An enzymatically enhanced recording technique for Limulus ventral photoreceptors: physiology, biochemistry, and morphology.
Enzymatic treatments that facilitated whole-cell electrophysiological recordings were used on Limulus ventral photoreceptor cells. Ventral optic nerves were treated with either collagenase or collagenase, papain, and trypsin. Either treatment greatly increased the ease of making whole-cell recordings of transmembrane potentials. Light responses obtained from enzyme-treated photoreceptor cells were nearly identical to results obtained without enzyme treatment and compared favorably to in vivo recordings of light responses from the compound lateral eye. Enzyme-treated cells also responded to applied octopamine, as do untreated cells, with an increased phosphorylation of a 122-kD protein. This suggests that the external receptors and internal biochemical machinery required for at least one second-messenger cascade are present after enzyme treatment. The morphological integrity of enzyme-treated photoreceptor cells was examined with light microscopy as well as with scanning and transmission electron microscopy. In general, we found that each enzyme treatment greatly reduced the integrity of the layers of glial cells that surround the photoreceptor cells thereby making these cells easily accessible for whole-cell recordings of transmembrane potentials. The morphology of the rhabdomere was normal after enzymatic degradation of the adjacent glial covering.
Zhang
H J
HJ
Department of Zoology, University of Maine, Orono 04469.
Jinks
R N
RN
Wishart
A C
AC
Battelle
B A
BA
Chamberlain
S C
SC
Fahrenbach
W H
WH
Kass
L
L
D000818:Animals; D002469:Cell Separation / Q000379:methods; D017364:Collagenases; D004594:Electrophysiology; D005260:Female; D006737:Horseshoe Crabs; D008297:Male; D008564:Membrane Potentials / Q000502:physiology*; D009655:Octopamine / Q000494:pharmacology; D009799:Ocular Physiological Phenomena; D010206:Papain; D010766:Phosphorylation; D017956:Photoreceptor Cells, Invertebrate / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D015290:Second Messenger Systems / Q000502:physiology; D014357:Trypsin
D009655:Octopamine; D014357:Trypsin; D010206:Papain; D017364:Collagenases