Catechol-O-methyltransferase activity in rat brain primary neuronal and glial cell cultures and its inhibitation by novel drugs
✍ Scribed by Ilkka Reenilä; Päivi Tuomainen; Seppo Soinila; Raimo K. Tuominen; Pekka T. Männistö
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 924 KB
- Volume
- 25
- Category
- Article
- ISSN
- 0893-6609
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✦ Synopsis
To analyze the intracellular catechol-O-methyltransferase (COMT) activity in cultured cells, a substrate (dihydroxybenzoic acid) was added to the tissue culture plates and the product (vanillic acid) was analyzed with reversed-phase HPLC with coulometric detection. The cultures, prepared from various regions of the brain, were immunohistochemically characterized. Basal COMT activities were similar in all glial cultures while in some neuron-enriched cultures the enqme activity was lower. Preincubation with nitrocatechol COMT inhibitors, entacapone or tolcapone (30-300 I&%), decreased COMT activity in all culture types with slightly different potencies. An atypical inhibitor of 0-methylation, CGP 28104, had no effect on COMT activity in any of these cultures. In conclusion: 1) this method enables straight-forward analysis of a large number of COMT activity samples in cultured cells, 2) COMT activity is almost equally distributed throughout the brain, and 3) tolcapone was a more potent COMT inhibitor than entacapone in cultures containing glial cells.