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Cell Biology and Toxicology

, Volume 9, Issue 4, pp 359–375 | Cite as

Characterization of cytoskeletal and neuronal markers in micromass cultures of rat embryonic midbrain cells

  • S. G. Whittaker
  • J. T. Wroble
  • S. M. Silbernagel
  • E. M. Faustman
Article

Abstract

Micromass cultures of rat embryonic midbrain cells were characterized with regard to the immunolocalization of neuronal and cytoskeletal markers. Cells taken from gestational day-12 embryos and cultured for 5 days in vitro comprise at least two morphologically distinct cell types:-fibroblast-like cells and neurons. Antibodies to the following markers yieldedpreferential staining of neuronal cells: A2B5 (GQ ganglioside), ψ-aminobutyric acid (GABA), microtubule-associated protein 2 (MAP2), MIPS, neuron-specific enolase (NSE), neural cell adhesion molecule (N-CAM), and tau. Antibodies to β-tubulin, χ-neu, MAPI, and neurofilament (NF-H) stained both neuronal and fibroblast-like cells. Antibodies to glial fibrillary acidicprotein (GFAP) and vimentin failed to immunoreact with any cells in day-5 CNS cultures. SDS-PAGE and Western analysis were employed to determine the specificity of the antibodies and determine the electrophoretic profiles of the markers. We conclude that thepattern of neuronal differentiation in CNS micromass cultures exhibits certain similarities to that observed in vivo. In addition, certain markers identified in this study may be ofpotential utility as (1) biomarkers of chemically-induced developmental neurotoxicity, and (2) indicators of diferential toxicity toward the diverse cell types that comprise the mammalian central nervous system.

Keywords

Neuronal Differentiation Neuronal Marker Neural Cell Adhesion Molecule Distinct Cell Western Analysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • S. G. Whittaker
    • 1
    • 2
  • J. T. Wroble
    • 1
  • S. M. Silbernagel
    • 1
  • E. M. Faustman
    • 1
    • 3
  1. 1.Department of Environmental HealthUniversity of WashingtonSeattleUSA
  2. 2.Department of PathologyUniversity of WashingtonSeattleUSA
  3. 3.Child Development and Mental Retardation CenterSeattleUSA

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