Cell Biology and Toxicology

, Volume 11, Issue 1, pp 29–36 | Cite as

Methylmercury-injury effect on tube formation by cultured human vascular endothelial cells

  • T. Kishimoto
  • T. Oguri
  • M. Tada


The effect of methylmercury chloride (MeHg) on growth and tube formation by cultured human umbilical vein endothelial cells (HUVECs) was investigated. HUVECs were collected by enzymatic digestion with collagenase. Precultivation of HUVECs with MeHg at concentrations of 1.0–50.0 μmol/L exerted negligible effects on the viable cell number, while the viable cell number was slightly reduced at 100 μmol/L and fell to zero at concentrations exceeding 500.0 μmol/L MeHg. The viable cell number was depressed in a concentration-dependent manner. Tube formation was studied by culturing the cells on gelled basement membrane matrix (Matrigel). Treatment of HUVECs with 0.1–5.0 μmol/L MeHg for 24 h inhibited tube formation dose-dependently. Fetal bovine serum (FBS) increased tube formation in a dose-dependent manner, with half-maximum stimulation of tube formation at approximately 3.4% FBS. The length of tube formation decreased time-dependently at concentrations of 0.1 and 1.0 μmol/L MeHg. Pretreatment of Matrigel with 1 μmol/L MeHg before the cell seeding reduced the tube formation by HUVECs. These results suggest that the growth and tube formation by HUVECs is susceptible to MeHg cytotoxicity, and that MeHg could be injurious to endothelial cell function.

Key words

endothelial cell methylmercury tube formation basement membrane matrix 



methylmercury chloride


human umbilical vein endothelial cells


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Folkman J. Toward an understanding of angiogenesis: search and discovery. Perspect Biol Med. 1985;29:10–36.Google Scholar
  2. Geelen JAG, Dormans JAMA, Verhoef A. The early effects of methylmercury on the developing rat brain. Acta Neuropathol. 1990;80:432–8.Google Scholar
  3. Grant DS, Tashiro K, Segui-Real B, Yamada Y, Martin GR, Kieinman HK. Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures in vitro. Cell. 1989;58:933–43.Google Scholar
  4. Hunter D, Russell DS. Focal cerebral and cerebellar atrophy in a human subject due to organic mercury compounds. J Neurol Neurosurg Psychiatry. 1954;17:235–41.Google Scholar
  5. Hunter D, Bomford RR, Russell DS. Poisoning by methylmercury compounds. Q J Med. 1940;9:193–213.Google Scholar
  6. Jackson JC, Jenkins LK. Type I collagen fibrils promote rapid vascular tube formation upon contact with the apical side of cultured endothelium. Exp Cell Res. 1991;192:319–23.Google Scholar
  7. Kasama H, Itoh K, Omata S, Sugano H. Differential effects of methylmercury on the synthesis of protein species in dorsal root ganglia of the rat. Arch Toxicol. 1989;63:226–30.Google Scholar
  8. Kinsella JL, Grant DS, Weeks BS, Kleinman HK. Protein kinase C regulates endothelial cell tube formation on basement membrane matrix, Matrigel. Exp Cell Res. 1992;199:56–62.Google Scholar
  9. Kishimoto T. The effect of antifebriles on human vascular endothelial cells at high temperature in cell culture. Jpn J Biometeor. 1987;24:117–24. (In Japanese with English abstract).Google Scholar
  10. Kishimoto T, Fukuzawa Y, Tada M. Studies on human endothelial cells; basic attempt at primary cell culture. Bull Shimane Med Univ. 1982;5:23–8. (In Japanese with English abstract).Google Scholar
  11. Kubota Y, Kleinman HK, Martin GR, Lawley TJ. Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol. 1988;107:1589–98.Google Scholar
  12. Luscher TF, Vanhoutte FM. Endothelium-dependent contractions to acetylcholine in the aorta of the spontaneously hypertensive rat. Hypertension. 1986;8:344–8.Google Scholar
  13. Luscher TF, Raji L, Vanhoutte PM. Endothelium-dependent vascular responses in normotensive and hypertensive Dahl rats. Hypertension. 1987;9:157–63.Google Scholar
  14. Lushcer TF, Diederich D, Weber E. Endothelium-dependent responses in carotid and renal arteries of normotensive and hypertensive rats. Hypertension, 1988;11:573–8.Google Scholar
  15. Madri JA, Williams SK, Wyatt T, Mezzio C. Capillary endothelial cell culture: phenotypic modulation by matrix components. J Cell Biol. 1983;97:153–65.Google Scholar
  16. Madri JA, Pratt BM. Angiogenesis: In: Clark RF, Henson P, eds. The molecular and cellular biology of wound healing. New York: Plenum; 1987:377–58.Google Scholar
  17. Madri JA, Pratt BM, Tucker AM. Phenotypic modulation of endothelial cells by transforming growth factor-β depends upon the composition and organization of the extracellular matrix. J Cell Biol. 1988;106:1375–84.Google Scholar
  18. Montesano R. Cell-extracellular matrix interactions in morphogenesis: an in vitro approach. Experientia. 1986;42:977–85.Google Scholar
  19. Montesano R, Orci L, Vassali P. In vitro rapid organization of endothelial cells into capillary-like networks is promoted by collagen matrices. J Cell Biol 1983;97:1648–52.Google Scholar
  20. O'Kusky J. Methylmercury poisoning of the developing nervous system: morphological changes in neuronal mitochondria. Acta Neuropathol (Berl.) 1983;61:116–22.Google Scholar
  21. Omata S, Sugano H. Biochemical studies on methylmercury poisoning of nervous tissues. In: Tsubaki T, Takahashi H, eds. Recent advances in Minamata disease studies. Tokyo: Kodansha; 1986:150–69.Google Scholar
  22. Oyake Y. Pathology of organic mercury intoxication occurring in the basin Agano. Shinkei Kenkyu no Shinpo. 1969;13:108–12. (In Japanese with English abstract).Google Scholar
  23. Oyake Y, Tanaka M, Kubo H, Chichibu M. Neurological studies on organic mercury intoxication, with special reference to distribution of mercury granules. Shinkei Kenkyu no Shinpo. 1966;10:744–50. (In Japanese with English abstract).Google Scholar
  24. Oyanagi K, Ohama E, Ikuta F. The auditory system in methyl mercurial intoxication: a neuropathological investigation on 14 autopsy cases in Niigata, Japan. Acta Neuropathol. 1989;77:561–8.Google Scholar
  25. Ross R, Glomset AJ. The pathogenesis of atherosclerosis. New Engl J Med. 1976;295:369–420, 420–5.Google Scholar
  26. Shiraki H. Neuropathological aspects of organic mercury intoxication, including Minamata disease. Handb Clin Neurol. 1979;36:83–145.Google Scholar
  27. Sugimoto T, Tobian L, Ganguli MC. High potassium diets protect against dysfunction of endothelial cells in strokeprone spontaneously hypertensive rats. Hypertension. 1988;11:579–85.Google Scholar
  28. Takeuchi T. Neuropathology of Minamata disease in Kumamoto: especially at the chronic stage. In: Roizin I, Shiraki H, Grcevic N, eds. Neurotoxicology. New York: Raven Press; 1977:235–46.Google Scholar
  29. Takeuchi T, Morikawa N, Matsumoto H, Shiraishi Y. A pathological study of Minamata disease in Japan. Acta Neuropathol. 1962;2:40–57.Google Scholar
  30. Tsubaki T, Hirota K, Shirakawa K, Kondo K, Sato T. Clinical, epidemiological, and toxicological studies on methylmercury poisoning. In: Plaa GL, Cuncan WIAM, eds. Proceedings of the first international congress on toxicology. New York: Academic Press; 1978:339–57.Google Scholar
  31. Yoshino Y, Nakano K. Biochemical changes in the brain in rats poisoned with an alkylmercury compound, with special reference to the inhibition of protein synthesis in brain cortex slices. J Neurochem. 1966;13:1223–30.Google Scholar
  32. Zucker RM, Elstein KH, Easterling RE, Massaro EJ. Flow cytometric analysis of the mechanism of methylmercury cytotoxicity. Am J Pathol. 1990;137:187–198.Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • T. Kishimoto
    • 1
  • T. Oguri
    • 1
  • M. Tada
    • 1
  1. 1.Department of Environmental MedicineShimane Medical UniversityIzumoJapan

Personalised recommendations