Effect of Methanol on Cultured Neuronal and Glial Cells on Rat Hippocampus

Methanol이 배양된 흰쥐 해마의 신경세포 및 신경교 세포의 성장에 미치는 영향

  • 이정임 (이안과 의원) ;
  • 조병채 (이화여자대학교 의과대학 안과학실) ;
  • 배영숙 (이화여자대학교 의과대학 약리학실) ;
  • 이경은 (이화여자대학교 의과대학 약리학실)
  • Published : 1996.12.01

Abstract

Methanol has been widely used as an industrial solvent and environmental exposure to methanol would be expected to be increasing. In humans, methanol causes metabolic acidosis and damage to ocular system, and can lead to death in severe and untreated case. Clinical symptoms are attributed to accumulation of forrnic acid which is a metabolic product of methanol. In humans and primates, formic acid is accumulated after methanol intake but not in rodents due to the rapid metabolism of methanol. Neverthless, the developmental and reproductive toxicity were reported in rodents. Previous reports showed that perinatal exposure to ethanol produces a variety of damage in human central nervous system by direct neurotoxicity. This suggests that the mechanism of toxic symptoms by methanol in rodents might mimic that of ethanol in human. In the present study I hypothesized that methanol can also induce toxicity in neuronal cells. For the study, primary culture of rat hippocampal neurons and glias were empolyed. Hippocampal cells were prepared from the embryonic day-17 fetuses and maintained up to 7 days. Effect of methanol (10, 100, 500 and 1000 mM) on neurite outgrowth and cell viability was investigated at 0, 18 and 24 hours following methanol treatment. To study the changes in proliferation of glial cells, protein content was measured at 7 days. Neuronal cell viability in culture was not altered during 0-24 hours after methanol treatment. 10 and 100 mM methanol treatment significantly enhanced neurite outgrowth between 18-24 hours. 7-day exposure to 10 or 100 mM methanol significantly increased protein contents but that to 1000 mM methanol decreased in culture. In conclusion, methanol may have a variety of effects on growing and differentiation of neurons and glial cells in hippocampus. Treatment with low concentration of methanol caused that neurite outgrowth was enhanced during 18-24 hours and the numbers of glial cell were increased for 7 days. High concentration of methanol brought about decreased protein contents. At present, the mechanism responsible for the methanol- induced enhancement of neurite outgrowth is not clear. Further studies are required to delineate the mechanism possibly by employing molecular biological techniques.

Keywords

References

  1. Pharmacol. Biochem. Behav. v.10 Prenatal effects of alcohol on adult learning in rats Abel,E.L.
  2. Teratol. v.49 Effects of methanol on embryonic mouse palate in serum-free organ culture Abbott,B.D.;Logsdon,T.R.;Wilke,T.S.
  3. J. Comp. Neurol. v.347 Differential expression of five N-methyl-D-aspartate receptor subunit mRNAs in the cerebellum of developing and adult rats Akazawa,C.;Shigemoto,R.;Bessho,Y.;Nakanishi,S.;Mizuno,N.
  4. Anal. Biochem. v.72 A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Badford,M.
  5. Neuroreport v.5 no.8 Expression of the NMDA R1 receptor in selected human brain regions Bockers,T.M.;Zimmer,M.;Muller,A.;Bergmann,M.;Brose,N.;Kreutz,M.R.
  6. Alcohol Clin. Exp. Res. v.14 Alcohol-induced neuronal loss in developing rats:increased brain damage with binge exposure Bonthius,D.J.;West,J.R.
  7. Neurobehav. Toxicol. Teratol. v.5 Effects of gestational alcohol on the development of neon atal reflexes in the rat Cogan,D.C.;Cohen,L.E.;Sparman,G.
  8. J. Neurosci. v.7 Electrically and chemically mediated increases in intracellular calcium in neuronal growth cones Cogan,C.S.;Conner,J.A.;Kater,S.B.
  9. Proc. Natl. Acad. Sci. USA v.83 Digital imaging of free calcium changes and of spatial gradients in growing processes in single mammalian central nervous system cells Connor,J.A.
  10. Science v.208 Impaired brain growth in neonatal rats exposed to ethanol Diaz,J.;Samson,H.H.
  11. Alcohol v.9 Ethanol and voltage - or receptor - mediatd increases in cytosolic Ca in brain cells Dildi Mayfield,J.E.;Machu,T.;Leslie,S.W.
  12. Nature v.368 NMDA-receptor channel diversity in the developing cerebellum Farrant,M.;Feldmeyer,D.;Takahashi,T.;Cull Candy,S.G.
  13. Neurobehav. Toxicol. Teratol. v.5 Effects of prenatal alcohol on homong behavior, maternal responding and openfield activity in rats Fernandez,K.;Caul,W.P.;Haenlein,M.;Vorhees,C.V.
  14. Am. J. Ophthalmol. v.39 Studies on visual toxicity of methanol;role of acidosis in experimental methanol poisoning Gilger,A.P.;Potts,A.M.
  15. Am. J. Ophthalmol. v.42 Studies on visual toxicity of methanol;effects of ethanol on methanol poisoning in rhesus monkey Gilger,A.P.;Potts,A.M.;Farkas,I.S.
  16. Proc. Natl. Acad. Sci. USA v.83 Protein kinase C as a component of a nerve growth factor-sensitive phosphorylation system in PC12 cells Hama,T.;Huang,K.P.;Guroff,G.
  17. J. Histochem. Cytochem. v.33 An improved method to determine cell viability by simultaneous staining wity fluorescein diacetate propedium iodide Jones,K.H.;Senft,J.A.
  18. J. Neurosci. v.11 Regulation of growth cone behavior by calcium Kater,S.B.;Mills,L.R.
  19. Int. J. Develop. Neurosci. v.6 Isolated hippocampal neurons in cryopreserved long-term cultures:Development of neruoarchitecture and sensitivity to NMDA Mattson,M.P.;Kater,S.V.
  20. J. Neurosci. v.7 Calcium regulatation of neurite elongation and growth cone motility Mattson,M.P,;Katter,S.B.
  21. Neurosci. Biobehav. Rev. v.2 The effects of protein malnutrition on the developing central nervous system in rat Morgane,P.J.;Miller,M.W.;Kemper,T.;Stern,W.;Forbes, W.;Hall,R.;Bronzino,J.;Kissane,J.;Hawrylewicz,E.;Resnick,O.
  22. Science v.222 Modulation of synapse formation by cyclic adenosine monophosphate Nirenberg,M.;Wilson,S.;Higashida,H.;Rotter,A.;Krueger,K.;Busis,N.;Ray,R.;Kenimer,J.G.;Adler,M.
  23. Develop. Brain Res. v.5 Electrophysiological effects of acute ethanol exposure Ⅱ. Alterations in the calcium component of action potentials from sensory neurons in dissociated culture Oakes,S.G.;Pozos,R.S.
  24. Trends Neurosci. v.6 Undernutrition and brain development Patel,A.J.
  25. Alcohol v.9 Alcohol reduces the number of pheochromocytoma (PC12) cells in culture Pantazis N.J.;Dohrman,D.P.;Luo,J.;Goodlett,C.R.;West,J.R.
  26. Am. J. Ophthalmol. v.39 Visual toxicity of methanol:clinical aspects of experimental methanol poisoning treated with base Potts,A.M.
  27. Am. J. Ophthalmol. v.35 Studies on visual toxicity of methanol;efects of methanol and its degradation products on retinal metabolism Potts,A.M.;Johnson,L.V.
  28. Am. J. Ophthalmol. v.40 Studies on visual toxicity of methanol;additional observations on methanol poisoning in primate test object Potts,A.M.;Praglin,J.;Farkas,I.S.;Orbison,L.;Chickering,D.
  29. CRC Critt. Rev. Toxicol. v.10 Species differences in methanol poisoning ReO.
  30. Alcohol Clin. Exp. Res. v.10 Effects of ethanol exposure during the third trimester equivalent on neuron number in rat hippocampus and dentate gyrus West,J.R.;Hamre,K.M.;Cassell,M.D.
  31. Brain Res. v.550 Alcohols synergize with NGF to induce earlly differention of PC12 cells Wooten,M.W.;Edward,S.J.
  32. Develop. Brain Res. v.72 Ethanol enhances neurite outgrowth in primary cultures of rat cerebellar macroneurons Zou,Z.Y.;Rabin,R.A.;Pentney,R.J.
  33. 대한의학협회지 v.1 알콜의 약리 홍사석