Expression of rat neurofilament proteins NF-L and NF-M in transfected non-neuronal cells

Eur J Cell Biol. 1989 Dec;50(2):475-90.

Abstract

Two cDNA clones fully encoding the rat neurofilament proteins NF-L and NF-M were subcloned into eukaryotic expression vectors behind the strong constitutive viral promoters from SV40 and Rous sarcoma viruses. Transient transfection of L tk- and Cos cell lines with these expression constructs resulted in cells expressing the neurofilament proteins in an intermediate filament-type pattern. Additionally, a putative juxtanuclear organizing center or region was observed in the transfected cells, most noticeable shortly after the transfection procedure. Stable transfections were performed on mouse L tk- and Swiss 3T6 cells using NF-L and NF-M constructs bearing an SV40 early promoter driven neomycin selectable marker. Although G418-resistant clones were recovered with both the NF-L and the NF-M constructs, only clones expressing immunofluorescently stainable amounts of NF-M were detected and established. Immunoelectron microscopic analysis revealed NF-M and vimentin proteins to be colocalized on the same intermediate filaments.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Cell Line
  • Cloning, Molecular
  • Colchicine / pharmacology
  • Cytoskeleton / analysis*
  • DNA / genetics
  • Fluorescent Antibody Technique
  • Intermediate Filament Proteins / analysis
  • Intermediate Filament Proteins / biosynthesis*
  • Intermediate Filament Proteins / genetics
  • Intermediate Filaments / analysis*
  • Intermediate Filaments / ultrastructure
  • Microscopy, Electron
  • Molecular Sequence Data
  • Neurofilament Proteins
  • Plasmids
  • Rats
  • Time Factors
  • Transfection*
  • Vimentin / analysis

Substances

  • Intermediate Filament Proteins
  • Neurofilament Proteins
  • Vimentin
  • neurofilament protein L
  • DNA
  • Colchicine