Monitoring the integrity of the blood-testis barrier (BTB): An in vivo assay

Haiqi Chen, Wing yee Lui, Dolores D. Mruk, Xiang Xiao, Renshan Ge, Qingquan Lian, Will M. Lee, Bruno Silvestrini, C. Yan Cheng

Research output: Chapter in Book/Report/Conference proceedingChapter

16 Scopus citations


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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages8
StatePublished - 2018
Externally publishedYes

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Actin filaments
  • Assay
  • Blood-testis barrier
  • Ectoplasmic specialization
  • Sertoli cells
  • Spermatogenesis
  • Testis
  • Tight junction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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