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Characterization of Mitotic Neurons Derived From Adult Rat Hypothalamus and Brain Stem
- Source :
- Journal of Neurophysiology. 87:1076-1085
- Publication Year :
- 2002
- Publisher :
- American Physiological Society, 2002.
-
Abstract
- Embryonic or neonatal rat neurons retain plasticity and are readily grown in tissue culture, but neurons of the adult brain were thought to be terminally differentiated and therefore difficult to culture. Recent studies, however, suggest that it may be possible to culture differentiated neurons from the hippocampus of adult rats. We modified these procedures to grow differentiated neurons from adult rat hypothalamus and brain stem. At day 7 in tissue culture and beyond, the predominant cell types in hypothalamic and brain stem cultures had a stellate morphology and could be subdivided into two distinct groups, one of which stained with antibodies to the immature neuron marker α-internexin, while the other stained with the astrocyte marker GFAP. The α-internexin positive cells were mitotic and grew to form a characteristic two-dimensional cellular network. These α-internexin positive cells coimmunostained for the neuronal markers MAP2, type III β-tubulin, and tau, and also bound tetanus toxin, but were negative for the oligodendrocyte marker GalC and also for the neurofilament triplet proteins NF-L, NF-M, and NF-H, markers of more mature neurons. Patch-clamp analysis of these α-internexin positive cells revealed small Ca2+ currents with a peak current of −0.5 ± 0.1 pA/pF at a membrane potential of −20 mV ( n = 5) and half-maximal activation at −30 mV ( n = 5). Na+ currents with a peak current density of −154.5 ± 49.8 pA/pF at a membrane potential of −15 mV ( n = 5) were also present. We also show that these cells can be frozen and regrown in tissue culture and that they can be efficiently infected by viral vectors. These cells therefore have the immunological and electrophysiological properties of immature mitotic neurons and should be useful in a variety of future studies of neuronal differentiation and function.
- Subjects :
- Male
Patch-Clamp Techniques
Physiology
Hypothalamus
Mitosis
tau Proteins
Biology
Rats, Inbred WKY
Membrane Potentials
Rats, Sprague-Dawley
Tissue culture
Intermediate Filament Proteins
Neurofilament Proteins
Tubulin
Glial Fibrillary Acidic Protein
Animals
Cells, Cultured
Neurons
Neonatal rat
Stem Cells
General Neuroscience
Age Factors
Antibodies, Monoclonal
Cell Differentiation
Embryonic stem cell
Rats
Female
Carrier Proteins
Microtubule-Associated Proteins
Neuroscience
Brain Stem
Subjects
Details
- ISSN :
- 15221598 and 00223077
- Volume :
- 87
- Database :
- OpenAIRE
- Journal :
- Journal of Neurophysiology
- Accession number :
- edsair.doi.dedup.....f9ac575b38b2a7a4823eed786d437ce6