Abstract
Immunofluorescence staining with antibodies to tubulin, neurofilaments and glial filaments was used to study the effects of methylmercury on the differentiation of retinoic acid-induced embryonal carcinoma cells into neurons and astroglia and on the cytoskeleton of these neuroectodermal derivatives. Methylmercury did not prevent undifferentiated embryonal carcinoma cells from developing into neurons and glia. Treatment of committed embryonal carcinoma cells with methylmercury doses exceeding 1 μM resulted in the formation of neurons with abnormal morphologies. In differentiated cultures, microtubules were the first cytoskeletal element to be affected. Their disassembly was time- and concentration-dependent. Microtubules in glial cells and in neuronal perikarya were more sensitive than those in neuronal processes. Neurofilaments and glial filaments appeared relatively insensitive to methylmercury treatment but showed reorganization after complete disassembly of the microtubules. The data demonstrate 1) the sensitivity of microtubules of both neurons and glia to methylmercury-induced depolymerization, and 2) the heterogeneous response of neuronal
Similar content being viewed by others
Abbreviations
- α-MEM:
-
alpha minimal essential medium
- EC:
-
embryonal carcinoma cells
- FCS:
-
fetal calf serum
- MAP:
-
microtubule-associated protein
- MeHg:
-
methylmercury
- RA:
-
retinoic acid
References
AITCHISON, W.A. and BROWN, D.L. (1986). Duplication of the flagellar apparatus and cytoskeletal microtubule system in the alga POLYTOMELLA. Cell Mot. Cytoskeleton 6:122–127.
BLACK, M.M. and GREENE, L.A. (1982). Changes in the colchicine susceptibility of microtubules associated with neurite outgrowth: Studies with nerve growth factor-responsive PC12 pheochromocytoma cells. J. Cell Biol. 95:379–386.
BRADY, S.T. and BLACK, M.M. (1986).Axonal transport of microtubule proteins: Cytotypic variation of tubulin and MAPs in neurons. Ann. N.Y. Acad. Sci. 466:199–217.
CAVANAGH, J.B. and CHEN, F.C.K. (1971). Amino acid incorporation in protein during the “silent phase” before organo-mercury and p-bromophenylacetylurea neuropathy in rats. Acta. Neuropath. (Berlin) 19:216–244.
CHOI, B.H. (1983). Mercury and abnormal development of the fetal brain. In: NEUROBIOLOGY OF THE TRACE ELEMENTS, Vol. I, pp. 197–235. Dreosti and R. Smith, Editors. The Human Press.
CHOI, B.H., LAPHAM, L.W., AMIN-ZAKI, L. and SALEEM, T. (1978). Abnormal migration, deranged cerebral cortical organization, and diffuse white matter astrocytosis of human fetal brain: A major effect of methylmercury poisoning IN UTERO. J. Neuropathol. Exp. Neurol. 37:719–733.
CUMMINGS, R., BURGOYNE, R.D. and LYTTON, N.A. (1983). Axonal subpopulation in the central nervous system demonstrated using monoclonal antibodies against α-tubulin. Eur. J. Cell Biol. 31:241–248.
CUMMINGS, R., BURGOYNE, R.D. and LYTTON, N.A. (1984). Immunocytochemical demonstration of α-tubulin modifications during axonal maturation in the cerebellar cortex. J. Cell Biol. 98:347–351.
GOZES, I. (1981). Multiple tubulin forms are expressed by a single neurone. Nature (London) 294:477–480.
GUNDERSEN, G.G. and BULINSKI, J.C. (1986). Microtubule arrays in differentiated cells contain elevated levels of a post-translationally modified form of α-tubulin. Eur. J. Cell Biol. 42:288–294.
HARADA, M. (1978). Congenital Minamata disease: Intrauterine methylmercury poisoning. Teratology 18:285–288.
IMURA, N., MIURA, K., INODAWA, M. and NAKADA, S. (1980). Mechanism of methylmercury cytotoxicity by biochemical and morphological experiments using cultured cells. Toxicology 17:214–254.
JONES-VILLENEUVE, E.M.V., McBURNEY, M.W., ROGERS, K.A. and KALNINS, V.I. (1982). Retinoic acid induces embryonal carcinoma cells to differentiate into neurons and glial cells. J. Cell Biol. 94: 253–262.
KEATES, R.A.B. and YOTT, B. (1984). Inhibition of microtubule polymerization by micromolar concentrations of mercury (II). Can. J. Biochem. Cell Biol. 62:814–818.
MATSUMOTO, H., KOYA, G. and TAKEUCHI, T. (1965). Fetal Minamata disease. A neuropathological study of two cases of intrauterine intoxication by methylmercury compound. J. Neuropath. Exp. Neurol. 24:563–574.
McBURNEY, M.W., JONES-VILLENEUVE, E.M.V., EDWARDS, M.K.S. and ANDERSON, P.J. (1982). Control of muscle and neuronal differentiation in a cultured embryonal carcinoma cell line. Nature (London) 299: 165–167.
MIURA, K., INOKAWA, M. and IMURA, N. (1984). Effects of methylmercury and some metal ions on microtubule networks in mouse glioma cells and IN VITRO tubulin polymerization. Toxicol. Appl. Pharmacol. 73:218–231.
OLSON, F.C. and MASSARO, E.J. (1977). Effects of methylmercury on murine fetal amino acid uptake, protein synthesis and palate closure. Teratology 16:187–194.
PRASAD, K.N., NOBLES, E. and RAMANUJAM, M. (1979). Differential sensitivities of glioma cells and neuroblastoma cells to methylmercury toxicity in cultures. Environm. Res. 19:189–201.
RAFF, C., MILLER, R.H. and NOBEL, M. (1983). A glial progenitor cell that develops IN VITRO into an astrocyte or an oligodendrocyte depending on culture medium. Nature (London) 303:390–396.
REUHL, K.R. and CHANG, L.W. (1979). Effects of methylmercury on the development of the nervous system: A review. Neurotoxicology 1:21–55.
RODIER, P.M., ASCHER, M. and SAGER, P.R. (1984). Mitotic arrest in the developing CNS after prenatal exposure to methylmercury. Neurobehav. Toxicol. Teratol. 6:379–385.
RUDNICKI, M.A. and McBURNEY, M.W. (1987). Cell culture methods and induction of differentiation of embryonal carcinoma cell lines. In: PRACTICAL APPROACH BOOK ON TERATOCARCINOMAA AND EMBRYONIC STEM CELLS. Ed. E. Robertson. IRL Press, Oxford.
SAGER, P.R., DOHERTY, R.A. and OLMSTED, J.B. (1983). Interation of methylmercury with microtubules in cultures cells and in vitro. Exp. Cell Res. 146:127–137.
SAGER, P.R. and SYVERSEN, T.L.M. (1984). Differential responses to methylmercury exposure and recovery in neuroblastoma and glioma cells and fibroblasts. Exper. Neur. 85:371–382.
SAGER, P.R. and SYVERSEN, T.L.M. (1986). Disruption of microtubules by methylmercury. In: THE CYTOSKELETON: A TARGET FOR TOXIC AGENTS, pp. 97–116. T.W. Clarkson, P.R. Sager and T.L.M. Syversen (eds). Plenum Press, N.Y.
TAKEUCHI, T. (1977). Pathology of fetal Minamata disease. Pediatrician 6:69–87.
THRASHER, J.D. (1972). The effect of four mercury compounds on the generation time and cell division in TETRAHYMENA PYRIFORMIS, WH14. Environm. Res. 5:443–450.
VOGEL, D.G., MARGOLIS, R.L. and MOTTET, N.K. (1985). The effects of methylmercury binding to microtubules. Toxicol. Appl. Pharmacol. 80:473–486.
WASTENEYS, G.O., CADRIN, M., REUHL, K.R. and BROWN, D.L. (1987). The effects of methylmercury on the cytoskeleton of murine embryonal carcinoma cells. Cell Biol. Toxicol. 4.
YAMADA, K.M., SPOONER, B.S. and WESSELLS, N.K.J. (1970). Axon growth: roles of microfilaments and microtubules. Proc. Natl. Acad. Sci. U.S.A. 66:1206–1212.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cadrin, M., Wasteneys, G.O., Jones-Villeneuve, E.M.V. et al. Effects of methylmercury on retinoic acid-induced neuroectodermal derivatives of embryonal carcinoma cells. Cell Biol Toxicol 4, 61–80 (1988). https://doi.org/10.1007/BF00141287
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00141287