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Molecular Pharmacology

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Research ArticleArticle

Overexpression of Glyceraldehyde-3-Phosphate Dehydrogenase Is Involved in Low K+-Induced Apoptosis but Not Necrosis of Cultured Cerebellar Granule Cells

Ryoichi Ishitani, Katsuyoshi Sunaga, Masaharu Tanaka, Hideki Aishita and De-Maw Chuang
Molecular Pharmacology April 1997, 51 (4) 542-550; DOI: https://doi.org/10.1124/mol.51.4.542
Ryoichi Ishitani
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Katsuyoshi Sunaga
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Masaharu Tanaka
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Hideki Aishita
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De-Maw Chuang
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Abstract

We have reported that overexpression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) is involved in age-induced apoptosis of the cultured cerebellar granule cells that grow in a depolarizing concentration (25 mm) of KCl. The present study was undertaken to investigate whether GAPDH overexpression also occurs and participates in apoptosis of the cerebellar granule cells that result from switching the culturing conditions from high (25 mm) to low (5 mm) concentrations of KCl. We found that exposure of granule cells to low potassium (K+) for 24 hr induces not only apoptosis but also necrotic damage. The latter is supported by the morphological observations that a subpopulation of neurons showed cell swelling, extensive cytoplasmic vacuolization, damaged mitochondria, and apparently intact nuclei. Treatments with two antisense but not sense oligodeoxyribonucleotides directed against GAPDH attenuated low K+-induced neuronal death by approximately 50%. Morphological inspection revealed that GAPDH antisense oligonucleotides preferentially blocked low K+-induced apoptosis with little or no effect on necrotic damage. Similar to antisense oligonucleotides, actinomycin-D partially inhibited low K+-induced death of granule cells with a predominant effect on apoptosis. In contrast, cycloheximide almost completely blocked low K+-induced neuronal death and seemed to prevent both apoptotic and necrotic damage. The levels of GAPDH mRNA and protein were markedly increased in a time-dependent manner after low K+ exposure. The overexpression of GAPDH mRNA and protein was completely blocked by cycloheximide, actinomycin-D, and its antisense but not sense oligonucleotides. Taken together, these results lend credence to the view that exposure of cerebellar granule cells to low K+ induces both apoptosis and necrosis and that only the apoptotic component involves overexpression of GAPDH.

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Molecular Pharmacology: 51 (4)
Molecular Pharmacology
Vol. 51, Issue 4
1 Apr 1997
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Research ArticleArticle

Overexpression of Glyceraldehyde-3-Phosphate Dehydrogenase Is Involved in Low K+-Induced Apoptosis but Not Necrosis of Cultured Cerebellar Granule Cells

Ryoichi Ishitani, Katsuyoshi Sunaga, Masaharu Tanaka, Hideki Aishita and De-Maw Chuang
Molecular Pharmacology April 1, 1997, 51 (4) 542-550; DOI: https://doi.org/10.1124/mol.51.4.542

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Research ArticleArticle

Overexpression of Glyceraldehyde-3-Phosphate Dehydrogenase Is Involved in Low K+-Induced Apoptosis but Not Necrosis of Cultured Cerebellar Granule Cells

Ryoichi Ishitani, Katsuyoshi Sunaga, Masaharu Tanaka, Hideki Aishita and De-Maw Chuang
Molecular Pharmacology April 1, 1997, 51 (4) 542-550; DOI: https://doi.org/10.1124/mol.51.4.542
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