotting and IHC experiments (based on 24 commercial and homemade antibodies) that detected 10 AZF-encoded proteins. In addition, their localization in testis cell types and organelles was determined. Interestingly, the two missing proteins, XKRY and CYORF15A, were detected for the first time. Finally, we focused on the expression patterns of the significantly altered genes in 12 MA patients with successful sperm retrieval compared to those of 12 MA patients with failed sperm retrieval to predict the success of sperm retrieval in azoospermic men. We showed that HSFY1-1, HSFY1-3, BPY2-1, KDM5C2, RBMX2, and DAZL1 transcripts could be used as potential molecular markers to predict the presence of spermatozoa in MA patients. In this study, we have identified isoform level signature that can be used to discriminate effectively between MA, SCOS, and normal testicular tissues and suggests the possibility of diagnosing the presence of mature sperm cell in azoospermic men to prevent additional testicular sperm extraction (TESE) surgery. ER -