Culture of postnatal mesencephalic dopamine neurons on an astrocyte monolayer.
This unit presents a protocol for primary culture of postnatal mesencephalic dopamine neurons grown on an astrocyte monolayer, which can be used to investigate cellular and molecular mechanisms regulating dopamine neuron function. Using this in vitro approach, dopamine neurons survive for an extended period of time and establish functional axon terminals and dendrites that display properties similar to those observed in vivo and in brain slices. An alternate protocol is provided for a microculture system in which astrocytes are grown on a spatially limited surface where single or small groups of dopamine neurons develop. Under such conditions, isolated neurons establish synaptic contacts, or autapses, onto their own somatodendritic compartment, thus facilitating morphological and physiological experiments.
Fasano
Caroline
C
Département de Pharmacologie, Faculté de Médecine, Université de Montréal, Québec, Canada.
Thibault
Dominic
D
Trudeau
Louis-Eric
LE
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D001369:Axons / Q000378:metabolism / Q000648:ultrastructure; D002450:Cell Communication / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003712:Dendrites / Q000378:metabolism / Q000648:ultrastructure; D004298:Dopamine / Q000378:metabolism*; D051379:Mice; D009434:Neural Pathways / Q000166:cytology / Q000254:growth & development; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D013378:Substantia Nigra / Q000166:cytology* / Q000254:growth & development; D013569:Synapses / Q000502:physiology / Q000648:ultrastructure; D017557:Ventral Tegmental Area / Q000166:cytology* / Q000254:growth & development
D004298:Dopamine