Microdialysis as a tool for in vivo study of dopamine transporter function in rat brains.
The role of dopamine as a major modulator of CNS function is well-known, and the homeostasis of dopamine is considered to be of major importance in the pathogenesis of several psychiatric and neurological diseases. Few methods are currently available for in vivo study of dopamine transporter function, which regulates extracellular levels of dopamine. Adapting the 'indicator diffusion' method applied to the microdialysis technique, we present here a suitable method for this functional investigation. We measured the cellular extraction of [3H]-MPP+, which is known to accumulate in the dopaminergic neurones through the DAT in the rat striatum, using [14C]-mannitol as reference substance characterized by absence of cellular accumulation. The cellular extraction was 0.41 and was almost abolished in the presence of the dopamine-uptake inhibitor cocaine, reaching 0.07. This suggested that extraction of [3H]-MPP+ was due to cellular uptake by dopamine transporters. Tissue analysis confirmed that [3H]-MPP+ was internalized in cells and that such transport was stopped by cocaine. Moreover, [3H]-MPP+ extraction was dramatically decreased after lesioning the nigro-striatal pathway with 6-hydroxydopamine, whereas [14C]-mannitol extraction was unchanged. It is concluded that the presented method can be used to study the functioning of the dopamine transporter in live animals.
Zimmer
L
L
CERMEP Biomedical Cyclotron, Lyon, France.
Kodas
E
E
Guilloteau
D
D
Garreau
L
L
Besnard
J
J
Chalon
S
S
D015655:1-Methyl-4-phenylpyridinium / Q000494:pharmacology; D000818:Animals; D002250:Carbon Radioisotopes; D002352:Carrier Proteins / Q000187:drug effects / Q000378:metabolism*; D003042:Cocaine / Q000494:pharmacology; D003714:Denervation / Q000009:adverse effects; D050483:Dopamine Plasma Membrane Transport Proteins; D008297:Male; D008353:Mannitol / Q000494:pharmacology; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D017551:Microdialysis / Q000379:methods*; D017072:Neostriatum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009419:Nerve Tissue Proteins; D016627:Oxidopamine; D011869:Radioligand Assay / Q000706:statistics & numerical data; D051381:Rats; D017208:Rats, Wistar; D012588:Scintillation Counting / Q000706:statistics & numerical data; D014316:Tritium
D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; C493216:Slc6a3 protein, rat; D014316:Tritium; D008353:Mannitol; D016627:Oxidopamine; D003042:Cocaine; D015655:1-Methyl-4-phenylpyridinium