ADENOVIRUS-MEDIATED EXPRESSION OF AMPA-TYPE GLUTAMATE-RECEPTOR CHANNELS IN PC12 CELLS

Citation
M. Sudo et al., ADENOVIRUS-MEDIATED EXPRESSION OF AMPA-TYPE GLUTAMATE-RECEPTOR CHANNELS IN PC12 CELLS, Molecular brain research, 50(1-2), 1997, pp. 91-99
Citations number
36
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Neurosciences
Journal title
ISSN journal
0169-328X
Volume
50
Issue
1-2
Year of publication
1997
Pages
91 - 99
Database
ISI
SICI code
0169-328X(1997)50:1-2<91:AEOAGC>2.0.ZU;2-7
Abstract
The lpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid(AMPA)-type glutamate receptor channels are expressed ubiquitously in brain neuro ns and mediate fast excitatory neurotransmission. They are composed of four subunits, GluR1, GluR2, GluR3 and/or GluR4. We constructed recom binant adenoviruses encoding rat AMPA receptor subunit cDNAs, GluR1 (A xCAGluR1) and silently mutated GluR2 (AxCAGluR2X) with modified chicke n beta-actin promoter and cytomegalovirus immediate-early enhancer. Us ing these adenoviral vectors, we transferred the GluR1 and GluR2 genes into PC12 cells that possess no functional AMPA receptor channels. PC 12 cells infected with these viruses expressed GluR1 and GluR2 RNAs. I mmunoblot analysis indicated that the expressed GluR1 and GluR2 protei ns were equivalent to those of the rat brain. Functional expression of the AMPA receptor channels was examined using the whole-cell patch cl amp technique. In AxCAGluR1-infected cells, the current-voltage (I-V) relationship of response to kainate, a non-desensitizing agonist of AM PA receptors, exhibited a strong inward rectification, indicating the formation of functional GluR1-homomeric channels. In cells infected wi th both AxCAGluR1 and AxCAGluR2X, the I-V relationship of kainate resp onses exhibited an outward rectification, indicating the formation of heteromeric GluR1/R2 channels. Immunocytochemical analysis revealed th at the AMPA receptor subunit genes were transferred in more than 95% o f the infected PC12 cells. (C) 1997 Elsevier Science B.V.