Nerve growth factor induces increased expression of a laminin-binding integrin in rat pheochromocytoma PC12 cells.
Rat pheochromocytoma PC12 cells exposed to nerve growth factor differentiate as sympathetic neurons and extend neurites on laminin and to a much lesser extent on fibronectin. Analysis of laminin fragments indicated that neurite outgrowth occurs mainly on fragment P1, corresponding to the center of the cross, and only poorly on fragment E8, a long arm structure that is active with other neuronal cells. Integrin antibodies prevented adhesion and neurite sprouting of these cells on laminin, fragment P1, and fibronectin. By affinity chromatography we isolated an integrin-type receptor for laminin consisting of two subunits with molecular massess of 180 and 135 kDa. The latter is recognized by an antiserum to integrin beta 1 subunit. The bound laminin receptor could be displaced by EDTA, but not by Arg-Gly-Asp or Tyr-Ile-Gly-Ser-Arg peptides. Affinity chromatography on laminin fragments showed that the 180/135 kDa receptor binds to P1. The expression of the 180-kDa alpha subunit of the laminin receptor at the cell surface was increased 10-fold after NGF treatment. The effect of NGF is specific since the amount of a 150-kDa fibronectin-binding integrin alpha subunit remained unchanged. Moreover, the increased expression of the 180/135 kDa receptor at the cell surface corresponded to a selective increase in cell adhesion to laminin and to fragment P1. The 180/135-kDa complex is thus an integrin-type receptor for laminin whose expression and binding specificity correlates with the capacity of NGF-induced PC12 cells to extend neurites on laminin.
Rossino
P
P
Dipartimento di Scienze Biomediche e Oncologia Umana, Italy.
Gavazzi
I
I
Timpl
R
R
Aumailley
M
M
Abbadini
M
M
Giancotti
F
F
Silengo
L
L
Marchisio
P C
PC
Tarone
G
G
D000310:Adrenal Gland Neoplasms; D000595:Amino Acid Sequence; D000818:Animals; D001369:Axons / Q000648:ultrastructure; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism; D016023:Integrins / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D009842:Oligopeptides / Q000494:pharmacology; D010673:Pheochromocytoma; D051381:Rats; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism*; D017474:Receptors, Laminin; D013564:Sympathetic Nervous System / Q000166:cytology; D014407:Tumor Cells, Cultured
D005353:Fibronectins; D016023:Integrins; D007797:Laminin; D009414:Nerve Growth Factors; D009842:Oligopeptides; D011971:Receptors, Immunologic; D017474:Receptors, Laminin; D004492:Edetic Acid