title: P-glycoprotein regulates blood-testis barrier dynamics via its effects on the occludin/zonula occludens 1 (ZO-1) protein complex mediated by focal adhesion kinase (FAK).
abstract: The blood-testis barrier (BTB), one of the tightest blood-tissue barriers in the mammalian body, creates an immune-privileged site for postmeiotic spermatid development to avoid the production of antibodies against spermatid-specific antigens, many of which express transiently during spermiogenesis and spermiation. However, the BTB undergoes extensive restructuring at stage VIII of the epithelial cycle to facilitate the transit of preleptotene spermatocytes and to prepare for meiosis. This action thus prompted us to investigate whether this stage can be a physiological window for the delivery of therapeutic and/or contraceptive drugs across the BTB to exert their effects at the immune-privileged site. Herein, we report findings that P-glycoprotein, an ATP-dependent efflux drug transporter and an integrated component of the occludin/zonula occludens 1 (ZO-1) adhesion complex at the BTB, structurally interacted with focal adhesion kinase (FAK), creating the occludin/ZO-1/FAK/P-glycoprotein regulatory complex. Interestingly, a knockdown of P-glycoprotein by RNAi was found to impede Sertoli cell BTB function, making the tight junction (TJ) barrier "leaky." This effect was mediated by changes in the protein phosphorylation status of occludin via the action of FAK, thereby affecting the endocytic vesicle-mediated protein trafficking events that destabilized the TJ barrier. However, the silencing of P-glycoprotein, although capable of impeding drug transport across the BTB and TJ permeability barrier function, was not able to induce the BTB to be "freely" permeable to adjudin. These findings indicate that P-glycoprotein is involved in BTB restructuring during spermatogenesis but that P-glycoprotein-mediated restructuring does not "open up" the BTB to make it freely permeable to drugs.
authors: The Mary M. Wohlford Laboratory for Male Contraceptive Research, Population Council, New York, NY 10065, USA.
authors: Linlin
authors: L
authors: Su
authors: Dolores D
authors: DD
authors: Mruk
authors: Wing-Yee
authors: WY
authors: Lui
authors: Will M
authors: WM
authors: Lee
authors: C Yan
authors: CY
authors: Cheng
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001814:Blood-Testis Barrier / Q000187:drug effects / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D006834:Hydrazines / Q000494:pharmacology; D015151:Immunoblotting; D007191:Indazoles / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D062793:Occludin; D010750:Phosphoproteins / Q000378:metabolism*; D061251:Primary Cell Culture; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D034622:RNA Interference; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
chemical_list: C432482:1-(2,4-dichlorobenzyl)indazole-3-carbohydrazide; D018435:ATP Binding Cassette Transporter, Subfamily B; D006834:Hydrazines; D007191:Indazoles; D008565:Membrane Proteins; D062793:Occludin; C568987:Ocln protein, rat; D010750:Phosphoproteins; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein; C095122:multidrug resistance protein 3; D051416:Focal Adhesion Protein-Tyrosine Kinases