Organization of point contacts in neuronal growth cones.
Growth cones from rat dorsal root ganglia plated on laminin contain integrin clusters over the entire growth cone surface, and growth cones make transient adhesions at sites called point contacts. We examined, by immunocytochemistry and confocal microscopy, the composition and distribution of point contacts in neuronal growth cones. Vinculin was concentrated in the central domain of growth cones and at the tips of filopodia. Vinculin was specifically associated with integrin clusters at the membrane-substrate interface and thus marked point contacts. The cytoskeletal proteins paxillin and talin colocalized with beta1 integrin in a subpopulation of clusters restricted to the central domain of the growth cone and to the tips of filopodia. The neuron-specific kinase, FAK+ also distributed with the vinculin-positive clusters. The Rho family proteins RhoA, RhoB, and Cdc42 were present in growth cones, and a few Rho clusters were colocalized with vinculin. Examination of proteins resistant to detergent extraction in PC12 cells confirmed the retention of beta1 integrin, paxillin, talin, and vinculin with the cytoskeleton. Moreover, we detected FAK+ and RhoA in the detergent-resistant cytoskeleton, supporting their distribution to point contacts. Our observations indicate that two types of integrin clusters are present in growth cones: those associated with vinculin at the cell substratum interface, and those not associated with vinculin. Point contacts are mature adhesion sites defined by the presence of both beta1 integrin and vinculin, and they are associated with signaling proteins.
Renaudin
A
A
Département de Pathologie et Biologie Cellulaire, Université de Montréal, Montréal, Québec, Canada.
Lehmann
M
M
Girault
J
J
McKerracher
L
L
D000818:Animals; D001923:Brain Chemistry; D016466:CHO Cells / Q000737:chemistry; D002478:Cells, Cultured; D006224:Cricetinae; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology*; D020439:Growth Cones / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000032:analysis; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D012583:Schwann Cells / Q000737:chemistry / Q000166:cytology; D013499:Surface Properties; D016608:Talin / Q000378:metabolism; D016596:Vinculin / Q000032:analysis*; D020741:rho GTP-Binding Proteins / Q000378:metabolism
D003598:Cytoskeletal Proteins; D019012:Integrin beta1; D051419:Paxillin; D010750:Phosphoproteins; C496063:Pxn protein, rat; D016608:Talin; D016596:Vinculin; D011505:Protein-Tyrosine Kinases; D020741:rho GTP-Binding Proteins