Antisense oligonucleotide suppression of Na(+)/Ca(2+) exchanger activity in primary neurons from rat brain.

Ranciat-McComb, N S; Bland, K S; Huschenbett, J; Ramonda, L; Bechtel, M; Zaidi, A; Michaelis, M L

Ranciat-McComb, N S; Bland, K S; Huschenbett, J; Ramonda, L; Bechtel, M; Zaidi, A; Michaelis, M L.

Afiliación

Ranciat-McComb NS; Department of Pharmacology and Toxicology, 5064 Malott Hall, and Center for Neurobiology and Immunology Research, University of Kansas, Lawrence, KS 66045, USA.

Neurosci Lett ; 294(1): 13-6, 2000 Nov 10.

Article en En | MEDLINE | ID: mdl-11044575

RESUMEN

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.

Asunto(s)

Encéfalo/efectos de los fármacos; Neuronas/efectos de los fármacos; Oligonucleótidos Antisentido/farmacología; Intercambiador de Sodio-Calcio/antagonistas & inhibidores; Animales; Encéfalo/citología; Encéfalo/metabolismo; Calcio/metabolismo; Células Cultivadas; Secuencia Conservada/genética; Ensayo de Inmunoadsorción Enzimática; Colorantes Fluorescentes; Fura-2; Homeostasis/efectos de los fármacos; Immunoblotting; Líquido Intracelular/metabolismo; N-Metilaspartato/metabolismo; N-Metilaspartato/farmacología; Neuronas/citología; Neuronas/metabolismo; Oligonucleótidos Antisentido/genética; Isoformas de Proteínas/antagonistas & inhibidores; Isoformas de Proteínas/genética; Ratas; Ratas Sprague-Dawley; Intercambiador de Sodio-Calcio/genética; Intercambiador de Sodio-Calcio/metabolismo

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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Encéfalo / Oligonucleótidos Antisentido / Intercambiador de Sodio-Calcio / Neuronas Límite: Animals Idioma: En Revista: Neurosci Lett Año: 2000 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Irlanda

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