Isolation of monoaminergic synaptosomes from rat brain by immunomagnetophoresis.
Monoaminergic synaptosomes have been isolated and purified from rat brain by immunomagnetophoresis. This novel technique uses magnetic beads to which Protein A is bound. Noradrenergic, dopaminergic, and serotonergic synaptosomes (previously cell-surface labelled with anti-dopamine-beta-hydroxylase, anti-tyrosine hydroxylase, and anti-tryptophan hydroxylase, respectively) may be isolated in a highly purified state. The synaptosomal subpopulations are recovered in a viable metabolic state and show glucose-stimulated respiration and Ca2(+)-dependent neurotransmitter release. A novel subtype of dopamine-beta-hydroxylase was found in dopaminergic terminals. No evidence for glutamate corelease from monoaminergic synaptosomes was obtained.
Docherty
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Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, England.
Bradford
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Cash
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Ehret
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Maitre
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Joh
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D000818:Animals; D001679:Biogenic Amines / Q000378:metabolism*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure*; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007158:Immunologic Techniques; D008280:Magnetics / Q000379:methods; D018377:Neurotransmitter Agents / Q000378:metabolism; D010101:Oxygen Consumption; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D013574:Synaptosomes / Q000378:metabolism / Q000648:ultrastructure*; D014365:Tryptophan Hydroxylase / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D001679:Biogenic Amines; D005904:Glial Fibrillary Acidic Protein; D018377:Neurotransmitter Agents; D014446:Tyrosine 3-Monooxygenase; D014365:Tryptophan Hydroxylase; D004299:Dopamine beta-Hydroxylase; D011188:Potassium