TY - JOUR
T1 - Synapse formation and establishment of neuronal polarity by P19 embryonic carcinoma cells and embryonic stem cells
AU - Finley, Michael F.A.
AU - Kulkarni, Nita
AU - Huettner, James E.
PY - 1996/2/1
Y1 - 1996/2/1
N2 - A number of different cell lines that exhibit a partial neuronal phenotype have been identified, but in many cases the full extent of their neuronal differentiation has not been directly addressed by functional studies. We have used electrophysiology and immunofluorescence to examine the formation of synapses and the development of neuronal polarity by murine embryonic stem (ES) cells and the mouse P19 embryonic carcinoma cell line. Within 2-3 weeks after induction by retinoic acid, subsets of P19 and ES cells formed excitatory synapses, mediated by glutamate receptors, or inhibitory synapses, mediated by receptors for GABA or glycine. In ES-cell cultures, both NMDA and non-NMDA receptors contributed to the excitatory postsynaptic response. Staining with antibodies to growth-associated protein-43 and microtubule- associated protein-2 revealed segregation of immunoreactivity into separate axonal and somato-dendritic compartments, respectively. Consistent with our physiological evidence for synapse formation, intense punctate staining was observed with antibodies to the synaptic vesicle proteins synapsin, SV2, and synaptophysin. These results demonstrate the in vitro acquisition by pluripotent cell lines of neuronal polarity and functional synaptic transmission that is characteristic of CNS neurons.
AB - A number of different cell lines that exhibit a partial neuronal phenotype have been identified, but in many cases the full extent of their neuronal differentiation has not been directly addressed by functional studies. We have used electrophysiology and immunofluorescence to examine the formation of synapses and the development of neuronal polarity by murine embryonic stem (ES) cells and the mouse P19 embryonic carcinoma cell line. Within 2-3 weeks after induction by retinoic acid, subsets of P19 and ES cells formed excitatory synapses, mediated by glutamate receptors, or inhibitory synapses, mediated by receptors for GABA or glycine. In ES-cell cultures, both NMDA and non-NMDA receptors contributed to the excitatory postsynaptic response. Staining with antibodies to growth-associated protein-43 and microtubule- associated protein-2 revealed segregation of immunoreactivity into separate axonal and somato-dendritic compartments, respectively. Consistent with our physiological evidence for synapse formation, intense punctate staining was observed with antibodies to the synaptic vesicle proteins synapsin, SV2, and synaptophysin. These results demonstrate the in vitro acquisition by pluripotent cell lines of neuronal polarity and functional synaptic transmission that is characteristic of CNS neurons.
KW - NMDA receptors
KW - embryonic carcinoma cells
KW - embryonic stem cells
KW - neuronal differentiation
KW - neuronal polarity
KW - synaptic transmission
UR - http://www.scopus.com/inward/record.url?scp=0029985691&partnerID=8YFLogxK
U2 - 10.1523/jneurosci.16-03-01056.1996
DO - 10.1523/jneurosci.16-03-01056.1996
M3 - Article
C2 - 8558234
AN - SCOPUS:0029985691
VL - 16
SP - 1056
EP - 1065
JO - Journal of Neuroscience
JF - Journal of Neuroscience
SN - 0270-6474
IS - 3
ER -