Abstract
An antisense (AS) oligodeoxynucleotide based on a conserved sequence in the three isoforms of the Na(+)/Ca(2+) exchanger (NCX) was used to decrease expression of this Ca(2+) transporter in primary neuronal cultures. Two AS oligo applications decreased NCX activity by approximately 40% within 12-24 h, and neither sense (S) or missense (MS) oligos altered NCX activity. The reduced NCX expression was confirmed by immunoblots and enzyme-linked immunosorbent assays (ELISAs). Resting [Ca(2+)](i) levels were 20% higher in AS-treated neurons and showed a slower return to baseline levels following activation of Ca(2+) influx by N-methyl-D-aspartate (NMDA). These results suggest that NCX plays a significant role in maintaining neuronal Ca(2+) homeostasis and in restoring baseline Ca(2+) levels following depolarization.
Publication types
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Animals
-
Brain / cytology
-
Brain / drug effects*
-
Brain / metabolism
-
Calcium / metabolism
-
Cells, Cultured
-
Conserved Sequence / genetics
-
Enzyme-Linked Immunosorbent Assay
-
Fluorescent Dyes
-
Fura-2
-
Homeostasis / drug effects
-
Immunoblotting
-
Intracellular Fluid / metabolism
-
N-Methylaspartate / metabolism
-
N-Methylaspartate / pharmacology
-
Neurons / cytology
-
Neurons / drug effects*
-
Neurons / metabolism
-
Oligonucleotides, Antisense / genetics
-
Oligonucleotides, Antisense / pharmacology*
-
Protein Isoforms / antagonists & inhibitors
-
Protein Isoforms / genetics
-
Rats
-
Rats, Sprague-Dawley
-
Sodium-Calcium Exchanger / antagonists & inhibitors*
-
Sodium-Calcium Exchanger / genetics
-
Sodium-Calcium Exchanger / metabolism
Substances
-
Fluorescent Dyes
-
Oligonucleotides, Antisense
-
Protein Isoforms
-
Sodium-Calcium Exchanger
-
N-Methylaspartate
-
Calcium
-
Fura-2