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.
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This work was supported by a grant from the National “Tenth Five Years” Key Technologies R&D Programme, China (no. 2004BA720A33-01).
Prof. C. Xiong directed this investigation, Dr. Y. Zhang greatly supported the experimental aspects of this work, Zina Wen and Lian Hu made extensive contributions to the study, and Wenjuan Pang was responsible for proofreading of the text.
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Xiang, W., Wen, Z., Pang, W. et al. CR16 forms a complex with N-WASP in human testes. Cell Tissue Res 344, 519–526 (2011). https://doi.org/10.1007/s00441-011-1159-9
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DOI: https://doi.org/10.1007/s00441-011-1159-9