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pp60c-src Is a Negative Regulator of Laminin-1-Mediated Neurite Outgrowth in Chick Sensory Neurons

https://doi.org/10.1006/mcne.2002.1157Get rights and content

Abstract

Multiple protein tyrosine kinases regulate neurite outgrowth in the developing nervous system. To begin to unravel the complexity of this regulation, we addressed the role of one specific kinase, pp60c-src, in chick dorsal root ganglion (DRG) neurons grown on laminin-1, a well-characterized system to study neurite outgrowth. Pharmacological inhibition of all tyrosine kinases by genestein treatment of chick DRG neurons significantly increased neurite number and length by ∼50%. Similar increases in these parameters occurred when src-family kinases were inhibited using PP2. To implicate pp60c-src directly in neurite outgrowth, we inactivated it in DRG neuronal growth cones using Chromophore-Assisted Laser Inactivation (CALI). CALI of pp60c-src resulted in an 85% inactivation of its kinase activity and a 63% reduction in phosphotyrosine immunofluorescence in neurons. Microscale CALI of pp60c-src in DRG growth cones caused a significant and acute two-fold increase in neurite extension rate during irradiation. These findings demonstrate that pp60c-src is a negative regulator of laminin-1-mediated neurite outgrowth in chick sensory neurons.

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    1

    These two authors shared equally in the work for this study.

    2

    Present address: Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Box G-B393, Providence, RI 02912

    3

    To whom correspondence and reprint requests should be addressed. Fax: 617-636-0445. E-mail: [email protected].

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