Summary
Semliki Forest virus infected Aedes albopictus cells were used to investigate virus induced cell-cell fusion. It was shown by a novel method that cell-cell fusion was completed within approximately 5 minutes after triggering the fusion event by low pH. This method consists of fixing fusing cells with glutaraldehyde and microinjecting the highly fluroescent and rapidly diffusing dye Lucifer yellow. In contrast, polykaryon formation, the usually used criterion to measure cell-cell fusion occurred only within 30 minutes. Furthermore, it was shown that potassium cyanide, a potent inhibitor of polykaryon formation in the described system, inhibits an early step of membrane-membrane fusion of neighbouring cells.
Keywords: Potassium, Cyanide, Glutaraldehyde, Infected Cell, Potent Inhibitor
References
- 1.Finkeldey W. Ueber Riesenzellbefunde in den Gaumenmandeln, zugleich ein Beitrag zur Histopathologie der Mandelveränderungen im Maserninkubationsstadium. Virchows Arch Pathol Anat Physiol Klin Med. 1931;281:323–329. [Google Scholar]
- 2.Henle G, Deinhardt F, Girardi A. Cytolytic effects of mumps virus in tissue cultures of epithelial cells. Proc Soc Exp Biol Med. 1954;87:386–393. doi: 10.3181/00379727-87-21390. [DOI] [PubMed] [Google Scholar]
- 3.Igarashi A. Isolation of a Singh's Aedes albopictus cell clone sensitive to Dengue and Chikungunya viruses. J Gen Virol. 1978;40:531–544. doi: 10.1099/0022-1317-40-3-531. [DOI] [PubMed] [Google Scholar]
- 4.Kempf C, Michael MR, Kohler U, Koblet H. Semliki Forest virus-induced polykaryon formation is an ATP-dependent event. Arch Virol. 1987;95:111–122. doi: 10.1007/BF01311338. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Koblet H, Kempf C, Kohler U, Omar A. Conformational changes at pH 6 on the cell surface of Semliki Forest virus-infected Aedes albopictus cells. Virology. 1985;143:334–336. doi: 10.1016/0042-6822(85)90120-5. [DOI] [PubMed] [Google Scholar]
- 6.Koblet H, Omar A, Kempf C. Fusion of alphavirus infected mosquito cells. In: Yunker CE, editor. Arbovirus cultivation in arthropod cells in vitro. Boca Raton, Florida, in press: CRC Press; 1987. [Google Scholar]
- 7.Okada Y. Analysis of giant polynuclear cell formation caused by HVJ virus from Ehrlich's ascites tumor cells. I. Microscopic observation of giant polynuclear cell formation. Exp Cell Res. 1962;26:98–107. doi: 10.1016/0014-4827(62)90205-7. [DOI] [PubMed] [Google Scholar]
- 8.Okada Y. Analysis of giant polynuclear cell formation caused by HVJ virus from Ehrlich's ascites tumor cells. III. Relationship between cell condition and fusion reaction or cell degeneration reaction. Exp Cell Res. 1962;26:119–128. doi: 10.1016/0014-4827(62)90207-0. [DOI] [PubMed] [Google Scholar]
- 9.Oshiro LS, Schieble JH, Lenette EH. Electron microscopic studies of a coronavirus. J Gen Virol. 1971;12:161–168. doi: 10.1099/0022-1317-12-2-161. [DOI] [PubMed] [Google Scholar]
- 10.Poste G, Nicholson GL. Membrane fusion, cell surface reviews, vol 5. Amsterdam: Elsevier/North Holland; 1978. [Google Scholar]
- 11.Roizman B. Polykaryocytosis. Cold Spring Harbor Symp Quant Biol. 1962;27:327–342. doi: 10.1101/sqb.1962.027.001.031. [DOI] [PubMed] [Google Scholar]
- 12.Stewart WW. Lucifer dyes-highly fluorescent dyes for biological tracing. Nature. 1981;292:17–21. doi: 10.1038/292017a0. [DOI] [PubMed] [Google Scholar]
- 13.White J, Matlin K, Helenius A. Cell fusion by Semliki Forest, influenza, and vesicular stomatitis virus. J Cell Biol. 1981;89:674–679. doi: 10.1083/jcb.89.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]