The Journal of Neuroscience, July 15, 1998, 18(14):5253-5263
Membrane Current Induced by Protein Kinase C Activators in
Rhabdomeric Photoreceptors: Implications for Visual Excitation
Maria del Pilar
Gomez and
Enrico
Nasi
Department of Physiology, Boston University School of Medicine,
Boston, Massachusetts 02118, and Marine Biological Laboratory, Woods
Hole, Massachusetts 02543
Visual excitation in rhabdomeric photoreceptors is thought to be
mediated by activation of a light-regulated phospholipase C (PLC) and
the consequent hydrolysis of phosphatidylinositol bisphosphate. Whereas
much attention has been devoted to inositol trisphosphate
(IP3) production and intracellular
Ca2+ release, little is known about the possible
role of the DAG branch in the generation of the light response. We have
tested the effect of chemically distinct surrogates of DAG on isolated
Lima photoreceptors. Application of the phorbol ester
PMA (0.5-10 µM) or the alkaloid (
)-indolactam (20-100
µM) from a holding potential of
50 mV elicited an
inward current, several hundred picoamperes in amplitude, accompanied
by a pronounced increase in membrane conductance. The stereoisomers
4
-PMA and (+)-indolactam were both inactive, arguing for the
specificity of the effects. Elevation of cytosolic Ca2+ by intracellular dialysis accelerated this
current, whereas chelerythrine antagonized it, suggesting the
involvement of PKC. The reversal potential of the membrane current
induced by PKC activators was approximately +10 mV; replacement of
extracellular Na with impermeant N-methyl-D-glucamine decreased its amplitude and
shifted the reversal potential in the negative direction. Stimulation
by PMA and (
)-indolactam was accompanied by a pronounced depression
of light responsiveness; conversely, steady illumination reduced the
size of the current elicited by these PKC activators. Taken together,
these results support the notion that the DAG branch of the PLC
cascade, in addition to its suggested participation in visual
adaptation, may play a role in the activation of the photoresponse or a
component thereof, probably in synergy with IP3-mediated
Ca2+ release.
Key words:
visual excitation; PKC; diacylglycerol; rhabdomeric
photoreceptors; light-dependent conductance; calcium
Copyright © 1998 Society for Neuroscience 0270-6474/98/18145253-11$05.00/0