Activation of the somatostatin receptor sst2, a member of the G i protein-coupled receptor family, results in the stimulation of a protein-tyrosine phosphatase activity involved in the sst2-mediated growth inhibitory signal. Here, we report that SHP-1, a cytoplasmic proteintyrosine phosphatase containing two Src homology 2 domains constitutively associated with sst2 as evidence by coprecipitation of SHP-1 protein with sst2, in Chinese hamster ovary cells coexpressing sst2 and SHP-1. Activation of sst2 by somatostatin resulted in a rapid dissociation of SHP-1 from sst2 accompanied by an increase of SHP-1 activity. SHP-1 was phosphorylated on tyrosine in control cells and somatostatin induced a rapid and transient dephosphorylation on tyrosine residues of the enzyme. Stimulation of SHP-1 activity by somatostatin was abolished by pertussis toxin pretreatment of cells. G i␣3 was specifically immunoprecipitated by anti-sst2 and anti-SHP-1 antibodies, and somatostatin induced a rapid dissociation of G i␣3 from sst2, suggesting that G i␣3 may be involved in the sst2⅐SHP-1 complexes. Finally, somatostatin inhibited the proliferation of cells coexpressing sst2 and SHP-1, and this effect was suppressed in cells coexpressing sst2 and the catalytic inactive SHP-1 (C453S mutant). Our data identify SHP-1 as the tyrosine phosphatase associated with sst2 and demonstrate that this enzyme may be an initial key transducer of the antimitogenic signaling mediated by sst2.
Hybrid cell lines derived from neonatal rat dorsal root ganglia neurons fused with the mouse neuroblastoma N18Tg2 exhibit sensory neuron-like properties not displayed by the parental neuroblastoma. These properties include an inward (depolarizing) current with a conductance increase in response to activation of a bradykinin receptor, an inward (depolarizing) current with a conductance increase in response to the sensory excitotoxin capsaicin, the expression of sensory neuropeptides (substance P, CGRP and somatostatin), the expression of phosphatidylinositol-anchored molecules including adhesion molecules of the immunoglobulin superfamily that can be regulated in serum-free culture by nerve growth factor (N-CAM, F-3 and Thy-1), and low permissivity to herpes simplex virus infection. These lines thus provide appropriate models for the study of mechanisms involved in nociceptor activation and the regulation of expression of sensory-neuron specific markers including neuropeptides.
In the central nervous system, postmitotic neurons migrate along astrocytic processes to reach their adult position. The molecular mechanisms of this guided migration are not clearly defined, although some steps have been shown to involve proteases and cell adhesion molecules. We report that monovalent antibodies (Fab fragments) raised against an endogenous cerebellar soluble lectin (CSL) completely inhibit neuronal migration in cultures of cerebellar explants at concentrations as low as 50 ,ug/ml. A similar inhibition pattern was obtained with Fab fragments prepared against one of the endogenous glycoprotein ligands of CSL, the 31-kDa glycoprotein (this glycoprotein is a membrane-bound glycoprotein specifically occurring, in the cerebellum, at the surface of immature neurons). We propose that this lectin-glycoprotein interaction supports the adhesion between neurons and the astrocyte guide during the migration of cerebellar immature neurons.
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