Agonist stimulation provokes dendritic and axonal dopamine D(1) receptor redistribution in primary cultures of striatal neurons.
To investigate the influence of neurotransmitter on G-protein-coupled receptor trafficking and compartimentalization in neurons, we have developed a model of primary neuronal cultures from fetal rat striatum on which we have studied the cellular and subcellular distribution and trafficking of the D(1) dopaminergic receptor. This receptor is known to be somatodendritic and axonal targeted in vivo, mostly to extrasynaptic locations. Immunohistochemical studies at the light and electron microscopic levels showed that, in cultures, the D(1) dopaminergic receptor is expressed in the absence of dopamine stimulation. The pattern of D(1) dopaminergic receptor immunostaining after stimulation by the D(1) dopaminergic receptor agonist SKF 82958 (1 microM) is dramatically modified with a decrease of the number of labeled D(1) dopaminergic receptor puncta (-40%) and an increase of their size in both dendrites (+120%) and axons (+240%). Seven hours after removal of the agonist, return to normal pattern was observed. The D(1) dopaminergic receptor antagonist SCH 23390 (2 microM) abolishes the effect of SKF 82958. Electron microscopy demonstrated, in dendrites, a translocation of the labeling from the plasma membrane to endosomes. Axonal D(1) dopaminergic receptor redistribution after acute stimulation indicates that the D(1) dopaminergic receptor is membrane targeted and responsive to stimulation. These results validate primary culture of striatal neurons to study subcellular localization and intraneuronal trafficking of G-protein-coupled receptors. This preparation will be useful to address various questions concerning the behavior and the trafficking of these receptors in neurons in relation to the neurotransmitter environment.
Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5541, Laboratoire d'Histologie-Embryologie, Universite V. Segalen-Bordeaux II, 146 rue Leo Saignat, 33076 Cedex, Bordeaux, France. marie-laure.martin-negrier@umr5541.u-bordeaux2.fr
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M
Martin-Negrier
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G
Charron
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B
Bloch
D000818:Animals; D001369:Axons / Q000187:drug effects / Q000378:metabolism*; D001552:Benzazepines / Q000494:pharmacology; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000166:cytology; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism*; D018491:Dopamine Agonists / Q000494:pharmacology; D004622:Embryo, Mammalian; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017447:Receptors, Dopamine D1 / Q000187:drug effects / Q000378:metabolism*
D001552:Benzazepines; D018491:Dopamine Agonists; D012333:RNA, Messenger; D017447:Receptors, Dopamine D1; C071262:SK&F 82958