Gap junction formation between cultured embryonic lens cells is inhibited by antibody to N-cadherin.
Intercellular communication mediated by gap junctions is important for tissue homeostasis in the avascular lens, and extensive areas of gap junctions form between fiber cells during fiber cell differentiation and lens development. We examined the role of the calcium-dependent cell adhesion molecule, N-cadherin, in the process of gap junction formation between fiber cells. Lentoids, multicellular structures with characteristics of differentiated fiber cells, were isolated from embryonic chick lens cultures and subsequently paired to provide an in vitro model of fiber cell interactions. Gap junction formation between cells of paired lentoids was monitored by observing the lentoid-to-lentoid transfer of fluorescent dyes, either calcein or Lucifer yellow, over a time course of up to 48 hr. Dye transfer between lentoids was inhibited upon the addition to the medium of Fab fragments (100-622 microgram/ml) of a monoclonal antibody specific for N-cadherin, and also by the reduction of extracellular calcium in the incubation medium. However, the addition of Fab fragments (100-1500 microgram/ml) of an antibody to a fiber-cell-specific integral membrane protein, MIP, did not change the time course nor extent of dye transfer between lentoids. Our results, using cultured embryonic cells, extend those from previous studies with cell lines and transfected cells. We conclude that cadherin interactions facilitate the formation of gap junctions between embryonic lens fiber cells, by the stabilization of membrane appositions and/or by the generation of an intracellular signal(s).
Frenzel
E M
EM
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108, USA. frenz003@maroon.tc.umn.edu
Johnson
R G
RG
D000818:Animals; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000276:immunology / Q000378:metabolism*; D015820:Cadherins / Q000276:immunology* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D002642:Chick Embryo; D005452:Fluoresceins / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D017629:Gap Junctions / Q000378:metabolism*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007908:Lens, Crystalline / Q000196:embryology* / Q000378:metabolism; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D015820:Cadherins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007140:Immunoglobulin Fab Fragments; D002118:Calcium; C007740:fluorexon