Molecular and immunocytochemical characterization of primary neuronal cultures from adult rat brain: Differential expression of neuronal and glial protein markers

Balmiki Ray, Jason A. Bailey, Sumit Sarkar, Debomoy K. Lahiri

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Neurobiological studies using primary neuronal cultures commonly employ fetal-derived neurons, but much less often adult brain-derived neurons. Our goal is to perform morphological and molecular characterization of primary neuronal cultures from adult rat brain, including the relative expression of neuronal and glial cell markers at different time points. We tested the hypothesis that long-term neuronal viability is compatible with glial proliferation in adult neuron culture. We examined neuron culture from adult rat brain, which was maintained at steady state up to 24 days, and characterized them on the basis of cellular, molecular and biochemical properties at different time points of the culture. We identified neuronal and glial cells by both immunocytochemical and western immunoblotting techniques using NSE and Tau as neuronal markers and GFAP as glial protein marker, which revealed the presence of predominantly neuronal cells in the initial phase of the culture and a rise in glial cells from day 12 onwards. Notably, neuronal cells were preserved in the culture along with the glial cells even at day 24. Transfection of the cultured cells with a GFP expression vector and plasmids containing a luciferase reporter gene under the control of two different gene promoters demonstrated DNA transfectability. Taken together, these results suggest a differential expression of neuronal and glial cells at different time points and long-term neuronal viability in the presence of glial proliferation. Such adult neurons serve as a suitable system for the application of neurodegeneration models and for drug target discovery in various brain disorders including Alzheimer's disease.

Original languageEnglish (US)
Pages (from-to)294-302
Number of pages9
JournalJournal of Neuroscience Methods
Volume184
Issue number2
DOIs
StatePublished - Nov 15 2009

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Neuroglia
Brain
Neurons
Proteins
Brain Diseases
Drug Discovery
Luciferases
Reporter Genes
Transfection
Cultured Cells
Alzheimer Disease
Plasmids
Western Blotting
DNA
Genes

Keywords

  • Adult neuron culture
  • Amyloid precursor protein
  • Glia
  • Neuron
  • Transfection

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Molecular and immunocytochemical characterization of primary neuronal cultures from adult rat brain : Differential expression of neuronal and glial protein markers. / Ray, Balmiki; Bailey, Jason A.; Sarkar, Sumit; Lahiri, Debomoy K.

In: Journal of Neuroscience Methods, Vol. 184, No. 2, 15.11.2009, p. 294-302.

Research output: Contribution to journalArticle

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