P19 cells differentiate into glutamatergic and glutamate-responsive neurons in vitro

The neurotransmitter l-glutamate has been associated with a number of developmental events within the central nervous system including synaptogenesis and the refinement of topographically ordered neural maps. As a model for studying such events at the molecular level, we have examined the expression...

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Bibliographic Details
Published in:Neuroscience 1997-09, Vol.80 (2), p.487-499
Main Authors: MacPherson, P.A, Jones, S, Pawson, P.A, Marshall, K.C, McBurney, M.W
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Blotting, Northern
Cell Differentiation
Cell Line
Electrophysiology
embryonal carcinoma
Fluorescent Antibody Technique, Direct
Fundamental and applied biological sciences. Psychology
gamma-Aminobutyric Acid - physiology
Glutamic Acid - physiology
Immunohistochemistry
Isolated neuron and nerve. Neuroglia
Neurons - physiology
Neurons - ultrastructure
neurotransmitter
Patch-Clamp Techniques
Rats
receptor
Receptors, Glutamate - metabolism
Receptors, Glutamate - physiology
retinoic acid
Synapses - physiology
Synapses - ultrastructure
Vertebrates: nervous system and sense organs
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Summary:The neurotransmitter l-glutamate has been associated with a number of developmental events within the central nervous system including synaptogenesis and the refinement of topographically ordered neural maps. As a model for studying such events at the molecular level, we have examined the expression of glutamate and glutamate receptors in neurons that develop from P19 cells in response to retinoids. We report here that many P19-derived neurons do contain glutamate in secretory vesicles and that this glutamate appears to function as a neurotransmitter. The neurotransmitter GABA is also present in these cultures and both glutamate and GABA appeared to co-localize in some neuronal processes. Both neurotransmitters were released from the neurons in response to membrane depolarization. These neurons also express various glutamate receptor subunits including GluR1, GluR4 and NMDAR1 as detected by immunological methods. Using whole-cell patch-clamping, we have recorded spontaneous postsynaptic potentials which increase in both amplitude and frequency with time in culture and which are sensitive to the glutamate antagonist kynurenic acid. Thus, P19-derived neurons mature in culture and form electrically active neural networks involving glutamate and glutamate receptors.
ISSN:0306-4522
1873-7544
DOI:10.1016/S0306-4522(97)00102-4