The preparation and use of dispersed cells from fat body of Locusta migratoria in a filtration plate assay for adipokinetic peptides.
A method is described for the preparation of metabolically active and hormone-sensitive dispersed cells from fat body of Locusta migratoria and their subsequent use in a filtration plate assay. This assay measures the uptake of radiolabelled acetate into the cells, and the use of dispersed cells reduces the time and labor involved in preparing dose-response curves to different test peptides and increases the precision of the estimate of potency. As a consequence of the reduced variability, fewer replicates per dose are required and, when operated as a plate assay, up to eight dose-response curves can be constructed per plate. The method described here for measuring metabolite uptake into cells and thus determining the potencies of adipokinetic hormones could be applied to the assay of both catabolic (glucagon-like) and anabolic (insulin-like) substances, not only in other insects, but also in mammalian cells. It could be used, therefore, as a suitable screening method for novel active compounds.
Lee
M J
MJ
Department of Biology, Birkbeck College, University of London, England.
Goldsworthy
G J
GJ
D000085:Acetates / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002918:Chymotrypsin; D005216:Fat Body / Q000166:cytology* / Q000378:metabolism; D006110:Grasshoppers / Q000166:cytology*; D007700:Kinetics; D010455:Peptides / Q000032:analysis*
D000085:Acetates; D010455:Peptides; D002918:Chymotrypsin