Gonzalez-Bellido Paloma T, Wardill Trevor J, Kostyleva Ripsik, Meinertzhagen Ian A, Juusola Mikko
Department of Biomedical Science, University of Sheffield, Sheffield S10 2TN, UK.
J Neurosci. 2009 Nov 11;29(45):14199-210. doi: 10.1523/JNEUROSCI.2873-09.2009.
shibire(ts1), a temperature-sensitive mutation of the Drosophila gene encoding a Dynamin orthologue, blocks vesicle endocytosis and thus synaptic transmission, at elevated, or restrictive temperatures. By targeted Gal4 expression, UAS-shibire(ts1) has been used to dissect neuronal circuits. We investigated the effects of UAS-shibire(ts1) overexpression in Drosophila photoreceptors at permissive (19 degrees C) and restrictive (31 degrees C) temperatures. At 19 degrees C, overexpression of UAS-shi(ts1) causes decelerated phototransduction and reduced neurotransmitter release. This phenotype is exacerbated with dark adaptation, age and in white mutants. Photoreceptors overexpressing UAS-shibire(ts1) contain terminals with widespread vacuolated mitochondria, reduced numbers of vesicles and bundled microtubules. Immuno-electron microscopy reveals that the latter are dynamin coated. Further, the microtubule phenotype is not restricted to photoreceptors, as UAS-shibire(ts1) overexpression in lamina cells also bundles microtubules. We conclude that dynamin has multiple functions that are interrupted by UAS-shibire(ts1) overexpression in Drosophila photoreceptors, destabilizing their neural communication irreversibly at previously reported permissive temperatures.
失活(ts1)是果蝇中一个编码发动蛋白同源物的基因的温度敏感突变体,在升高的或限制的温度下会阻断囊泡内吞作用,从而阻断突触传递。通过靶向Gal4表达,UAS-失活(ts1)已被用于剖析神经回路。我们研究了在允许温度(19摄氏度)和限制温度(31摄氏度)下,UAS-失活(ts1)在果蝇光感受器中过表达的影响。在19摄氏度时,UAS-失活(ts1)的过表达会导致光转导减速和神经递质释放减少。这种表型在暗适应、衰老和白色突变体中会加剧。过表达UAS-失活(ts1)的光感受器的终末含有广泛空泡化的线粒体、数量减少的囊泡和成束的微管。免疫电子显微镜显示,后者被发动蛋白包被。此外,微管表型并不局限于光感受器,因为在层细胞中过表达UAS-失活(ts1)也会使微管成束。我们得出结论,发动蛋白具有多种功能,这些功能在果蝇光感受器中会被UAS-失活(ts1)的过表达所中断,在先前报道的允许温度下不可逆地破坏其神经通讯。