Expression Of CR16 In Testicular Tissue And The Initial Exploration Of Its Role In Pathogenesis Of Azoospermia

PartⅠThe expression of CR16 in testicular tissue of normal fertile Sprague-Dwaley ratsObjective To investigate the expression of Corticosteroids and regional expression 16(CR16)in testicular tissue of normal fertile Sprague-Dwaley(SD)rats and provide an initial animal experiments and methods preparation for further study of CR16 in the human reproductive system.Methods Immunohistochemical was used to measure the levels of CR16 protein in the testes of 5 normal fertile SD rats.Results Immunohistochemical showed that CR16 protein expressed in the Sertoli cells and Sertoli cell-spermatid junction(sSspJs)in epithelial of seminiferous tubules.Conclusion The CR16 protein expressed in the Sertoli cells and SspJs in testicular tissue of normal fertile SD rats, which indicates that CR16 may have relationship with spermatogenesis. PartⅡThe expression of CR16 in testicular tissue of azoospermia patients and healthy menObjective To investigate the expression of CR16 in the testes of patients with non-obstructive azoospermia, obstructive azoospermia and healthy men and discuss its correlation with azoospermia.Methods Immunohistochemical and reverse transcript polymerase chain reaction (RT-PCR) were used to measure the levels of CR16 protein and mRNA in the testes of 6 patients with non-obstructive azoospermia, 23 patients with obstructive azoospermia and 6 healthy men.Results Immunohistochemical showed that CR16 protein expressed in the cytoplasm and membrane of Sertoli cells and SspJs in epithelial of seminiferous tubules. And the level of CR16 protein was markedly lower in non-obstructive azoospermia patients than obstructive azoospermia patients and healthy men. RT-PCR showed that the level of CR16 mRNA was significantly decreased in non-obstructive azoospermia patients(0.640±0.021)compared with the healthy men(0.952±0.036)(P<0.01), and had no statistical significance between obstructive azoospermia patients(0.953±0.036)and healthy men(P>0.05).Conclusion The levels of CR16 protein and mRNA decrease in the testes of non-obstructive azoospermia patients, which indicates that CR16 may have relationship with pathogenesis of azoospermia and participate in spermatogenesis dysfunction. Objective To determine the expression of neural Wiscott-Aldrich syndrome protein (N-WASP) in the testes of patients with non-obstructive azoospermia and healthy men and discuss its correlation with CR16 in humen testes and their role in spermatogenesis.Method Immunohistochemical and RT-PCR were used to measure the levels of N-WASP protein and mRNA in the testes of 6 patients with non-obstructive azoospermia and 6 healthy men. Double-staining immunofluorescence and laser scanning confocal microscopy (LSCM) were used to demonstrate co-localization of CR16 and N-WASP proteins.Results Immunohistochemical showed that N-WASP protein expressed in the cytoplasm and membrane of Sertoli cells and SspJs in epithelial of seminiferous tubules. And the level of N-WASP protein was markedly lower in non-obstructive azoospermia patients than healthy men. RT-PCR showed that the level of N-WASP mRNA was significantly decreased in non-obstructive azoospermia patients(0.645±0.025)compared with the healthy men(0.960±0.031)(P<0.01). The ratio of CR16 and N-WASP mRNA was approximately 1:1 in both groups of 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 N-WASP-CR16 protein complex.Conclusion The decreased levels of CR16 and N-WASP in the testes of men with non-obstructive azoospermia compared with healthy men and the co-localization at SspJs as a N-WASP-CR16 protein complex in the epithelium of seminiferous tubules suggests a role in the pathogenesis of non-obstructive azoospermia.