Monitoring the Integrity of the Blood-Testis Barrier (BTB): An In Vivo Assay.
Monitoring the Integrity of the Blood-Testis Barrier (BTB): An In Vivo Assay.
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DOI：
10.1007/978-1-4939-7698-0_17
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Cheng CY
中科院分区：
文献类型：
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作者：
Chen H;Lui WY;Mruk DD;Xiao X;Ge R;Lian Q;Lee WM;Silvestrini B;Cheng CY
血-睾丸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，该基底膜将生精上皮分为基底室和腺体(顶端)室。除了限制生物分子(如旁分泌因子、激素)、水、电解质和其他物质(包括毒物和/或药物)的细胞旁和跨细胞通道进入上皮腺室外，BTB是支持精子发生的重要超微结构。因此，一种灵敏可靠的方法来监测其在体内的完整性有助于研究睾丸生物学。这种分析是基于完整的btb从生精上皮的底部到顶端隔间排除小分子扩散的能力，如磺基-NHS-LC-生物素(C20H29N4NaO9S2，Mr 556.59，一种水溶性和膜不渗透的生物素化试剂)。在这里，我们总结了进行检测的详细程序，并获得半定量数据，以评估与阳性对照相比BTB损伤的程度，例如用已知损害BTB完整性的氯化镉(CDCl2)处理大鼠。
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.