Tanaka Y, Gamo S
Department of Life Sciences, College of Integrated Arts and Sciences, Osaka Prefecture University, 1-1 Gakuen-Cho, Sakai, 599-8531, Osaka, Japan
Colloids Surf B Biointerfaces. 2001 Sep;22(1):39-53. doi: 10.1016/s0927-7765(01)00156-4.
The para locus of Drosophila melanogaster encodes the alpha subunit of a voltage-sensitive sodium channel. Many of the mutants develop paralysis at the high temperature (37 degrees C) and are hypersensitive to diethylether anesthesia. We examined whether the two aspects of the phenotype are mediated by a same mechanism that involves the sodium channel molecule by investigating properties of the three para alleles (para(hd838), para(ts1) and para(ts3)). Larvae of the all para strains showed almost normal sensitivities to diethylether anesthesia while adult flies of them showed hypersensitivities to that in the following manner: para(hd838)<==para(ts1)<para(ts3)<Canton-S. Larvae of the two para strains showed hypersensitivities to heat-induced paralysis in the following manner: para(ts1)=para(ts3)<para(hd838)=Canton-S, while adult flies of the all para strains showed hypersensitivities to that in the following manner: para(ts1)=para(ts3)<para(hd838)<Canton-S. The distinct phenotype of para(hd838) from para(ts1) or para(ts3) observed in the larval and adult heat-induced paralysis, would be the reflection of the difference of the mutation sites between in para(hd838) and para(ts1) or para(ts3). In addition, because the rank of sensitivity in the adult anesthesia and heat-induced paralysis were reversed between para(hd838) and para(ts1) or para(ts3), the mechanisms of diethylether anesthesia and heat-induced paralysis are not the same. There would be the anesthesia-specific mechanisms because the sensitivity to anesthesia became more remarkable in para(hd838) than in para(ts1) or para(ts3). para(hd838) has an insertion of P-element, one of a transposable element, in the second intron of the para sodium channel gene. Excision of the P-element from the para locus of the para(hd838) conferred the flies the wild-typic phenotype and reduction of the para gene in para(hd838) dramatically enhanced the hypersensitivity to anesthesia, suggesting that the phenotype of the strain was caused exclusively by the mutation of the para locus. The susceptibility to anesthesia also depends on the temperature at which the flies were assayed, not at which they were cultured, but the dependence on the assay temperature was smaller than that on the genotype. By these findings we could assure that the sensitivity to diethylether anesthesia of para(hd838) primarily depends on the genotypes of the sodium channel gene rather than other reasons such as the fluidity of the membranes.
果蝇的para基因座编码一种电压敏感钠通道的α亚基。许多该基因的突变体在高温(37摄氏度)下会发生麻痹,并且对乙醚麻醉高度敏感。我们通过研究三个para等位基因(para(hd838)、para(ts1)和para(ts3))的特性,来探究该表型的这两个方面是否由涉及钠通道分子的相同机制介导。所有para品系的幼虫对乙醚麻醉的敏感性几乎正常,而它们的成虫对乙醚麻醉表现出如下的高度敏感性:para(hd838)<==para(ts1)<para(ts3)<Canton-S。两个para品系的幼虫对热诱导麻痹表现出如下的高度敏感性:para(ts1)=para(ts3)<para(hd838)=Canton-S,而所有para品系的成虫对热诱导麻痹表现出如下的高度敏感性:para(ts1)=para(ts3)<para(hd838)<Canton-S。在幼虫和成虫热诱导麻痹中观察到的para(hd838)与para(ts1)或para(ts3)不同的表型,可能反映了para(hd838)与para(ts1)或para(ts3)之间突变位点的差异。此外,由于para(hd838)与para(ts1)或para(ts3)在成虫麻醉和热诱导麻痹中的敏感性排序相反,所以乙醚麻醉和热诱导麻痹的机制并不相同。可能存在麻醉特异性机制,因为para(hd838)对麻醉的敏感性比para(ts1)或para(ts3)更显著。para(hd838)在para钠通道基因的第二个内含子中有一个P因子插入,P因子是一种转座元件。从para(hd838)的para基因座切除P因子使果蝇具有野生型表型,并且在para(hd838)中降低para基因的表达会显著增强对麻醉的高度敏感性,这表明该品系的表型完全是由para基因座的突变引起的。对麻醉的易感性还取决于检测果蝇时的温度,而不是它们培养时的温度,但对检测温度的依赖性小于对基因型的依赖性。通过这些发现我们可以确定,para(hd838)对乙醚麻醉的敏感性主要取决于钠通道基因的基因型,而非其他原因,如细胞膜的流动性。