Interactions of Sertoli cells with laminin are essential to maintain integrity of the cytoskeleton and barrier functions of cells in culture in the two-chambered assembly.
The addition of anti-laminin IgG to the basal surfaces of rat Sertoli cells in culture in a two-chambered assembly results in a perturbation of F-actin arrangements, including disruption of the pericellular circumferal rings, impairments of the Sertoli cell permeability barrier, and subsequently focal defoliation, followed by cell reaggregation. The pentapeptide YIGSR, which competes with the laminin receptor for laminin (Kleinman and Weeks: Curr. Oph. Cell Biol., 1:964-967, 1989; Graf et al.: Biochemistry, 26:6896-6900, 1987) also elicited focal defoliation of Sertoli cells from the extracellular matrix-coated filter in the two-chambered assembly. Addition of YIGSR to Sertoli cell cultures resulted in cell detachment within 2 to 3 h. In contrast, the irrelevant peptide YIGSE had no detectable effects. The anti-laminin IgG was effective only when added to the chamber in which access was readily available to the basal surfaces of Sertoli cells, but YIGSR was effective when added either to the outer chamber or to the inner chamber. These data were interpreted to indicate that the Sertoli cell barrier generated in the two-chambered assembly allowed a relatively rapid diffusion of YIGSR between chambers, but prevented the rapid equilibration of anti-laminin IgG between compartments. Addition of anti-laminin IgG to the basal, but not to the apical surfaces of Sertoli cells, resulted in more rapid rates of equilibration of [3H]-methoxyinulin and [86Rb]Cl across the Sertoli cell monolayer. This evidence of impairment to the integrity of the barrier was detected prior to the disruption of stress fibers and focal defoliation, but after evidence of dissolution of the circumferal F-actin ring, which occurred within 1 h after addition of anti-laminin IgG. We consider the possibility that a transmembrane link exists between extracellular laminin and cytoskeletal elements which modulates the circumferal F-actin ring. We further postulate that this linkage can influence the nature of tight junctional complexes, and thereby the integrity of the Sertoli cell barrier.
Tung
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PS
Banting and Best Department of Medical Research, C.H. Best Institute, University of Toronto, Ontario, Canada.
Fritz
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IB
D000199:Actins / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002462:Cell Membrane / Q000648:ultrastructure; D016764:Cell Polarity; D003599:Cytoskeleton / Q000648:ultrastructure; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D007797:Laminin / Q000502:physiology*; D008297:Male; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D010539:Permeability; D051381:Rats; D017208:Rats, Wistar; D017474:Receptors, Laminin / Q000502:physiology*; D012708:Sertoli Cells / Q000166:cytology* / Q000378:metabolism
D000199:Actins; D007797:Laminin; D009842:Oligopeptides; D017474:Receptors, Laminin