Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1972 Sep;10(3):537–544. doi: 10.1128/jvi.10.3.537-544.1972

Inhibition of Sindbis Virus Release by Media of Low Ionic Strength: an Electron Microscope Study

Marilynn R F Waite 1,2, Dennis T Brown 1,2, Elmer R Pfefferkorn 1,2
PMCID: PMC356495  PMID: 4672394

Abstract

Release of Sindbis virus from infected cells is inhibited by lowering the ionic strength of the medium. To determine the nature of the inhibited step, we examined, by electron microscopy, both freeze-etched and thin-sectioned preparations which had been fixed with either glutaraldehyde or formaldehyde. Inhibitory medium had two different effects on Sindbis virus release: virus budding was partially inhibited, and those virions which did mature were precipitated on the surface of the cell. Freeze-etched, inhibited cells showed very few viral buds. After shift to normal medium, the number of budding virions increased dramatically, far exceeding the quantity found in normal controls. Thus, low ionic strength medium clearly inhibited an early stage of virus maturation. The results were the same regardless of the fixative. Thin sections of glutaraldehyde-fixed, inhibited cells contained large extracellular aggregates of mature virus which were not present in similar, formaldehyde-fixed preparations. Fixation of radioactively-labeled, inhibited cultures revealed that approximately half of the virus that could be released from inhibited cells by raising the ionic strength of the medium could also be released by formaldehyde, but not by glutaraldehyde. This fraction probably represents mature virus attached to the cell surface by the ionic conditions.

Full text

PDF
539

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Acheson N. H., Tamm I. Replication of Semliki Forest virus: an electron microscopic study. Virology. 1967 May;32(1):128–143. doi: 10.1016/0042-6822(67)90261-9. [DOI] [PubMed] [Google Scholar]
  2. Brinkley B. R., Murphy P., Richardson L. C. Procedure for embedding in situ selected cells cultured in vitro. J Cell Biol. 1967 Oct;35(1):279–283. doi: 10.1083/jcb.35.1.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown D. T., Waite M. R., Pfefferkorn E. R. Morphology and morphogenesis of Sindbis virus as seen with freeze-etching techniques. J Virol. 1972 Sep;10(3):524–536. doi: 10.1128/jvi.10.3.524-536.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark A. W., Branton D. Fracture faces in frozen outer segments from the guinea pig retina. Z Zellforsch Mikrosk Anat. 1968;91(4):586–603. doi: 10.1007/BF00455276. [DOI] [PubMed] [Google Scholar]
  5. Hopwood D. A comparison of the crosslinking abilities of glutaraldehyde, formaldehyde and alpha-hydroxyadipaldehyde with bovine serum albumin and casein. Histochemie. 1969;17(2):151–161. doi: 10.1007/BF00277781. [DOI] [PubMed] [Google Scholar]
  6. PFEFFERKORN E. R., CLIFFORD R. L. THE ORIGIN OF THE PROTEIN OF SINDBIS VIRUS. Virology. 1964 Jun;23:217–223. doi: 10.1016/0042-6822(64)90285-5. [DOI] [PubMed] [Google Scholar]
  7. PFEFFERKORN E. R., HUNTER H. S. THE SOURCE OF THE RIBONUCLEIC ACID AND PHOSPHOLIPID OF SINDBIS VIRUS. Virology. 1963 Jul;20:446–456. doi: 10.1016/0042-6822(63)90093-x. [DOI] [PubMed] [Google Scholar]
  8. STEERE R. L. Electron microscopy of structural detail in frozen biological specimens. J Biophys Biochem Cytol. 1957 Jan 25;3(1):45–60. doi: 10.1083/jcb.3.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Scheele C. M., Pfefferkorn E. R. Virus-specific proteins synthesized in cells infected with RNA+ temperature-sensitive mutants of Sindbis virus. J Virol. 1970 Mar;5(3):329–337. doi: 10.1128/jvi.5.3.329-337.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Waite M. R., Pfefferkorn E. R. Effect of altered osmotic pressure on the growth of Sindbis virus. J Virol. 1968 Jul;2(7):759–760. doi: 10.1128/jvi.2.7.759-760.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Waite M. R., Pfefferkorn E. R. Inhibition of Sindbis virus production by media of low ionic strength: intracellular events and requirements for reversal. J Virol. 1970 Jan;5(1):60–71. doi: 10.1128/jvi.5.1.60-71.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES