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Expression Changes of P2Y2 and P2Y6 Nucleotide Receptors During C6 Glioma Cell Differentiation
Author(s) -
Garcia Jose O.,
Anglero Yesseinia,
Cruz Alexandra,
Aquino Edna E.,
Gonzalez Fernando A.,
Silva Walter I.
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1211-a
Subject(s) - receptor , p2y receptor , biology , microbiology and biotechnology , nucleotide , purinergic receptor , biochemistry , gene
Nucleotide receptors, which are activated by ATP, UTP and their metabolites, can induce a diversity of physiological responses in most tissues. Studies provide evidence of nucleotide signaling roles in the nervous system. Nevertheless, little is known about nucleotide receptors function during glial differentiation. Using C6 glioma cells, we studied the expression of four P2Y receptor subtypes: P2Y 1 , P2Y 2 , P2Y 6 , and P2Y 12 , during cellular differentiation. A nucleotide receptors time course was prepared using real‐time RT‐PCR and immunoblotting. In addition, sucrose density gradient fractionation with Na 2 CO 3 was executed for P2Y 2 and P2Y 6. Laser scanning confocal microscopy was used to determine P2Y receptors expression. Time course analysis results indicate that P2Y 2 receptors are down‐regulated opposed to P2Y 6 which shows a slightly up‐regulation. P2Y 2 and P2Y 6 detected between fractions 3–5, which are characterized to be caveolar microdomains fractions. Indirect immunofluorescence analysis of P2Y 2 and caveolin‐1 in undifferentiated cells shows co‐localization of them; a result confirmed using C6 glioma cells transfected with a cav1‐EGFP chimera. Nucleotide receptors expression may be correlated to astrogliosis process observed during physiological and pathophysiological responses in nervous system. Supported by NIH GM08102, NIH SCORE GM08224, and NIH RISE GM61838.