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Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle
11:57

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

Published on: March 22, 2013

Influenza virus infection in multinucleated skeletal myofibers

Mika Nevalainen1, Marja Nissinen, Mika Kaakinen

  • 1Institute of Biomedicine, Department of Anatomy and Cell Biology, PO Box 5000, FIN-90014 University of Oulu, Finland.

Experimental Cell Research
|April 6, 2010
PubMed

Insights

Influenza virus infection in rat muscle fibers is abortive. Viral RNA replication is confined to single nuclei, preventing spread to other nuclei within the same cell.

Area of Science:

  • Virology
  • Cell Biology
  • Skeletal Muscle Physiology

Background:

  • Influenza virus typically infects respiratory epithelial cells.
  • The behavior of influenza virus in multinucleated cells like myofibers is not well understood.

Purpose of the Study:

  • To investigate the progression of WSN influenza virus infection in isolated rat skeletal myofibers.
  • To determine the mechanisms of viral entry, replication, and spread within multinucleated muscle cells.

Main Methods:

  • Isolation of rat skeletal myofibers.
  • Infection with WSN influenza virus.
  • Analysis of viral entry kinetics, budding, and glycoprotein maturation.
  • Plaque assays for infectivity.
  • In situ hybridization for viral RNA localization and replication.
  • Dual in situ hybridization for co-localization of different viral RNAs.

Main Results:

  • WSN influenza virus entry into myofibers was delayed compared to mononucleated cells.
  • Viral budding occurred, but hemagglutinin maturation and cleavage were inefficient, leading to non-infective viral particles.
  • Viral RNA replication was restricted to one or a few nuclei within a myofiber.
  • Different viral genome segments replicated within the same nucleus.
  • Viral ribonucleoprotein particles (vRNPs) did not spread to uninfected nuclei.

Conclusions:

  • Influenza virus infection in rat skeletal myofibers is abortive due to restricted replication and lack of spread between nuclei.
  • Viral genome segments may enter and replicate as a package within a single nucleus.
  • The infection is limited to the initially infected nucleus, with proteins spreading locally but not leading to new infectious virions or nuclear infection.

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