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Caveolin isoform expression during differentiation of C6 glioma cells
Author(s) -
Silva W.I.,
Maldonado H.M.,
Velázquez G.,
RubioDávila M.,
Miranda J.D.,
Aquino E.,
Mayol N.,
CruzTorres A.,
Jardón J.,
SalgadoVillanueva I.K.
Publication year - 2005
Publication title -
international journal of developmental neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.761
H-Index - 88
eISSN - 1873-474X
pISSN - 0736-5748
DOI - 10.1016/j.ijdevneu.2005.07.007
Subject(s) - caveolae , caveolin , biology , gene isoform , microbiology and biotechnology , caveolin 1 , endocytosis , mertk , caveolin 3 , signal transduction , gene , receptor tyrosine kinase , biochemistry , receptor
Caveolae, a specialized form of lipid rafts, are cholesterol‐ and sphingolipid‐rich membrane microdomains implicated in potocytosis, endocytosis, transcytosis, and as platforms for signal transduction. One of the major constituents of caveolae are three highly homologous caveolin isoforms (caveolin‐1, caveolin‐2, and caveolin‐3). The present study expands the analysis of caveolin isoform expression in C6 glioma cells. Three complementary approaches were used to assess their differential expression during the dibutyryl‐cyclic AMP‐induced differentiation of C6 cells into an astrocyte‐like phenotype. Immunoblotting, conventional RT‐PCR, and real‐time RT‐PCR analysis established the expression of the caveolin‐3 isoform in C6 cells, in addition to caveolin‐1 and caveolin‐2. Similar to the other isoforms, caveolin‐3 was associated with light‐density, detergent‐insoluble caveolae membrane fractions obtained using sucrose‐density gradient centrifugation. The three caveolin isoforms display different temporal patterns of mRNA/protein expression during the differentiation of C6 cells. Western blot and real‐time RT‐PCR analysis demonstrate that caveolin‐1 and caveolin‐2 are up‐regulated during the late stages of the differentiation of C6 cells. Meanwhile, caveolin‐3 is gradually down‐regulated during the differentiation process. Indirect immunofluorescence analysis via laser‐scanning confocal microscopy reveals that the three caveolin isoforms display similar subcellular distribution patterns. In addition, co‐localization of caveolin‐1/caveolin‐2 and caveolin‐1/caveolin‐3 was detected in both C6 glioma phenotypes. The findings reveal a differential temporal pattern of caveolin gene expression during phenotypic differentiation of C6 glioma cells, with potential implications to developmental and degenerative events in the brain.