title: CR16 forms a complex with N-WASP in human testes.
abstract: The expression of corticosteroids and regional expression 16 (CR16) and neural Wiscott-Aldrich syndrome protein (N-WASP) was studied in the testes of men with idiopathic azoospermia by (1) immunohistochemistry and Western blot to detect the expression of CR16 and N-WASP protein, (2) reverse transcription with the polymerase chain reaction (RT-PCR) to determine the amounts of CR16 and N-WASP mRNA, and (3) double-staining immunofluorescence and laser scanning confocal microscopy (LSCM) to demonstrate the co-localization of CR16 and N-WASP proteins. Immunohistochemistry revealed CR16 and N-WASP proteins in the Sertoli cells, specifically in the Sertoli cell-spermatogenic cell junctions (SspJs) in the epithelium of the seminiferous tubules. RT-PCR indicated that the relative amounts of CR16 and N-WASP mRNA were significantly decreased in men with idiopathic azoospermia, although the ratio of CR16 and N-WASP mRNA was approximately 1:1 in both groups of men. Western blot showed the presence of the CR16 monomer and N-WASP in all tissues examined, but the relevant bands were paler in men with idiopathic azoospermia than those in healthy men. The distribution of CR16 and N-WASP was similar in the epithelium of seminiferous tubules based on double-staining immunofluorescence and LSCM, thereby suggesting that CR16 and N-WASP were co-localized in the SspJs as a CR16/N-WASP complex. The decreased levels of CR16 and N-WASP in the testes of men with idiopathic azoospermia compared with healthy men and the co-localization at the SspJs as a CR16/N-WASP complex in the epithelium of seminiferous tubules suggest that CR16 and N-WASP play a role in the pathogenesis of azoospermia.
authors.affiliation: Family Planning Research Institute, Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People's Republic of China.
authors.forename: Wenpei
authors.initials: W
authors.lastname: Xiang
authors.forename: Zina
authors.initials: Z
authors.lastname: Wen
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Pang
authors.forename: Lian
authors.initials: L
authors.lastname: Hu
authors.forename: Chengliang
authors.initials: C
authors.lastname: Xiong
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000328:Adult; D053713:Azoospermia / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D015153:Blotting, Western; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism*; D011446:Prospective Studies; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000378:metabolism; D013737:Testis / Q000378:metabolism* / Q000473:pathology; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000235:genetics / Q000378:metabolism*; D055815:Young Adult
chemical_list: D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D012333:RNA, Messenger; C562194:WIPF3 protein, human; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal