Primary culture of striatal neurons: a model of choice for pharmacological and biochemical studies of neurotransmitter receptors.
Striatal neurons were cultured from fetal mouse brain and maintained in serum-free medium for 14-21 days in vitro (DIV). A double coating of culture dishes with polyornithine and fetal calf serum was needed in order to obtain synaptic differentiation. Synaptic vesicles were present in axon terminals as well as in varicosities along extended axons. The presence of differentiated synapses was confirmed by the immunostaining of the preparation with synapsin I antibody. After 13 days in vitro synapsin I was present in axonal varicosities and particularly concentrated at contact points between axonal terminals and postsynaptic sites on adjacent axons or perikarya. On a surface of 429 mm2 on which 2211 cells were observed under phase contrast microscopy only 7% were stained with an antibody against GFAP (glial fibrillary acidic protein). One or two days after the formation of differentiated synapses (11 DIV), a Ca2+-dependent liberation of GABA was observed. These cultures are an excellent model for studying the coupling of some neurotransmitter receptors with an adenylate cyclase. In particular using this preparation we were able to demonstrate that dopamine (D2) and serotonin-(5-HT1) receptors are negatively coupled with an adenylate cyclase. These cultures are also an excellent model to study the coupling of some neurotransmitter receptors with inositol phosphate producing enzymes. We demonstrated for the first time that the quisqualate subtype of glutamate receptors is able to increase inositol phosphate production in striatal neurons.
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000166:cytology / Q000502:physiology*; D005333:Fetus; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D007295:Inositol Phosphates / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D009474:Neurons / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D018377:Neurotransmitter Agents / Q000494:pharmacology; D017981:Receptors, Neurotransmitter / Q000502:physiology*; D013569:Synapses / Q000166:cytology / Q000502:physiology
D005904:Glial Fibrillary Acidic Protein; D007295:Inositol Phosphates; D018377:Neurotransmitter Agents; D017981:Receptors, Neurotransmitter