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Title

PC12 cells have caveolae that contain TrkA

Other TitlesCaveolae-disrupting drugs inhibit nerve growth factor-induced, but not epidermal growth factor-induced, MAPK phosphorylation
AuthorsMartín-Zanca, Dionisio
Issue Date2000
PublisherAmerican Society for Biochemistry and Molecular Biology
CitationJournal of Biological Chemistry 275(48): 37846-37852 (2000)
AbstractNerve growth factor (NGF) induces survival and differentiation of the neural crest-derived PC12 cell line. Caveolae are cholesterol-enriched, caveolin-containing plasma membrane microdomains involved in vesicular transport and signal transduction. Here we demonstrate the presence of caveolae in PC12 cells and their involvement in NGF signaling. Our results showed the expression of caveolin-1 by Western blot and confocal immuno-microscopy. The presence of plasma membrane caveolae was directly shown by rapid-freeze deep-etching electron microscopy. Moreover, combined deepetching and immunogold techniques revealed the presence of the NGF receptor TrkA in the caveolae of PC12 ceils. These data together with the cofractionation of Shc, Ras, caveolin, and TrkA in the caveolae fraction supported a role for these plasma membrane microdomains in NGF signaling. To approach this hypothesis, caveolae were disrupted by treatment of PC12 cells with cholesterol binding drugs. Either filipin or cyclodextrin treatment increased basal levels of MAPK phosphorylation. In contrast, pretreatment of PC12 cells with these drugs inhibited the NGF- but not the epidermal growth factor-lnduced MAPK phosphorylation without affecting the TrkA autophosphorylation. Taken together, our results demonstrate the presence of caveolae in PC12 ceils, which contain the high affinity NGF receptor TrkA, and the specific involvement of these cholesterol-enriched plasma membrane microdomains in the propagation of the NGF-induced signal.
URIhttp://hdl.handle.net/10261/66721
DOI10.1074/jbc.M000487200
Identifiersdoi: 10.1074/jbc.M000487200
issn: 0021-9258
e-issn: 1083-351X
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