Se. Ziemba et al., DISSOCIATION OF PHOTORECEPTORS FROM WHOLE HEADS OF THE FRUIT-FLY, DROSOPHILA-MELANOGASTER, Cell and tissue research, 280(2), 1995, pp. 473-477
Photoreceptor cells that were mostly free of extracellular material an
d suitable for most electrophysiological study procedures were dissoci
ated 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 a
nd glycolytic digestion enzymes were tested as additions to the basic
dissociation procedure described. With the aid of Nomarski interferenc
e contrast optics, periodic acid-Schiff staining, and fluorescent labe
ling and microscopy methods, it was determined that proteolytic enzyma
tic digestion does little to enhance the dissociation procedure, and i
nstead, often damages the cells that one is attempting to recover. Une
xpectedly, certain glycolytic enzymes, when added to the basic procedu
re, appear to enhance the recovery of intact viable Drosophila photore
ceptors that are stripped of most extracellular material. Based on the
se results, a hypothesis concerning the biochemical nature of the extr
acellular matrix of the Drosophila retina is proposed. Drosophila phot
oreceptors are an interesting model system for the study of invertebra
te phototransduction and photoreceptor cell biology because of their m
any well-characterized mutant strains. The technique described here sh
ould produce clean viable photoreceptors or ommatidia that respond to
light, and that are suitable for patch clamping or cell culture.