Dissociation of photoreceptors from whole heads of the fruit fly, Drosophila melanogaster.
Photoreceptor cells that were mostly free of extracellular material and suitable for most electrophysiological study procedures were dissociated from whole heads of the fruit fly, Drosophila melanogaster, by a simple "smash" technique employing gentle chopping by a razor blade through Parafilm sheets. A variety of commonly available proteolytic and glycolytic digestion enzymes were tested as additions to the basic dissociation procedure described. With the aid of Nomarski interference contrast optics, periodic acid-Schiff staining, and fluorescent labeling and microscopy methods, it was determined that proteolytic enzymatic digestion does little to enhance the dissociation procedure, and instead, often damages the cells that one is attempting to recover. Unexpectedly, certain glycolytic enzymes, when added to the basic procedure, appear to enhance the recovery of intact viable Drosophila photoreceptors that are stripped of most extracellular material. Based on these results, a hypothesis concerning the biochemical nature of the extracellular matrix of the Drosophila retina is proposed. Drosophila photoreceptors are an interesting model system for the study of invertebrate phototransduction and photoreceptor cell biology because of their many well-characterized mutant strains. The technique described here should produce clean viable photoreceptors or ommatidia that respond to light, and that are suitable for patch clamping or cell culture.
Ziemba
S E
SE
Department of Biological Sciences, Wayne State University, Detroit, MI 48202, USA.
Saks
S
S
Janviriya
Y
Y
Stephenson
R S
RS
D000818:Animals; D003584:Cytological Techniques; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000737:chemistry*; D004798:Enzymes / Q000494:pharmacology; D005109:Extracellular Matrix / Q000737:chemistry; D006257:Head; D010503:Periodic Acid-Schiff Reaction; D017956:Photoreceptor Cells, Invertebrate / Q000737:chemistry / Q000502:physiology; D014020:Tissue Extracts
D004798:Enzymes; D014020:Tissue Extracts