An Occludin-Focal Adhesion Kinase Protein Complex at the Blood-Testis Barrier: A Study Using the Cadmium Model

• Published in: Endocrinology (Philadelphia) 2009-07, Vol.150 (7), p.3336-3344
• Main Authors: Siu, Erica R, Wong, Elissa W. P, Mruk, Dolores D, Sze, K. L, Porto, Catarina S, Cheng, C. Yan
• Format: Article
• Language: English
• Subjects: Actin; Adhesion; Animals; Biological and medical sciences; Blood; Blood-Testis Barrier - drug effects; Blood-Testis Barrier - physiology; Cadmium; Cadmium Chloride - pharmacology; Cell adhesion; Connexin 43; Cytoskeleton; Epithelium; Fluorescence microscopy; Focal adhesion kinase; Focal Adhesion Kinase 1 - metabolism; Fundamental and applied biological sciences. Psychology; Gametocytes; Immunohistochemistry; In vivo methods and tests; Internalization; Kinases; Male; Membrane proteins; Membrane Proteins - metabolism; Models, Biological; N-Cadherin; Occludin; Permeability - drug effects; Phosphoproteins - metabolism; Proteins; Rats; Rats, Sprague-Dawley; Sertoli cells; Sertoli Cells - drug effects; Sertoli Cells - metabolism; Spermatocytes; Testes; Tight Junctions - drug effects; Tight Junctions - physiology; Vertebrates: endocrinology; Zonula Occludens-1 Protein
• ISSN: 0013-7227; 1945-7170
• DOI: 10.1210/en.2008-1741

Several integral membrane proteins that constitute the blood-testis barrier (BTB) in mammalian testes, in particular rodents, are known to date. These include tight junction (TJ) proteins (e.g. occludin, junctional adhesion molecule-A, claudins), basal ectoplasmic specialization proteins (e.g. N-cadherin), and gap junction proteins (e.g. connexin43). However, the regulators (e.g. protein kinases and phosphatases) that affect these proteins, such as their interaction with the cytoskeletal actin, which in turn confer cell adhesion at the TJ, remain largely unknown. We report herein that focal adhesion kinase (FAK) is a putative interacting partner of occludin, but not claudin-11 or junctional adhesion molecule-A. Immunohistochemistry and fluorescence microscopy studies illustrated that the expression of FAK in the seminiferous epithelium of adult rat testes was stage specific. FAK colocalized with occludin at the BTB in virtually all stages of the seminiferous epithelial cycle but considerably diminished in stages VIII–IX, at the time of BTB restructuring to facilitate the transit of primary leptotene spermatocytes. Using Sertoli cells cultured in vitro with established TJ-permeability barrier and ultrastructures of TJ, basal ectoplasmic specialization and desmosome-like junction that mimicked the BTB in vivo, FAK was shown to colocalize with occludin and zonula occludens-1 (ZO-1) at the Sertoli-Sertoli cell interface. When these Sertoli cell cultures were treated with CdCl2 to perturb the TJ-barrier function, occludin underwent endocytic-mediated internalization in parallel with FAK and ZO-1. Thus, these findings demonstrate that FAK is an integrated regulatory component of the occludin-ZO-1 protein complex, suggesting that functional studies can be performed to study the role of FAK in BTB dynamics. Focal adhesion kinase (FAK) is an integrated regulatory component of the occludin-ZO-1 protein complex, suggesting functional studies can be performed to study the role of FAK in blood-testis barrier dynamics.