Antisense oligonucleotide suppression of Na(+)/Ca(2+) exchanger activity in primary neurons from rat brain.
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.
Ranciat-McComb
N S
NS
Department of Pharmacology and Toxicology, 5064 Malott Hall, and Center for Neurobiology and Immunology Research, University of Kansas, Lawrence, KS 66045, USA.
Bland
K S
KS
Huschenbett
J
J
Ramonda
L
L
Bechtel
M
M
Zaidi
A
A
Michaelis
M L
ML
D000818:Animals; D001921:Brain / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D017124:Conserved Sequence / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes; D016257:Fura-2; D006706:Homeostasis / Q000187:drug effects; D015151:Immunoblotting; D007424:Intracellular Fluid / Q000378:metabolism; D016202:N-Methylaspartate / Q000378:metabolism / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D019831:Sodium-Calcium Exchanger / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
D005456:Fluorescent Dyes; D016376:Oligonucleotides, Antisense; D020033:Protein Isoforms; D019831:Sodium-Calcium Exchanger; D016202:N-Methylaspartate; D002118:Calcium; D016257:Fura-2