Single Drosophila ommatidium dissection and imaging

TY - JOUR

T1 - Single Drosophila ommatidium dissection and imaging

AU - Volpi, Vera

AU - Mackay, D

AU - Fanto, Manolis

PY - 2011/10/26

Y1 - 2011/10/26

N2 - The fruit fly Drosophila melanogaster has made invaluable contributions to neuroscience research and has been used widely as a model for neurodegenerative diseases because of its powerful genetics(1). The fly eye in particular has been the organ of choice for neurodegeneration research, being the most accessible and life-dispensable part of the Drosophila nervous system. However the major caveat of intact eyes is the difficulty, because of the intense autofluorescence of the pigment, in imaging intracellular events, such as autophagy dynamics(2), which are paramount to understanding of neurodegeneration. We have recently used the dissection and culture of single ommatidia(3) that has been essential for our understanding of autophagic dysfunctions in a fly model of Dentatorubro-Pallidoluysian Atrophy (DRPLA)(3, 4). We now report a comprehensive description of this technique (Fig. 1), adapted from electrophysiological studies(5), which is likely to expand dramatically the possibility of fly models for neurodegeneration. This method can be adapted to image live subcellular events and to monitor effective drug administration onto photoreceptor cells (Fig. 2). If used in combination with mosaic techniques(6-8), the responses of genetically different cells can be assayed in parallel (Fig. 2).

AB - The fruit fly Drosophila melanogaster has made invaluable contributions to neuroscience research and has been used widely as a model for neurodegenerative diseases because of its powerful genetics(1). The fly eye in particular has been the organ of choice for neurodegeneration research, being the most accessible and life-dispensable part of the Drosophila nervous system. However the major caveat of intact eyes is the difficulty, because of the intense autofluorescence of the pigment, in imaging intracellular events, such as autophagy dynamics(2), which are paramount to understanding of neurodegeneration. We have recently used the dissection and culture of single ommatidia(3) that has been essential for our understanding of autophagic dysfunctions in a fly model of Dentatorubro-Pallidoluysian Atrophy (DRPLA)(3, 4). We now report a comprehensive description of this technique (Fig. 1), adapted from electrophysiological studies(5), which is likely to expand dramatically the possibility of fly models for neurodegeneration. This method can be adapted to image live subcellular events and to monitor effective drug administration onto photoreceptor cells (Fig. 2). If used in combination with mosaic techniques(6-8), the responses of genetically different cells can be assayed in parallel (Fig. 2).

U2 - 10.3791/2882

DO - 10.3791/2882

M3 - Article

SN - 1940-087X

VL - 54

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

M1 - e2882

ER -