Positive and negative feedback regulation of choline acetyltransferase mRNA levels in Drosophila: a study using temperature-sensitive mutants and embryo cell cultures

Abstract

Steady state levels of the mRNA coding for the neurotransmitter biosynthetic enzyme, acetylCoA-choline- O-acetyltransferase (ChAT, EC 2.3.1.6), were measured in wild type Drosophila and two temperature-sensitive mutants (Cha ts1 and Cha ts2) using the RNase protection method. At a permissive temperature the relative amounts of ChAT mRNA were: wild type:Cha ts1:Cha ts2 = 1:2.09 (± 0.39):3.37 (± 0.57) (mean ± S.E.M.) indicating that mutant flies may compensate, for making a thermolabile form of enzyme, by producing and/or maintaining higher levels of ChAT mRNA. At a restrictive temperature the ChAT mRNA levels decreased in both mutants and increased in wild type flies. The regulatory mechanism(s) responsible for increasing ChAT mRNA in wild type flies appears to have failed in the mutants at high temperature. Steady state mRNA levels were also measured in embryonic cell cultures prepared from wild type embryos. Cultures grown in the presence of two pharmacologic agents (carbamylcholine and d-tubocurarine) which should interfere with cholinergic neurotransmission, showed less mRNA resulting from a decrease in levels of ChAT gene transcription. Our results imply that neurotransmission and the rate of neurotransmitter biosynthetic enzyme gene transcription are coupled for the cholinergic system in Drosophila.