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Mouse peripherin isoforms.

F Landon1, A Wolff, B de Néchaud

  • 1Université Paris 6, faculté de médecine Pitié-Salpêtrière, CNRS UMR 7000, France. landon@idf.ext.jussieu.fr

Biology of the Cell
|January 2, 2001
PubMed
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Mouse peripherin gene alternative splicing produces multiple mRNA transcripts and protein isoforms. These peripherin isoforms, including minor ones, are present in nervous tissues and contribute to intermediate filament networks.

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Cell Biology

Background:

  • The mouse peripherin gene undergoes alternative splicing to produce distinct mRNAs.
  • These mRNAs translate into three protein isoforms: Pe-58 (major), Pe-56 (shorter C-terminus), and Pe-61 (inserted central domain).

Purpose of the Study:

  • To investigate the simultaneous occurrence of multiple peripherin transcripts in murine nervous tissues and neuroblastoma cell lines.
  • To characterize the peripherin isoforms and their cellular localization within intermediate filament networks.

Main Methods:

  • Polymerase Chain Reaction (PCR) amplification to detect alternative splicing events.
  • Recombinant expression of peripherin isoforms in E. coli.
  • Generation and immunoadsorption of specific rabbit antisera against peripherin sequences.

Related Experiment Videos

  • Western blot analysis and two-dimensional electrophoresis for isoform localization and charge variant identification.

    • Main Results:

    • Multiple peripherin transcripts were confirmed in murine nervous tissues and neuroblastoma cell lines.
    • Peripherin isoforms were localized in neuroblastoma cell lysates and detergent-insoluble fractions.
    • Each isoform exhibited several charge variants.
    • Antibodies against peripherin isoforms decorated the intermediate filament network in NB2a cells.

    Conclusions:

    • Minor peripherin isoforms, previously less characterized, are present and functional in neuronal cells.
    • These isoforms actively participate in the formation and maintenance of the intermediate filament network.