Toxicological Research

  • 제12권1호
  • /
  • Pages.69-79
  • /
  • 1996
  • /
  • 1976-8257(pISSN)
  • /
  • 2234-2753(eISSN)
한국독성학회 (Korean Society of Toxicology & Korea Environmental Mutagen Society)
권호 표지

신경세포 배양법을 이용한 methamphetamine과 cadmium의 신경독성 평가

Neurotoxicity Assessment of Methamphetamine and Cadmium Using Cultured Neuronal Cells of Long-Evans Rats

  • 조대현 (식품의약품안전본부 독성연구소 독성부) ;
  • 김준규 (식품의약품안전본부 독성연구소 독성부) ;
  • 정용 (식품의약품안전본부 독성연구소 독성부) ;
  • 이봉훈 (식품의약품안전본부 독성연구소 독성부) ;
  • 김은엽 (식품의약품안전본부 독성연구소 독성부) ;
  • 김정구 (식품의약품안전본부 독성연구소 독성부) ;
  • 조태순 (성균관대학교 약학대학) ;
  • 김진석 (건국대학교 수의과대학) ;
  • 문화회 (식품의약품안전본부 독성연구소 독성부)
  • Cho, Dae-Hyun (Department of Toxicology, Korea Food and Drug Administration) ;
  • Kim, Jun-Gyon (Department of Toxicology, Korea Food and Drug Administration) ;
  • Jeong, Yong (Department of Toxicology, Korea Food and Drug Administration) ;
  • Lee, Bong-Hun (Department of Toxicology, Korea Food and Drug Administration) ;
  • Kim, Eun-Youb (Department of Toxicology, Korea Food and Drug Administration) ;
  • Kim, Jeong-Goo (Department of Toxicology, Korea Food and Drug Administration) ;
  • Cho, Tai-Soon (College of Pharmacy, Sung Kyun Kwan University) ;
  • Kim, Jin-Suk (College of Veterinary Medicine, Kon-Kuk University) ;
  • Moon, Hwa-Hwey (Department of Toxicology, Korea Food and Drug Administration)
  • 발행 : 1996.06.01
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초록

Primary culture of cerebellar neuronal cells derived from 8-day old Long-Evans rats was used. Pure granule cells, astrocytes or mixed cells culture systems were prepared. These cells were differentiated and developed synaptic connections. And the astrocytes were identified by immunostaining with glial fibrillary acidic protein (GFAP). Methamphetamine (MAP), which acts on dopaminergic system and cadmium (Cd), a toxic heavy metal, were applied and biochemical assays and electrophysiological studies were performed. $LC_50$ values estimated by MTT assay of MAP and Cd were 3 mM and 2$\mu M$ respectively. Cells were treated with 1 mM or 2 mM MAP and 1$\mu M$ $CdCl_2$ for 48 hour, and the incubation media were analyzed for the content of released LDH. MAP (2 mM) and Cd significantly increased the LDH release. Cell viability was decreased in both groups and some cytopathological changes like cell swelling or vacuolization were seen. The cerebellar granule cells were used for measuring membrane currents using whole-cell clamp technique. Sodium and potassium currents were not affected by MAP neither Cd, but calcium current was significantly reduced by Cd but not affected by MAP. Therefore, in vitro neurotoxicity test system using neuronaI cells and astrocytes cultures were established and can be used in screening of potential neurotoxic chemicals.

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참고문헌

  1. Neurotoxicology Neurotoxic agents Abou-Donia,M.B.
  2. Communication within animal cells Calcium Barritt,G.J.
  3. Toxicol. Lett. Methodology in neurotoxicityactivities within the World Health Organization and International Programme on Chemical Safety Becking,G.C.
  4. Patty's industrial hygiene and toxicology (4th ed.) v.3 Cadmium Beliles,R.P.;Clayton,G.D.(ed.);Clayton,F.E.(ed.)
  5. Neurotoxicol. Teratol. v.11 Intracellular calcium and neurotoxic events Bondy,S.C.
  6. Func. Neurol. v.8 Excitatory amino acids and neurotoxicity Bruno,V.;Scapagnini,U.;Canonico,P.L.
  7. Func. Neurol. v.9 Biomarkers in environmental medicine: alterations of cell signalling as early indicators of neurotoxicity Castoldi,A.F.;Coccini,T.;Rossi,A.;Nicotea,P.;Costa,L.G.;Tan,X.X.
  8. Intracellular messengers Calcium signalling and the triggering of secretion in adrenal chromaffin cells Cheek,T.R.;Taylor,C.W.(ed.)
  9. Neuron v.1 Glutamate neurotoxicity and disease of the nervous system Choi,D.W.
  10. Nature v.340 Rapid increase of an immediate early gene messenger RNA in hippocampal neurons by synaptic NMDA receptor activation Cole,A.J.;Saffen,D.W.;Barban,J.W.;Worley,P.F.
  11. Protocols for neural cell culture. Astrocytes and oilgodendrocyte culture Cole,R.;de Vellis,J.;Fedoroff,S.(ed.);Richardson,A.(ed.)
  12. Neurotoxicol. v.15 Signal transduction mechanisms in developmental neurotoxicity: the phosphoinositide pathway Costa,L.G.
  13. Incurabili. v.8 Sull'azione biologica del cadmio De Simone,G.
  14. Comp. Biochem. Phystol. C. v.100 Time-dependent cadmium-neurotoxicity in rat brain synaptosomal plasma membrance Fasitas,C.D.;Theocharis,S.E.;Zoulas,D.;Chrissimou,S.;Deliconstantinos,G.
  15. The cerebellum in Principles of Neural Science(3rd ed.) Ghez,C.;Kandel,E.R.(ed.);Schwartz,J.H.(ed.);Jessel,T.M.(ed.)
  16. Pfl$\ "{u}$ gers Arch. v.391 Improved patch-clamp techniques for high resolution current recording form cells and cell-free membrane patches Hamill,O.P.;Marty,A.;Neher,E.;Sakmann,B.;Sigworth,F.J.
  17. Anat. Rec. v.1 Observation on the living developing nerve fiber Harrison,R.
  18. J. Clin. Exp. Neuropsychol. v.11 Neuropsychological effects of occupational exposure to cadmium Hart,R.P.;Rose,C.S.;Hamer,R.M.
  19. J. Neurosci. v.8 Mouse cerebella granule neurons arrest the proliferation of human and rodent astroma cells in vitro Hatten,M.E.;Shelanski,M.L.
  20. Calcium channel in Ionic Channels of Excitable Membranes(2nd ed.) Hille,B.
  21. Arch. Toxicol. v.67 Neurons and glial cell marker proteins as indicators of heavy metal-induced neurotoxicity in neuroblastoma and glioma cell lines Huang,J.;Tanii,H.;Kato,K.;Hasimoto,K.
  22. Neurotoxicol. Teratol. v.13 Oxygen radicals: common mediators of neurotoxicity LeBel,C.P.
  23. Metab. Brain Dis. v.7 Differential effects of ions on type A and type B monoamine oxidase activities in rat brain and liver mitochondria. Leung,T.K.;Lim,L.;Lai,J.C.
  24. A dissection and Tissue Culture Manual of the Nervous System Preparation of 98% pure cerebellar granule cell culture. Levi,G.;Aloisi,F.;Ciotti,M.T.;Thangnipon,W.;Kinsbury,A.;Balazs,R.;Shahar,A.(ed.);de Vellis,J.(ed.);Vernadakis,A.(ed.);Haber,B.(ed.)
  25. Nature v.364 A redox-based mechanism for the neuroprotective and neurodegenerative effects of nitric oxide and related nitroso-com-punds Lipton,S.A.;Choi,Y.B.;Pan,Z.H.;Lei,S.Z.;Chen,H.S.;Sucher,N.J.;Loscalozo,J.;Single,D.J.;Stamler,J.S.
  26. Prog. Brain Res. v.103 Nitric oxide in the central nervous system Lipton,S.A.;Singel,D.J.;Stamler,J.S.
  27. J. Immuno. Meth. Rapid colorimetric assay for cellular growth and survival; Application to proliferation and cytotoxicity assays Mosmann,T.
  28. J. Trace Elem., Electrolytes Health Dis. v.7 Molecular mechanisms of metal neurotoxicity Nicotera,P.;Rossi,A.
  29. J. Neurochem. v.28 Cadmium alters behaviour and the biosynthetic capacity for catecholamines and serotonine in the neonatal rat brain Rastrogi,R.B.;Merali,Z.;Singhal,R.L.
  30. Acta Neuropathologica v.44 Cadmium-induced endothelial cell alterations in the fetal brain Rohrer,S.R.;Shaw,S.M.;Lamar,C.H.
  31. Neurotoxicol. v.15 Neurotoxicity and excitatory amino acid antagonists Schousboe,A.;Frandsen,A.;Whal,P.;Krogsgaard-Larsen,P.
  32. Br. J .Pharmacol. v.97 Inhibition of neurally-evoked transmitter release by calcium channel antagonists in rat parasympathetic ganglia Seabrook,G.R.;Adams,D.J.
  33. J. Cell Biol. v.45 Cerebellar granule cells in vitro. A light and electron microscope study Seil,F.J.;Herndon,R.M.
  34. FASEB J. v.6 Mechanisms of lead neurotoxicity, or looking beyond the lamppost Silbergeld,E.K.
  35. Neurotoxicol. Teratol. v.14 Modulation of glial cell differentiation by exposure to lead and cadmium Stark,M.;Wolff,J.E.;Korbmacher,A.
  36. Neurotoxicology The use of cell culture for evaluating neurotoxicity Veronesi,B.;Tilson,H.A.(ed.);Mitchell,C.L.(ed.)
  37. Drug Chem. Toxicol. v.14 Interaction of metals with brain calmodulin purified from normal and cadmium exposed rats Vig,P.J.;Ravi,K.;Nath,R.