Cell-permeable cAMP analog suppresses 6-hydroxydopamine-induced apoptosis in PC12 cells through the activation of the Akt pathway

Hirofumi Fujita, Tetsuya Ogino, Hirotsugu Kobuchi, Takuzo Fujiwara, Hiromi Yano, Jitsuo Akiyama, Kozo Utsumi, Junzo Sasaki

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Although cAMP protects neuronal cells from various apoptotic stimulations, its mechanism is not fully elucidated. We report here the molecular mechanism of the 6-hydroxydopamine (6-OHDA)-induced apoptosis of pheochromocytoma PC12 cells and its suppression by 8-(4-chlorophenylthio)adenosine 3′,5′-cyclic monophosphate (pCPT-cAMP), which is a membrane permeable cAMP analog. Treatment of PC12 cells with 6-OHDA resulted in the activation of caspases and apoptosis, as detected by chromatin condensation. 6-OHDA also induced superoxide generation, Bid cleavage and mitochondrial membrane depolarization. In addition, Akt phosphorylation that was favorable to cell survival was decreased and p38 MAPK phosphorylation was increased by 6-OHDA. PC12 cell apoptosis was inhibited by pCPT-cAMP, Z-VAD-fmk (a broad-range caspase inhibitor) and tiron (a superoxide scavenger), although PC12 cell apoptosis was not inhibited by cyclosporine A (an inhibitor of mitochondrial membrane permeability transition). Moreover, pCPT-cAMP promoted Akt phosphorylation, but it did not prevent superoxide generation and mitochondrial membrane depolarization. Conversely, LY294002, an inhibitor of Akt upstream molecule PI3-kinase, enhanced 6-OHDA-induced apoptosis. These results indicated that the 6-OHDA-induced apoptosis of PC12 cells was initiated by superoxide generation followed by caspase cascade activation, which was associated with the suppressed Akt phosphorylation and increased p38 phosphorylation. It is likely that pCPT-cAMP prevented the 6-OHDA-induced apoptosis via activation of the PI3-kinase/Akt pathway without any effect on superoxide generation or mitochondrial membrane depolarization.

Original languageEnglish
Pages (from-to)10-23
Number of pages14
JournalBrain Research
Volume1113
Issue number1
DOIs
Publication statusPublished - Oct 3 2006

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Oxidopamine
PC12 Cells
Apoptosis
Superoxides
Mitochondrial Membranes
Phosphorylation
Caspases
Phosphatidylinositol 3-Kinases
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Caspase Inhibitors
Pheochromocytoma
p38 Mitogen-Activated Protein Kinases
Adenosine
Cyclosporine
Chromatin
Permeability
Cell Survival
Membranes

Keywords

  • 6-Hydroxydopamine
  • Akt
  • Apoptosis
  • cAMP
  • Mitochondrial membrane potential
  • PC12 cell

ASJC Scopus subject areas

  • Neuroscience(all)
  • Clinical Neurology
  • Developmental Biology
  • Molecular Biology

Cite this

Cell-permeable cAMP analog suppresses 6-hydroxydopamine-induced apoptosis in PC12 cells through the activation of the Akt pathway. / Fujita, Hirofumi; Ogino, Tetsuya; Kobuchi, Hirotsugu; Fujiwara, Takuzo; Yano, Hiromi; Akiyama, Jitsuo; Utsumi, Kozo; Sasaki, Junzo.

In: Brain Research, Vol. 1113, No. 1, 03.10.2006, p. 10-23.

Research output: Contribution to journalArticle

Fujita, Hirofumi ; Ogino, Tetsuya ; Kobuchi, Hirotsugu ; Fujiwara, Takuzo ; Yano, Hiromi ; Akiyama, Jitsuo ; Utsumi, Kozo ; Sasaki, Junzo. / Cell-permeable cAMP analog suppresses 6-hydroxydopamine-induced apoptosis in PC12 cells through the activation of the Akt pathway. In: Brain Research. 2006 ; Vol. 1113, No. 1. pp. 10-23.
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AU - Fujiwara, Takuzo

AU - Yano, Hiromi

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AB - Although cAMP protects neuronal cells from various apoptotic stimulations, its mechanism is not fully elucidated. We report here the molecular mechanism of the 6-hydroxydopamine (6-OHDA)-induced apoptosis of pheochromocytoma PC12 cells and its suppression by 8-(4-chlorophenylthio)adenosine 3′,5′-cyclic monophosphate (pCPT-cAMP), which is a membrane permeable cAMP analog. Treatment of PC12 cells with 6-OHDA resulted in the activation of caspases and apoptosis, as detected by chromatin condensation. 6-OHDA also induced superoxide generation, Bid cleavage and mitochondrial membrane depolarization. In addition, Akt phosphorylation that was favorable to cell survival was decreased and p38 MAPK phosphorylation was increased by 6-OHDA. PC12 cell apoptosis was inhibited by pCPT-cAMP, Z-VAD-fmk (a broad-range caspase inhibitor) and tiron (a superoxide scavenger), although PC12 cell apoptosis was not inhibited by cyclosporine A (an inhibitor of mitochondrial membrane permeability transition). Moreover, pCPT-cAMP promoted Akt phosphorylation, but it did not prevent superoxide generation and mitochondrial membrane depolarization. Conversely, LY294002, an inhibitor of Akt upstream molecule PI3-kinase, enhanced 6-OHDA-induced apoptosis. These results indicated that the 6-OHDA-induced apoptosis of PC12 cells was initiated by superoxide generation followed by caspase cascade activation, which was associated with the suppressed Akt phosphorylation and increased p38 phosphorylation. It is likely that pCPT-cAMP prevented the 6-OHDA-induced apoptosis via activation of the PI3-kinase/Akt pathway without any effect on superoxide generation or mitochondrial membrane depolarization.

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KW - Mitochondrial membrane potential

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