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Photoreceptor cells dissociated from the compound lateral eye of the horseshoe crab, Limulus polyphemus, II: Function

Published online by Cambridge University Press:  02 June 2009

W. J. Brad Hanna
Affiliation:
Biophysics Interdepartmental Group, Department of Physics, University of Guelph, Guelph, Ontario, Canada
Edwin C. Johnson
Affiliation:
Department of Physiology, Marshall University School of Medicine, Huntington
Deborah Chaves
Affiliation:
Biophysics Interdepartmental Group, Department of Physics, University of Guelph, Guelph, Ontario, Canada
George H. Renninger
Affiliation:
Biophysics Interdepartmental Group, Department of Physics, University of Guelph, Guelph, Ontario, Canada

Abstract

A combination of enzymatic digestions and mechanical disruption was used to isolate photoreceptor cells from the compound lateral eye of the horseshoe crab, Limulus polyphemus. The cells were maintained in a culture medium and tested for function using whole-cell and cell-attached patch configurations of the gigaseal technique. The cells dissociated from the eye generated spontaneous voltage and current bumps in the dark, and depolarized in a graded fashion to increasing intensities of light over several decades, producing responses similar to those of cells in vivo. Currents evoked during voltage clamp were similar to those in ventral photoreceptor cells of Limulus, although transient currents in the dark- and light-activated currents were smaller in isolated lateral eye cells, perhaps because of the slow speed and spatial nonuniformity of the clamp in these large cells. In addition to isolated cells, dissociation of the compound eye produced small clusters of cells and isolated ommatidia which were also tested for function. Comparison of the electrical characteristics of isolated cells with those of cells in small clusters and in their ommatidial matrix suggests that the electrical junctions normally connecting photoreceptor cells within an ommatidium are functional in the latter groups, but not in isolated cells. Cell-attached patches of rhabdomeral membrane of isolated cells contained light-activated channels, resembling those observed in ventral photoreceptor cells, but no voltage-activated channels. Similar patches of arhabdomeral membrane contained voltage-activated channels, but no light-activated channels. We conclude that this preparation is suitable for studies of processes involved in generating the light response in invertebrate photoreceptor cells.

Type
Research Articles
Copyright
Copyright © Cambridge University Press 1993

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