Differential Regulation of Proline-rich Tyrosine Kinase 2/Cell Adhesion Kinase β (PYK2/CAKβ) and pp125FAK by Glutamate and Depolarization in Rat Hippocampus

doi:10.1074/jbc.271.46.28942

Differential Regulation of Proline-rich Tyrosine Kinase 2/Cell Adhesion Kinase β (PYK2/CAKβ) and pp125^FAK by Glutamate and Depolarization in Rat Hippocampus*

From INSERM U 114, Chaire de Neuropharmacologie, Collège de France, 11 place Marcelin Berthelot, 75231 Paris Cedex 05, France, the
^‡ Departamento de Histología, Facultad de Medicina, 11800 Montevideo, Uruguay and the
^¶ Department of Biochemistry, Cancer Research Institute, Sapporo Medical University, S-1 W-17, Sapporo 060, Japan

^∥To whom correspondence should be addressed. Tel.: 33-1-44-27-12-61; Fax: 33-1-44-27-12-75; E-mail: girault{at}infobiogen.fr.

Abstract

The mechanisms by which stimuli that raise cytosolic free Ca²⁺ concentrations in neurons can increase protein tyrosine phosphorylation are not known. Using rat hippocampal slices and cortical synaptosomes, we have examined the regulation of two highly related cytoplasmic tyrosine kinases, pp125 focal adhesion kinase (pp125^FAK) and proline-rich tyrosine kinase 2/cell adhesion kinase β (PYK2/CAKβ). Membrane depolarization increased tyrosine phosphorylation of PYK2/CAKβ and pp125^FAK. These effects were blocked by EGTA or by protein kinase C inhibitors (RO31-8220; GF109203X) and mimicked by ionomycin or phorbol 12-myristate 13-acetate, in the case of pp125^FAK, or their combination in the case of PYK2/CAKβ. Glutamate and specific agonists of ionotropic (α-amino-3-hydroxy-5-methyl-4-isoxazole propionate and N-methyl-D-aspartate) or metabotropic (trans-1-aminocyclopentane-1,3,-dicarboxylate) glutamate receptors stimulated the phosphorylation of pp125^FAK, but not of PYK2/CAKβ. Glutamate effects were prevented by GF109203X. Thus, in hippocampal slices, tyrosine phosphorylation of pp125^FAK and PYK2/CAKβ are regulated differentially by pathways involving Ca²⁺ and protein kinase C. pp125^FAK and PYK2/CAKβ may provide specific links between neuronal activity, increases in cytosolic Ca²⁺ and protein tyrosine phosphorylation, which may be important for neuronal survival, and synaptic plasticity.

Footnotes

↵^§ Supported by Comité Evaluation Orientation de la Coopération Scientifique and European Commission Contract ERBCI1*CT940038.
↵** Member of the Assistance Publique des Hôpitaux de Paris.
↵* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

LTP

long term potentiation

ACSF

artificial cerebrospinal fluid

AMPA

α-amino-3-hydroxy-5-methyl-4-isoxazole propionate

CAKβ

cell adhesion kinase β

pp125^FAK

pp125 focal adhesion kinase

LTD

long term depression

MAP kinase

mitogen-activated protein kinase

NMDA

N-methyl-D-aspartate

PKC

protein kinase C

PMA

phorbol 12-myristate 13-acetate

PYK2

proline-rich tyrosine kinase 2

SH2

Src-homology domain 2

SH3

Src-homology domain 3

t-ACPD

trans-1-aminocyclopentane-1,3,-dicarboxylate

TTX

tetrodotoxin.
↵²J. C. Siciliano, P. Derkinderen, M. Toutant, and J.-A. Girault, manuscript in preparation.
- Received September 23, 1996.