Abstract
Retinas from the scallop Pecten irradians were enzymatically dispersed, yielding a large number of isolated photoreceptors suitable for tight- seal recording. Whole-cell voltage clamp measurements demonstrated that the phototransducing machinery remained intact: quantum bumps could be elicited by dim illumination, while brighter flashes produced larger, smooth photocurrents. Single-channel currents specifically activated by light were recorded in cell-attached patches, and were almost exclusively confined to the rhabdomeric region. Their density is sufficiently high to account for the macroscopic photoresponse. Channel activation is graded with stimulus intensity in a range comparable to that of the whole-cell response, and can be recorded with illumination sufficiently dim to evoke only quantum bumps. Light-dependent channel openings are very brief, on average 1 ms or less at 20-22 degrees C, apparently not because of blockage by extracellular divalent cations. The mean open time does not change substantially with stimulus intensity. In particular, since dwell times are in the millisecond range even with the dimmest lights, the channel closing rate does not appear to be the rate-limiting step for the decay kinetics of discrete waves. The latency of the first opening after light onset is inversely related to light intensity, and the envelope of channel activity resembles the time course of the whole-cell photocurrent. Unitary currents are inward at resting potential, and have a reversal voltage similar to that of the macroscopic light response. Voltage modulates the activity of light-sensitive channels by increasing the opening rate and also by lengthening the mean open times as the patch is depolarized. The unitary conductance of the predominant class of events is approximately 48 pS, but at least one additional category of smaller- amplitude openings was observed. The relative incidence of large and small events does not appear to be related in a simple way to the state of adaptation of the cell.
Full Text
The Full Text of this article is available as a PDF (2.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Andreoli T. E., Monahan M. The interaction of polyene antibiotics with thin lipid membranes. J Gen Physiol. 1968 Aug;52(2):300–325. doi: 10.1085/jgp.52.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bacigalupo J., Chinn K., Lisman J. E. Ion channels activated by light in Limulus ventral photoreceptors. J Gen Physiol. 1986 Jan;87(1):73–89. doi: 10.1085/jgp.87.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bacigalupo J., Lisman J. E. Single-channel currents activated by light in Limulus ventral photoreceptors. Nature. 1983 Jul 21;304(5923):268–270. doi: 10.1038/304268a0. [DOI] [PubMed] [Google Scholar]
- Bader C. R., Macleish P. R., Schwartz E. A. A voltage-clamp study of the light response in solitary rods of the tiger salamander. J Physiol. 1979 Nov;296:1–26. doi: 10.1113/jphysiol.1979.sp012988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barber V. C., Evans E. M., Land M. F. The fine structure of the eye of the mollusc Pecten maximus. Z Zellforsch Mikrosk Anat. 1967;76(3):25–312. [PubMed] [Google Scholar]
- Bodoia R. D., Detwiler P. B. Patch-clamp recordings of the light-sensitive dark noise in retinal rods from the lizard and frog. J Physiol. 1985 Oct;367:183–216. doi: 10.1113/jphysiol.1985.sp015820. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Colquhoun D., Hawkes A. G. On the stochastic properties of single ion channels. Proc R Soc Lond B Biol Sci. 1981 Mar 6;211(1183):205–235. doi: 10.1098/rspb.1981.0003. [DOI] [PubMed] [Google Scholar]
- Dirnberger G., Keiper W., Schnakenberg J., Stieve H. Comparison of time constants of single channel patches, quantum bumps, and noise analysis in Limulus ventral photoreceptors. J Membr Biol. 1985;83(1-2):39–43. doi: 10.1007/BF01868736. [DOI] [PubMed] [Google Scholar]
- Dodge F. A., Jr, Knight B. W., Toyoda J. Voltage noise in Limulus visual cells. Science. 1968 Apr 5;160(3823):88–90. doi: 10.1126/science.160.3823.88. [DOI] [PubMed] [Google Scholar]
- FUORTES M. G., YEANDLE S. PROBABILITY OF OCCURRENCE OF DISCRETE POTENTIAL WAVES IN THE EYE OF LIMULUS. J Gen Physiol. 1964 Jan;47:443–463. doi: 10.1085/jgp.47.3.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fein A., Charlton J. S. Local membrane current in Limulus photoreceptors. Nature. 1975 Nov 20;258(5532):250–252. doi: 10.1038/258250a0. [DOI] [PubMed] [Google Scholar]
- Frank T. M., Fein A. The role of the inositol phosphate cascade in visual excitation of invertebrate microvillar photoreceptors. J Gen Physiol. 1991 Apr;97(4):697–723. doi: 10.1085/jgp.97.4.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gorman A. L., McReynolds J. S. Ionic effects on the membrane potential of hyperpolarizing photoreceptors in scallop retina. J Physiol. 1978 Feb;275:345–355. doi: 10.1113/jphysiol.1978.sp012193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
- Haynes L. W., Kay A. R., Yau K. W. Single cyclic GMP-activated channel activity in excised patches of rod outer segment membrane. Nature. 1986 May 1;321(6065):66–70. doi: 10.1038/321066a0. [DOI] [PubMed] [Google Scholar]
- Holz R., Finkelstein A. The water and nonelectrolyte permeability induced in thin lipid membranes by the polyene antibiotics nystatin and amphotericin B. J Gen Physiol. 1970 Jul;56(1):125–145. doi: 10.1085/jgp.56.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horn R., Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988 Aug;92(2):145–159. doi: 10.1085/jgp.92.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson E. C., Bacigalupo J., Vergara C., Lisman J. E. Multiple conductance states of the light-activated channel of Limulus ventral photoreceptors. Alteration of conductance state during light. J Gen Physiol. 1991 Jun;97(6):1187–1205. doi: 10.1085/jgp.97.6.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MILLER W. H. Derivatives of cilia in the distal sense cells of the retina of Pecten. J Biophys Biochem Cytol. 1958 Mar 25;4(2):227–228. doi: 10.1083/jcb.4.2.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matthews G. Single-channel recordings demonstrate that cGMP opens the light-sensitive ion channel of the rod photoreceptor. Proc Natl Acad Sci U S A. 1987 Jan;84(1):299–302. doi: 10.1073/pnas.84.1.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McReynolds J. S., Gorman A. L. Membrane conductances and spectral sensitivities of Pecten photoreceptors. J Gen Physiol. 1970 Sep;56(3):392–406. doi: 10.1085/jgp.56.3.392. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McReynolds J. S., Gorman A. L. Photoreceptor potentials of opposite polarity in the eye of the scallop, Pecten irradians. J Gen Physiol. 1970 Sep;56(3):376–391. doi: 10.1085/jgp.56.3.376. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nasi E. Electrophysiological properties of isolated photoreceptors from the eye of Lima scabra. J Gen Physiol. 1991 Jan;97(1):17–34. doi: 10.1085/jgp.97.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nasi E. Two light-dependent conductances in Lima rhabdomeric photoreceptors. J Gen Physiol. 1991 Jan;97(1):55–72. doi: 10.1085/jgp.97.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nasi E. Whole-cell clamp of dissociated photoreceptors from the eye of Lima scabra. J Gen Physiol. 1991 Jan;97(1):35–54. doi: 10.1085/jgp.97.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rae J., Cooper K., Gates P., Watsky M. Low access resistance perforated patch recordings using amphotericin B. J Neurosci Methods. 1991 Mar;37(1):15–26. doi: 10.1016/0165-0270(91)90017-t. [DOI] [PubMed] [Google Scholar]
- Ruknudin A., Song M. J., Sachs F. The ultrastructure of patch-clamped membranes: a study using high voltage electron microscopy. J Cell Biol. 1991 Jan;112(1):125–134. doi: 10.1083/jcb.112.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sokabe M., Sachs F., Jing Z. Q. Quantitative video microscopy of patch clamped membranes stress, strain, capacitance, and stretch channel activation. Biophys J. 1991 Mar;59(3):722–728. doi: 10.1016/S0006-3495(91)82285-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wong F., Knight B. W., Dodge F. A. Adapting bump model for ventral photoreceptors of Limulus. J Gen Physiol. 1982 Jun;79(6):1089–1113. doi: 10.1085/jgp.79.6.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zimmerman A. L., Baylor D. A. Cyclic GMP-sensitive conductance of retinal rods consists of aqueous pores. Nature. 1986 May 1;321(6065):70–72. doi: 10.1038/321070a0. [DOI] [PubMed] [Google Scholar]
