Chen Haiqi, Lui Wing-Yee, Mruk Dolores D, Xiao Xiang, Ge Renshan, Lian Qingquan, Lee Will M, Silvestrini Bruno, Cheng C Yan
The Mary M. Wohlford Laboratory for Male Contraceptive Research, Center for Biomedical Research, Population Council, New York, NY, USA.
School of Biological Sciences, University of Hong Kong, Pokfulam, Hong Kong, China.
Methods Mol Biol. 2018;1748:245-252. doi: 10.1007/978-1-4939-7698-0_17.
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 (CHNNaOS, 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 (CdCl) which is known to compromise the BTB integrity.
血睾屏障是哺乳动物睾丸中一种独特的超微结构，位于生精小管的基底膜附近，它将生精上皮分隔为基底室和近腔（顶端）室。血睾屏障除了限制生物分子（如旁分泌因子、激素）、水、电解质以及包括毒物和/或药物在内的其他物质通过细胞旁和跨细胞途径进入上皮的近腔室之外，还是支持精子发生的重要超微结构。因此，一种在体内监测其完整性的灵敏且可靠的检测方法有助于研究睾丸生物学。该检测方法基于完整的血睾屏障能够阻止小分子如磺基-NHS-LC-生物素（CHNNaOS，分子量556.59，一种水溶性且不能透过细胞膜的生物素化试剂）从生精上皮的基底室扩散到顶端室的能力。在此，我们总结了进行该检测的详细步骤，并获取半定量数据以评估与阳性对照相比血睾屏障受损的程度，阳性对照如用已知会破坏血睾屏障完整性的氯化镉（CdCl）处理大鼠。
