Measurement of tryptophan hydroxylase activity in rat pineal glands and pinealocytes using an HPLC assay with electrochemical detection.
A method for measuring tryptophan hydroxylase activity by assaying the product 5-HTP using high-performance liquid chromatography (HPLC) with electrochemical detection is described. A nocturnal elevation (80%) in rat pineal gland tryptophan hydroxylase activity was detected. Experiments on isolated rat pinealocytes with the protein synthesis inhibitor cycloheximide indicate that tryptophan hydroxylase turns over rapidly in these cells. This method will be valuable in studies of the adrenergic mechanisms regulating pineal tryptophan hydroxylase activity.
Sugden
D
D
National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.
Grady
R
R
Mefford
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IN
D006916:5-Hydroxytryptophan / Q000032:analysis*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004563:Electrochemistry; D005260:Female; D010870:Pineal Gland / Q000166:cytology / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D014365:Tryptophan Hydroxylase / Q000378:metabolism*
D006916:5-Hydroxytryptophan; D014365:Tryptophan Hydroxylase