Jacobs Journal of Anesthesiology and Research

Genetic Analysis of the Resistant and Sensitive Mutants of Drosophila Melanogaster to Diethylether Anesthesia

*Yoshiharu Tanaka
Department Of Earth And Life Sciences, Osaka Prefecture University, Osaka, Japan

*Corresponding Author:
Yoshiharu Tanaka
Department Of Earth And Life Sciences, Osaka Prefecture University, Osaka, Japan
Email:yoshitan@las.osakafu-u.ac.jp

Published on: 2018-06-01

Abstract

Abstract We are investigating which kinds of genes are related to diethylether anesthesia using many resistant and sensitive mutants of Drosophila melanogaster to diethylether. A resistant mutant strain EthAR223 and a sensitive mutant strain ethas319 were made artificially by inserting a P-element, one of transposon, into the chromosome. The strains were out-crossed by white strains that have normal sensitivity to diethylether, to replace most of the EthAR223 or ethas319 chromosome by the chromosome of the white strain, except the P-element insertion region. Both outcross lines showed the properties of EthAR223 or ethas319 rather than the white strains, suggesting the altered sensitivities of the EthAR223 and ethas319 were caused by the gene that were inserted by the P-element, not by mutation of other genes. To isolate and identify the gene inserted by the P-element, the genome DNA of the ethas319 was prepared and the 3 kb sized DNA region was amplified by the polymerase chain reaction (PCR) method. The amplified DNA was sequenced and the sequence was found to have a homology with a part of a genomic clone AC019820 that was one of DNA sequence data provided by DNA Data Bank of Japan (DDBJ). The sequence has a gene that expands approximately 30 kb, has at least 14 exon sequences, and supposed to encode at least 413 amino acid sequence. This gene was supposed as the Drosophila counter part of the casein kinase 1, gamma 3 subunit (CK1- γ3) gene because it had a high homology with the rat CK1- γ3 by the sequence comparison. With the further analysis of the DNA sequence, the altered sensitivity of the ethas319 was caused by the P-element insertion at the non-coded region of the CK1- γ 3 gene that probably related to the nervous functions.

Keywords

Diethylether; Anesthesia; Fly; Mutant; Gene; Casein Kinase; Nervous System

Introduction

The P-element, one of the transposable gene (transposon) is able to insert into chromosomes of Drosophila melanogaster at random, sometimes causing mutations on a variety of genes. By screening the objective mutant and by the transposon tagging method that is to clone the gene related to the phenotype easily using the P-element as a scaffold, the genes related to diethylether anesthesia would be found to help revealing of anesthesia mechanisms. By this method we obtained and analyzed 19 P-element inserted strains having altered sensitivities to diethylether anesthesia, suggesting many genes would relate to the sensitivity to diethylether anesthesia. Among them, the para locus that encodes a voltage-dependent sodium channel gene and the crc locus that encodes a calcium storage protein calreticulin gene are closely involved in the hypersensitivity to diethylether anesthesia. In this study, we analyze other two mutant strains EthAR223 and ethas319 to understand the mechanisms of anesthesia in detail by clarifying more genes related to sensitivity to anesthesia.