Immunogold electron microscopic studies on stimulus-specific γ PKC translocation-a Synergistic effect of Ca^<2+> phorbol ester on PKC translocation
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概要
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We have studied the translocation of PKC subtypes in living cells using GFP-fusion protein to elucidate the function of individual PKC subtypes. We have demonstrated the subtype- and stimulus- specific targeting of PKC, suggesting that the targeting mechanism of PKC subtypes determines their individual role in cell signaling pathways. In the present study, we examined the precise intracellular translocation of the neuron-specific PKC subtype, γPKC (a postsynaptic PKC), after various extracellular stimuli using immunogold electron microscopy and also the synergistic effects of Ca^<2+> and phorbol ester on y PKC translocation. Under CLSM, both phorbol ester and Ca^<2+> inophone induced the translocation ofγPKC to the plasma membrane. Under electron microscopy, however, the precise localization of the translocated γPKC was differed after two stimulations.γPKC-GFP translocated onto the PM within 10 min. by the trearment of phorbol ester, TPA, while γPKC-GFP translocated to the periphery of the perikarya including PM within 20 sec. by the treatment of Ca^<2+>-ionophore A23187. Moreover, simultaneous treatment of these two stimuli revealed the rapid translocation of PKC to the same site as in the case with TPA treatment for more than 3 min. In conclusion, electron microscopic observation revealed that γPKC targets to different subcellular compartments in response to various stimuli and the synergistic effect suggested an additional mechanism of γPKC activation.
- 日本組織細胞化学会の論文
著者
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斎藤 尚亮
神戸大、バイオシグナル研究センター
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斉藤 尚亮
神大バイオシグナル
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斎藤 尚亮
神大バイオシグナル
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Saito Naoaki
Lab. Of Mol. Pharmacol. Biosignal Res. Ctr. Kobe University:grad Sch. Of Sci. & Tech. Kobe Unive
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FUJIMIYA Mineko
Department of Anatomy, Shiga University of Medical Science
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Fujimiya Mineko
Dept. Of Anatomy Shiga University Of Med. Sci.
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斎藤 尚亮
神戸大学バイオシグナル研究センタｰ分子薬理分野
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Saito N
Department Of Molecular And Pharmacological Neuroscience Graduate School Of Biomedical Sciences Hiro
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Oyasu Miho
Lab. of Mol. Pharmacol. Biosignal Res. Ctr., Kobe University
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Imaeda Hirotsugu
Dept. of Anatomy, Shiga University of Med. Sci.
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Fujimiya M
Department Of Anatomy Shiga University Of Medical Science
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Oyasu Miho
Lab. Of Mol. Pharmacol. Biosignal Res. Ctr. Kobe University:grad Sch. Of Sci. & Tech. Kobe Unive
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Imaeda Hirotsugu
Dept. Of Anatomy Shiga University Of Med. Sci.
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Sakai Norio
Department Of Molecular And Pharmacological Neuroscience Graduate School Of Biomedical Sciences Hiro
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