Development of translationally active mRNA for larval muscle acetylcholinesterase during ascidian embryogenesis

Relative quantities of translationally active acetylcholinesterase (acetylcholine acetylhydrolase, EC 3.1.1.7) mRNA present at various developmental stages were compared in embryos of the ascidian Ciona intestinalis. Purified RNA was tested for its translational capacity by microinjection into Xenopus laevis oocytes; the acetylcholinesterase produced was immunoprecipitated with antibody to Ciona acetylcholinesterase and enzyme activity was assayed radiometrically. With this protocol, enzyme synthesis was found to be directly related to the amount of RNA injected and to the oocyte incubation time. A functional template for acetylcholinesterase was first detected at 6 hr of development (late gastrula) and is probably present as early as 5 hr. The level of this template activity increased until the middle tail formation stage (11-12 hr after fertilization) and then remained constant until 16 hr of development (the final stage examined), 2 hr before hatching. These findings, and the results of previous actinomycin D inhibition experiments, indicate that mRNA for ascidian larval muscle acetylcholinesterase is first synthesized during gastrulation.