Abstract
The thermoprotective mechanisms of insects remain largely unknown. We reported the Bombyx mori contractile (cot) behavioral mutant with thermo-sensitive seizures phenotype. At elevated temperatures, the cot mutant exhibit seizures associated with strong contractions, rolling, vomiting, and a temporary lack of movement. We narrowed a region containing cot to ~268 kb by positional cloning and identified the mutant gene as Bmsei which encoded a potassium channel protein. Bmsei was present in both the cell membrane and cytoplasm in wild-type ganglia but faint in cot. Furthermore, Bmsei was markedly decreased upon high temperature treatment in cot mutant. With the RNAi method and injecting potassium channel blockers, the wild type silkworm was induced the cot phenotype. These results demonstrated that Bmsei was responsible for the cot mutant phenotype and played an important role in thermoprotection in silkworm. Meanwhile, comparative proteomic approach was used to investigate the proteomic differences. The results showed that the protein of Hsp-1 and Tn1 were significantly decreased and increased on protein level in cot mutant after thermo-stimulus, respectively. Our data provide insights into the mechanism of thermoprotection in insect. As cot phenotype closely resembles human epilepsy, cot might be a potential model for the mechanism of epilepsy in future.
Highlights
The ability to sense and respond to the ambient temperature is crucial for the survival and fitness of all animals
Using combinations of immunofluorescence, immunoblotting, RNA interference (RNAi) and injection of a potassium channel blocker, these results demonstrate that B. mori seizure gene (Bmsei) was responsible for the cot mutant phenotype
The WT (+ /+ ) did not show the epilepsy phenotype, even at 48 °C. These results suggest that cot/cot is incompletely dominant because the cot/+ heterozygote showed the cot/cot phenotype at 45 °C
Summary
The ability to sense and respond to the ambient temperature is crucial for the survival and fitness of all animals. The genes identified are responsible for larval body color (sch)[18], the larval stripe (cts)[19], blood color (Y and rb)[20,21], cocoon color (Gb)[22], pupal and moth color[23,24] (so and mln), pathology mutants (nsd-1 and nsd-2)[25,26], and moultinism (mod and nm-g)[27,28] These mutants are valuable resources for studying the genetics and gene functions of the silkworm Bombyx mori. Cot is an excellent mutant with which to study relationship between behaviors and the molecular mechanism that protect insects from the effects of high temperatures. Our results suggest that the cot mutant is a good model in which to study the behavior and the molecular mechanism of thermoprotection in insects
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