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Differential changes in cell morphology, macromolecular composition and membrane protein profiles of neurons and astrocytes in chronic ethanol treated rats

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Abstract

Cellular morphology, macromolecular composition, (DNA, RNA and Protein content) marker enzyme activities for neurons [neuron specific enolase (NSE)] and astrocytes [glutamine synthetase (GS)] and plasma membrane protein profiles in the bulk isolated neurons and astrocytes from control and ethanol treated rats were studied. One month aged Wistar rats were given ethanol as sole drinking fluid for 10 weeks. Scanning electron microscopy revealed a characteristic cell surface smoothening in astrocytes due to ethanol treatment. DNA levels were unaltered, while RNA and Protein contents were decreased in astrocytes and neurons. Further,3H-leucine incorporation into proteins was decreased in neurons and astrocytes derived from ethanol treated rats indicating reduced protein synthesis in neurons and astrocytes. GS activity was affected severely suggesting impairment in astrocytic functions. Plasma membrane protein composition was analyzed by 2-D electrophoresis. The analysis indicated several protein defects in the plasma membranes of neurons and astrocytes, which might be involved in ‘membrane disorder’ during ethanol challenge.125I-Wheat Germ agglutinin binding studies showed three prominent proteins (160, 116 and 97 kDa) in astrocyte membrane fraction suggesting the possible involvement of N-terminal glycoproteins in altered astrocyte morphology during ethanol ingestion. Impairment in the astrocyte cell functions, protein changes in plasma membrane and cellular morphology studies suggest that astrocytes may be more vulnerable than neurons for ethanol effects.

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References

  1. Hamsson T, Tindberg N, Ingelman-Sundberg M, Kohler C: Regional distribution of ethanol inducible cytochrome P450 IIE1 in the rat central nervous system. Neuroscience 34: 451–463, 1990

    PubMed  Google Scholar 

  2. Kotch EL, Sulik KK: Patterns of ethanol induced cell death in the developing nervous system of mice: Neural fold states through the time of anterior neural tube closure. Int J Dev Neuroscience 10: 273–279, 1992

    Google Scholar 

  3. West JR, Goodlett CR: Teratogenic effects of alcohol on brain development. Ann Med 22: 319–325, 1990

    PubMed  Google Scholar 

  4. Peiffer J, Majewski F, Fischbach F, Bierich JR, Volk B: Alcohol embryo and fetopathy. J Neurol Sci 41: 125–137, 1979

    PubMed  Google Scholar 

  5. Phillips SC, Cragg BG: A change in susceptibility of rat cerebellar Purkinje cells to damage by alcohol during fetal, neonatal and adult life. Neuropathol Appl Neurobiol 8: 441–454, 1982

    PubMed  Google Scholar 

  6. Fleming EW, Woodson ME, Tewari S: Ethanol and cycloheximide alter protein and RNA synthesis of Cox astrocytoma cells in culture. J Neurosci Res 6: 511–524, 1981

    PubMed  Google Scholar 

  7. Kiianmaa K: Neuronal mechanisms of ethanol sensitivity. Alcohol & Alcoholism 25: 257–262, 1990

    Google Scholar 

  8. Davies DL, Vernadakis A: Effects of ethanol on cultured glial cells: Proliferation and glutamine synthetase activity. Dev Brain Res 16: 27–35, 1984

    Google Scholar 

  9. Guerri G, Saez R, Sancho-Tello M, Martinde Aquilera E, Renau-Piqueras J: Ethanol alters astrocyte development: A study of critical periods using primary cultures. Neurochemical Res 15: 559–565, 1990

    Google Scholar 

  10. Massarelli R, Syapin PJ, Noble EP: Increased uptake of choline by neural cell cultures chronically exposed to ethanol. Life Sci 18: 397–404, 1976

    PubMed  Google Scholar 

  11. Noble EP, Syapin PJ, Vigran R, Rosenberg A: Neuroaminidase releasable surface sialic acid of cultured astroblasts exposed to ethanol. J Neurochem 27: 217–221, 1976

    PubMed  Google Scholar 

  12. Fuches V, Coper H, Romelspacher H: The effect of ethanol and haloperiodo on dopamine receptor (D2) density. Neuropharmacology 26: 1231–1233, 1987

    PubMed  Google Scholar 

  13. Babu PP, Nagaraju N, Vemuri MC: Differences in the plasma membrane proteins of chronic alcoholic rat brain. Membrane Biochem 9: 227–237, 1990

    PubMed  Google Scholar 

  14. Haviryaji KSG, Srinivas R, Suryanarayana T, Vemuri MC: Effect of ethanol on hepatic Ribosomal proteins and mRNA. Mol Cell Biochem 115: 143–147, 1992

    PubMed  Google Scholar 

  15. Farooq M, Norton WJ: A modified procedure for isolation of astrocyte and neuron enriched fractions from rat brain. J Neurochem 31: 887–894, 1978

    PubMed  Google Scholar 

  16. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ: Protein measurement with folin phenol reagent. J Biol Chem 193: 265–275, 1951

    PubMed  Google Scholar 

  17. Schmidt G, Thannhansen SJ: In: G Keleti, WH Lederer (eds.) Handbook of Micro Methods for the Biological Sciences. Van Nostrand Reinhold Company, 1974, pp 9–10

  18. Bergmeyer HU, Bernt E: In: HU Bergmeyer (ed.) Methods of Enzymatic Analysis. Vol. I. Academic Press, NY, 1974, pp 449

    Google Scholar 

  19. Rowe WB, Ronzio RA, Welner VP, Meister A: Glutamine synthetase (Sheep brain). Methods Enzymol XVIIA, 1970, pp 900–911

    Google Scholar 

  20. Pinkett O, Anderson WB: Alterations in plasma membrane proteins associated with the proteolytic activation of adenylate cyclase. Arch Biochem Biophys 200: 261–268, 1980

    PubMed  Google Scholar 

  21. O'Farrel PH: High resolution two dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021, 1975

    PubMed  Google Scholar 

  22. Blum H, Beier H, Gross HJ: Improved silver staining of plant proteins, RNA and DNA polyacrylamide gels. Electrophoresis 8: 93–99, 1987

    Google Scholar 

  23. Glass II WF, Briggs RC, Hnilica LS: Use of lectins for detection of electrophoretically separated glycoproteins transferred on to nitrocellulose sheets. Anal Biochem 115: 210–224, 1981

    Google Scholar 

  24. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  25. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354, 1979

    PubMed  Google Scholar 

  26. Klemm WR, Engen RL: Biochemical markers of ethanol effects on brain. J Neuro Sci Res 3: 341–351, 1978

    Google Scholar 

  27. Kennedy LA, Mukerji S: Ethanol neurotoxicity. 1. Direct effects on replicating astrocyte. Neurobehav Toxicol Teratol 8: 11–15, 1986

    PubMed  Google Scholar 

  28. Ishioka N, Oda T, Natake Y, Kurioka S: Analysis and separation of Synaptosomal membrane proteins. Neurochem Res 15: 475–481, 1990

    PubMed  Google Scholar 

  29. Strocchi P, Brown BA, Young JD, Bonventre JA, Gilbert JM: The characterization of tubulin in CNS membrane fractions. J Neurochem 37: 1295–1307, 1981

    PubMed  Google Scholar 

  30. Kelly PT, Cotman CW: Synaptic proteins: Characterization of tubulin and actin and identification of a distinct postsynaptic density polypeptide. J Cell Biol 79: 173–183, 1978

    PubMed  Google Scholar 

  31. Isobe T, Ishioka N, Okuyama T: Isolation and characterization of des(Ala-Lys)calmodulin in porcine brain. Biochem Biophys Res Commun 102: 279–286, 1981

    PubMed  Google Scholar 

  32. Takahashi Y, Manabe T, Kadoya T, Ishioka N, Isobe T, Okyama T: Two-dimensional electrophoresis of bovine brain proteins soluble and insoluble fractions. In: H Hirai (ed.) Electrophoresis '83. Walter de Gruyter and Co., Berlin, New York, 1986, pp 263–269

    Google Scholar 

  33. Rawat AK: Effects of maternal ethanol consumption on hepatic lipid biosynthesis in foetal and neonatal rats. Biochem J 174: 213–219, 1978

    PubMed  Google Scholar 

  34. Sigh SP, Snyder AK: Ethanol ingestion during pregnancy. Effects on pregnant rats and their offspring. J Nutr 112: 98–103, 1982

    PubMed  Google Scholar 

  35. Abel EL, Dintcheff BA: Effects of prenatal alcohol exposure on growth and development in rats. Pharmacol Exp Ther 207: 906–921, 1982

    Google Scholar 

  36. Ledig M, Kopp P, Mandel P: Alcohol effect on superoxide dismutase activity in nervous tissue. Advances in Bio Sci 71: 79–86, 1988

    Google Scholar 

  37. Rappoport DA, Fritz RR, Myers JL: Nucleic acids. In: A Lajtha (ed.) Handbook of Neurochemistry, Vol. 1. Plenum Publishing Corp., New York, 1969, pp 101–119

    Google Scholar 

  38. Jarlstedt J, Hamberger A: Experimental alcoholism in rats. Effect of acute ethanol intoxication on thein vitro incorporation of [3H] leucine into neuronal and glial proteins. J Neurochem 19: 2299–2306, 1972

    PubMed  Google Scholar 

  39. Lucas M, Lamande N, Lazar M, Gros F, Legult-Demare L: Developmental expression of alpha- and gamma-enolase subunits and mRNA sequences in the mouse brain. Dev Neurosci 10: 91–98, 1988

    PubMed  Google Scholar 

  40. Berl S: Glutamine synthetase. Determination of its distribution in brain during development. Biochemistry 5: 916–922, 1966

    PubMed  Google Scholar 

  41. Hernandel AM, Bell KP, NorenBerg MD: Glutamine synthetase: Glial localization in brain. Science 195: 1356–1358, 1977

    PubMed  Google Scholar 

  42. Heron DS, Shinitzky M, Hershkowitz M, Samuel D: Lipid fluidity markedly modulates the binding of serotonin to mouse brain membranes. Proc Natl Acad Sci USA 77: 7463–7467, 1980

    PubMed  Google Scholar 

  43. Goldstein DB: Alcohol and cellular membranes. Alcohol and the developing brain. In: U Rydberg, C Alling, J Engel (eds.) Raven Press, New York, 1985, pp 19–26

    Google Scholar 

  44. Chin JH, Goldstein DB: Effects of low concentrations of ethanol on the fluidity of spin-labeled erythrocyte and brain membranes. Mol Pharmacol 13: 435–441, 1977

    PubMed  Google Scholar 

  45. Dow KE, Riopelle RJ: Specific effects of ethanol on neurite promoting proteoglycans of neuronal origin. Brain Res 508: 40–45, 1990

    PubMed  Google Scholar 

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Author notes

  1. Phanithi Prakasa Babu

    Present address: Department of Zoology, Sri Venkatesware University, 317 502, Tirupati, India

  2. L. Ratna Kumari

    Present address: Genetique Medicale, Hospital Sainte-Justine, 3175 Cote Sainte-Catherine, H3T 1C5, Montreal, Quebec, Canada

Authors and Affiliations

  1. School of Life Sciences, University of Hyderabad, 500 134, Hyderabad, India

    Phanithi Prakasa Babu, L. Ratna Kumari & Mohan C. Vemuri

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Babu, P.P., Kumari, L.R. & Vemuri, M.C. Differential changes in cell morphology, macromolecular composition and membrane protein profiles of neurons and astrocytes in chronic ethanol treated rats. Mol Cell Biochem 130, 29–40 (1994). https://doi.org/10.1007/BF01084265

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  • DOI: https://doi.org/10.1007/BF01084265

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