Toxicological Research

  • 제18권4호
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  • Pages.355-362
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  • 2002
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  • 1976-8257(pISSN)
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  • 2234-2753(eISSN)
한국독성학회 (Korean Society of Toxicology & Korea Environmental Mutagen Society)
권호 표지

Glutamate에 의한 세포내 칼슘농도변화와 세포독성과의 관계

Intracellular Calcium Concentration in the Glutamate-induced Cytotoxicity in PCl2 Cell

  • 발행 : 2002.12.01
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초록

Pathophysiological elevation of intracellular calcium concentration ($[Ca^{2+}]_1$) in the neuron has been considered as an important responsible factor in the neuronal cell damages. However the mechanism of increase of $[Ca^{2+}]_1$ and the relationship between $[Ca^{2+}]_1$ level and cytotocixity have not been fully demonstrated. In the present study, real-time alteration of $[Ca^{2+}]_1$and cellular response (cell damages) in the pheochromocytoma cells (PC12) stimulated by glutamate were investigated. Glutamate dose dependently decreased cell viability determined propidium iodide fluorescence method and morphology change. Conversely related with cell damages, glutamate dose dependently increased the level of［Ca$^{2+}$］$_{i}$ . To investigate the mechanism of glutamate-induced increase of $[Ca^{2+}]_1$,$[Ca^{2+}]_1$, was first measured in the cell cultured in calcium free media and in the presence of dantrolene, an inhibitor of calcium release from ryanodine receptor located in endoplasmic reticulum (ER). Similar to the increase$[Ca^{2+}]_1$ in the calcium-containing media, glutamate dose dependently increased $[Ca^{2+}]_1$ in the cell cultured in free calcium media. However pretreatment (2 hr) with 20~50 $\mu\textrm{M}$ dantrolene substantial lowered glutamate-induced increase of $[Ca^{2+}]_1$, suggesting that release of calcium from ER may be major sourse of increase of $[Ca^{2+}]_1$ in PC12 cells. Dantrolene-induced inhibition of $[Ca^{2+}]_1$ resulted in recovery of cytotoxicity by glutamate. Relevance of N-methy-D-aspartate (NMDA) receptor, a type of glutamte receptor on glutamate-induced incense of $[Ca^{2+}]_1$,$[Ca^{2+}]_1$ was also determined in the cells pretreated (2 hr) with NMDA receptor antagonist MK-80l. Glutamate-induced increase of $[Ca^{2+}]_1$ was reduced by MK-801 dose dependently. Furthermore, glutamate-induced cytotoxicity was also prevented by MK-80l. These results demonstrate that glutamte increase $[Ca^{2+}]_1$ dose dependently and thereby cause cytotoxicity. The increase of $[Ca^{2+}]_1$ may release from ER, especially through ryanodine receptor and/or through NMDA receptor Alteration of calcium homeostasis through disturbance of ER system and/or calcium influx through NMDA receptor could contribute glutamate-induced cell damages.s.

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참고문헌

  1. Soc. Neurosci. Abst. v.25 Raising intracellular calcium attenuates ischemia apoptosis in vitro Babcock,D.J.;Gottron,F.J.;Choi,D.W.
  2. Nature v.361 Inositol triphosphate and calcium signaling Berridge,M.J. https://doi.org/10.1038/361315a0
  3. Prog. Nucleic Acid Res. v.58 Regulation of translational initiation during cellular responsees to stress Brostrom,C.O.;Brostrom,M.A.
  4. Annu. Rev. Biochem. v.56 Intracellular calcium homeostasis Carafoli,E. https://doi.org/10.1146/annurev.bi.56.070187.002143
  5. J. Neurosci. v.10 Cerebral hypoxia:some new approaches and unanswered questions Choi,D.W.
  6. Trends Neurosci. v.18 Calcium:still center-stage in hypoxicischemic neuronal death Choi,D.W. https://doi.org/10.1016/0166-2236(95)93870-4
  7. Trends Neurosci. v.15 Suppression of programmed neuronal death by sustained elevation of cytoplasmic calcium Franklin,J.L.;Johnson,E.M.Jr. https://doi.org/10.1016/0166-2236(92)90103-F
  8. Brain Res. Mol. Brain Res. v.36 $Ca^{2+}$ entry following store depletion in SH-SY5Y neuroblastoma cells Grudt,T.J.;Usowicz,M.M.;Henderson,G. https://doi.org/10.1016/0169-328X(95)00248-Q
  9. J. Neurosci. v.17 Ionized intracellular cacium concentration predicts excitotoxic neuronal death:observations with low-affinity fluorescent cacium indicatiors Hyrc,K.;Handran,S.D.;Rothman,S.M.;Goldberg,M.P.
  10. Br. J. Pharmacol. v.112 Effects of bipyridylium compounds on calcium release fron triadic vesicles isolated from rabbit skeletal muscle Kang,J.J.;Hsu,K.S.;Lin Shiau,S.Y. https://doi.org/10.1111/j.1476-5381.1994.tb13213.x
  11. Stroke v.29 Calcium in ischemic cell death Kristian,T.Y.;Siesjo,B.Y. https://doi.org/10.1161/01.STR.29.3.705
  12. Trends Biochem. Sci. v.23 The endoplasmic reticulum $Ca^{2+}$ store:a view from the lumen Meldolesi,J.;Pozzan,T. https://doi.org/10.1016/S0968-0004(97)01143-2
  13. Biochem. J. v.327 Differential expression and regulation of ryanodine receptor and myoinositol 1,4,5-trisphosphate receptor $Ca^{2+}$ release channels in mammalian tissues and cell lines Mackrill,J.J.;Challiss,R.A.J.;O'connell,D.A.;Lai,F.A.;Nahorski,S.R. https://doi.org/10.1042/bj3270251
  14. J. Neurochem. v.72 Alterations of extracellular calcium elicit selective modes of cell death and protease activation in SH-SY5Y human neuroblastoma cells MaGinnis,K.M.;Wang,K.K.;Gnegy,M.E. https://doi.org/10.1046/j.1471-4159.1999.0721853.x
  15. Biochem. Mol. Biol. Int. v.43 Thapsigargin induces apoptosis in SH-SY5Y neuroblastoma cells and cerebrocortical cultures Nath,R.;Raser,K.J.;Hajimohammadreza,I.;Wang,K.K.
  16. J. Neural Transm. Suppl. v.43 The calcium ion and cell death Orrenius,S.;Nicotera,P.
  17. J. Cereb. Blood Flow Metab. v.19 Disturbances of the functioning of endoplasmic reticulum:a key mechanism underlying neuronal cell injury? Paschen,W.;Doutheil,J. https://doi.org/10.1038/sj.bmt.1700655
  18. Trends Cell. Biol. v.7 Endoplasmic-reticulum-induced signal transduction and gene expression Pahl,F.L.;Baeuerle,P.A. https://doi.org/10.1016/S0962-8924(96)10050-7
  19. Physiol. Rev. v.79 Signal transduction fron the endoplasmic reticulum to the cell nucleus Pahl,F.L. https://doi.org/10.1152/physrev.1999.79.3.683
  20. J. Neurochem. v.67 Depletion of neuronal endoplasmic reticulum calcium stores by thapsigagin:effect on protein synthesis Paschen,W.;Doutheil,J.;Gissel,C.;Treiman,M. https://doi.org/10.1046/j.1471-4159.1996.67041735.x
  21. Physiol. Rev. v.74 Molecular and cellular physiology of intracellular calcium stores Pozzan,T.;Rizzito,R.;Volpe,P.;Meldolesi,J. https://doi.org/10.2466/pr0.1994.74.2.595
  22. Cancer Res. v.51 Differential expression of bcl-2 protooncogene in neuroblastoma and other human tumor cell lines of neural origin Reed,J.C.;Meister,L.;Tanaka,S.;Cuddy,M.;Yum,S.;Geyer,C.;Pleasure,D.
  23. J. Neurochem. v.71 Distinct influx pathways, not calcium load, determine neuronal vulnerability to calcium neurotoxicity Sattler,R.;Charlton,M.P.;Hafner,M.;Tymianski,M. https://doi.org/10.1046/j.1471-4159.1998.71062349.x
  24. J. Cereb. Blood Flow Metab. v.9 Calcium fluxes, calcium antagonists, and calcium-related pathology in brain ischemia, hypoglycemia, and spreading depression:a unifying hypothesis Siesjo,B.K.;Bengtsson,F. https://doi.org/10.1038/jcbfm.1989.20
  25. J. Cereb. Blood Flow Metab. v.19 Is the cell death pathway triggered by the mitochondrion or the endoplasmic reticulum? Siesjo,B.K.;Hu,B.;Kristian,T. https://doi.org/10.1097/00004647-199901000-00002
  26. Neurosurgery v.38 Normal and abnormal calcium homeostasis in neurons:a basis for the pathophysiology of traumatic and ischemic central nervous system injury Tymianski,M.;Tator,C.H. https://doi.org/10.1097/00006123-199606000-00028
  27. Drugs v.32 Dantrolene. A review of its pharmacodynamic and pharmacokinetic properties and therapeutic use in malignant hyperthermia, the neuroleptic malignant syndrome and an update of its use in muscle spasticity Ward,A.;Chaffman,M.O.;Sorkin,E.M. https://doi.org/10.2165/00003495-198632020-00003
  28. J. Neurochem. v.67 Dantrolene is cytoprotective in two medels of neuronal cell death Wei,H.;Perry,D.C. https://doi.org/10.1046/j.1471-4159.1996.67062390.x
  29. J. Neurochem. v.70 Bcl-2 protects against apoptosis in neurnal cell line caused by thapsogargin-induced depletion of intracellular calcium stores Wei,H.;Wei,W.;Dredesen,D.E.;Perry,D.C. https://doi.org/10.1046/j.1471-4159.1998.70062305.x