Monitoring the Integrity of the Blood-Testis Barrier (BTB): An In Vivo Assay
- 763 Downloads
The blood-testis barrier is a unique ultrastructure in the mammalian testis, located near the basement membrane of the seminiferous tubule that segregates the seminiferous epithelium into the basal and the adluminal (apical) compartment. Besides restricting paracellular and transcellular passage of biomolecules (e.g., paracrine factors, hormones), water, electrolytes, and other substances including toxicants and/or drugs to enter the adluminal compartment of the epithelium, the BTB is an important ultrastructure that supports spermatogenesis. As such, a sensitive and reliable assay to monitor its integrity in vivo is helpful for studying testis biology. This assay is based on the ability of an intact BTB to exclude the diffusion of a small molecule such as sulfo-NHS-LC-biotin (C20H29N4NaO9S2, Mr. 556.59, a water-soluble and membrane-impermeable biotinylation reagent) from the basal to the apical compartment of the seminiferous epithelium. Herein, we summarize the detailed procedures on performing the assay and to obtain semiquantitative data to assess the extent of BTB damage when compared to positive controls, such as treatment of rats with cadmium chloride (CdCl2) which is known to compromise the BTB integrity.
KeywordsTestis Blood-testis barrier Spermatogenesis Sertoli cells Tight junction Ectoplasmic specialization Actin filaments Assay
This work was supported by grants from the National Institutes of Health, NICHD R01 HD056034 to C.Y.C., and U54 HD029990 Project 5 to C.Y.C.; Hong Kong Research Grants Council (RGC)/National Natural Science Foundation of China Joint Research Scheme (N_HKU 717/12) to W.M.L., Hong Kong RGC Grant GRF17100816 to W.Y.L. and GRF 771513 to W.M.L., Hong Kong University Seed Funding to W.Y.L. and W.M.L.; NSFC Grant 81730042 to R.S.G. and NSFC Grant 31371176 to X.X., and Zhejiang Province Department of Science Technology Funding 2016F10010 to X.X.; H.C. was supported in part by The S.Y. Law Memorial Fellowship and The F. Lau Memorial Fellowship.