Hostname: page-component-8448b6f56d-c47g7 Total loading time: 0 Render date: 2024-04-24T00:43:48.691Z Has data issue: false hasContentIssue false

An enzymatically enhanced recording technique for Limulus ventral photoreceptors: Physiology, biochemistry, and morphology

Published online by Cambridge University Press:  02 June 2009

Hui-Juan Zhang
Affiliation:
Department of Zoology, University of Maine, Orono
Robert N. Jinks
Affiliation:
Institute for Sensory Research, Syracuse University, Syracuse Department of Bioengineering and Neuroscience, Syracuse University, Syracuse
Anne C. Wishart
Affiliation:
The Whitney Laboratory, University of Florida, St. Augustine
Barbara-Anne Battelle
Affiliation:
The Whitney Laboratory, University of Florida, St. Augustine
Steven C. Chamberlain
Affiliation:
Institute for Sensory Research, Syracuse University, Syracuse Department of Bioengineering and Neuroscience, Syracuse University, Syracuse
Wolf H. Fahrenbach
Affiliation:
Laboratory of Electron Microscopy, Oregon Regional Primate Research Center, Beaverton
Leonard Kass
Affiliation:
Department of Zoology, University of Maine, Orono

Abstract

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.

Type
Research Articles
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anderson, P.A.V. (1987). Properties and pharmacology of a TTX-insensitive Na+ current in neurons of the jellyfish Cyanea capillata. Journal of Experimental Biology 133, 231248.Google Scholar
Bacigalupo, J. & Johnson, E.C. (1992). Localization of phototransduction in Limulus ventral photoreceptors: A demonstration using cell-free rhabdomeric vesicles. Visual Neuroscience 8, 4147.Google Scholar
Bacigalupo, J. & Lisman, J.E. (1983). Single-channel currents activated by light in Limulus ventral photoreceptors. Nature 304, 268270.CrossRefGoogle ScholarPubMed
Barlow, R.B. Jr & Kaplan, E. (1971). Limulus lateral eye: Properties of receptor units in the unexcised eye. Science 174, 10271029.Google Scholar
Barlow, R.B. Jr & Kaplan, E. (1977). Properties of visual cells in the lateral eye of Limulus in situ: Intracellular recordings. Journal of General Physiology 69, 203220.CrossRefGoogle ScholarPubMed
Barlow, R.B. Jr, Bolanowski, S.J. Jr & Brachman, M.L. (1977). Efferent optic nerve fibers mediate circadian rhythms in the Limulus eye. Science 197, 8689.Google Scholar
Barlow, R.B. Jr, Kaplan, E., Renninger, G.H., & Saito, T. (1985). Efferent control of circadian rhythms in Limulus lateral eye. Neuroscience Research (Suppl.) 2, S65–S78.Google Scholar
Barlow, R.B. Jr, Kaplan, E., Renninger, G.H., & Saito, T. (1987). Circadian rhythms in Limulus photoreceptors: I. Intracellular studies. Journal of General Physiology 89, 353378.CrossRefGoogle ScholarPubMed
Batra, R. & Barlow, R.B. Jr (1990). Circadian rhythms in temporal response of the Limulus lateral eye. Journal of General Physiology 95, 229244.Google Scholar
Battelle, B-A. & Evans, J.A. (1984). Octopamine release from centrifugal fibers of the Limulus visual system. Journal of Neurochemistry 46, 14641472.CrossRefGoogle Scholar
Battelle, B-A., Evans, J.A., & Chamberlain, S.C. (1982). Efferent fibers to Limulus eyes synthesize and release octopamine. Science 216, 12501252.CrossRefGoogle Scholar
Bayer, D.S., & Barlow, R.B. Jr (1978). Limulus ventral eye: Physiological properties of photoreceptor cells in organ culture medium. Journal of General Physiology 72, 539564.Google Scholar
Berry, M.N., & Friend, D.S. (1969). High-yield preparation of isolated rat-liver parenchymal cells. Journal of Cell Biology 43, 506520.Google Scholar
Calman, B.G., & Chamberlain, S.C. (1982). Distinct lobes of Limulus ventral photoreceptors. II. Structure and ultrastructure. Journal of General Physiology 80, 839862.Google Scholar
Chamberlain, S.C., (1984). Are mitochondria the internal calcium compartment for light adaptation in Limulus photoreceptors? Investigative Ophthalmology and Visual Science (Suppl.) 26, 64.Google Scholar
Clark, A.W., Millecchia, R. & Mauro, A. (1969). The ventral photoreceptor cell of Limulus. I. The microanatomy. Journal of General Physiology 54, 289309.Google Scholar
Edwards, S.C., Andrews, A.W., Renninger, G.H., Wiebe, E.M., & Battelle, B-A. (1990). Efferent innervation of Limulus eyes in vitro phosphorylates a 122 kD protein. Biological Bulletin 178, 267268.Google Scholar
Edwards, S.C., & Battelle, B-A. (1987). Octopamine- and cAMP-stimulated phosphorylation of a protein in Limulus ventral and lateral eye. Journal of Neuroscience 7, 28112820.Google Scholar
Fahrenbach, W.H., (1969). The morphology of the eyes of Limulus. II. Ommatidia of the compound eye. Zeitschrift für Zellforschung und Mikroskopische Anatomie 93, 451483.Google Scholar
Hanna, B., Jinks, R.N., Zhang, H.-j., Kass, L., Renninger, G.H., & Chamberlain, S.C. (1991). Ultrastructure of Limulus photoreceptors in experimental protocols: Physiologists’ favorite recipes. Society for Neuroscience Abstracts 17, 298.Google Scholar
Hanna, W.J.B., Johnson, E.C., Chaves, D. & Renninger, G.H. (1993). Photoreceptor cells dissociated from the compound lateral eye of the horseshoe crab, Limulus polyphemus. II: Function. Visual Neuroscience 10, 609620.Google Scholar
Harper, E. (1980). Collagenase. Annual Review of Biochemistry 49, 10631078.CrossRefGoogle Scholar
Herman, K.G., (1991 a). Two classes of Limulus ventral photoreceptors. Journal of Comparative Neurology 303, 110.CrossRefGoogle ScholarPubMed
Herman, K.G., (1991 b). Light-stimulated rhabdom turnover in Limulus ventral photoreceptors maintained in vitro. Journal of Comparative Neurology 303, 1121.Google Scholar
Jinks, R.N., & Chamberlain, S.C. (1990). Does seawater cause structural light adaptation in Limulus ventral photoreceptors in vitro? – A study of organ culture vs. seawater incubation. Society for Neuroscience Abstracts 16, 406.Google Scholar
Jinks, R.N., Hanna, W.J.B., Renninger, G.H. & Chamberlain, S.C. (1993). Photoreceptor cells dissociated from the compound lateral eye of the horseshoe crab, Limulus polyphemus. I: Structure and ultrastructure. Visual Neuroscience 10, 597607.CrossRefGoogle ScholarPubMed
Kaplan, E. & Barlow, R.B. Jr (1975). Properties of visual cells in the lateral eye of Limulus in situ: Extracellular recordings. Journal of General Physiology 66, 303326.Google Scholar
Kaplan, E. & Barlow, R.B. Jr (1980). Circadian clock in Limulus brain increases response and decreases noise of retinal photoreceptors. Nature 286, 393395.CrossRefGoogle ScholarPubMed
Kaplan, E., Barlow, R.B. Jr., Renninger, G. & Purpura, K. (1990). Circadian rhythms in Limulus photoreceptors. II. Quantum bumps. Journal of General Physiology 96, 665685.Google Scholar
Kass, L., Pelletier, J.L., Renninger, G.H. & Barlow, R.B. Jr (1988). Efferent neurotransmission of circadian rhythms in Limulus lateral eye. II. Intracellular recordings in vitro. Journal of Comparative Physiology A 164, 95105.CrossRefGoogle ScholarPubMed
Kass, L. & Renninger, G.H. (1988). Circadian changes in function of Limulus ventral photoreceptors. Visual Neuroscience 1, 311.Google Scholar
Kass, L., Renninger, G.H., Zhang, H.-J. & Pelletier, J.L. (1990). Whole-cell recordings from Limulus ventral photoreceptors. Investigative Ophthalmology and Visual Science (Suppl.) 31, 389.Google Scholar
Kass, L., Pelletier, J.L. & Zhang, H.J. (1991). Circadian clock modulates light-activated conductances and quantal bumps in Limulus ventral photoreceptors. Investigative Ophthalmology and Visual Science (Suppl.) 32, 672.Google Scholar
Kass, L. & Zhang, H.-J. (1992). Clock controls gain in Limulus photoreceptor by changing voltage-dependent conductances. Investigative Ophthalmology and Visual Science (Suppl.) 33, 1327.Google Scholar
Kruse, P.F., & Patterson, M.K. (1972). Tissue Culture, Methods and Applications. New York: Academic Press.Google Scholar
Lisman, J.E., & Brown, J.E. (1971). Two light-induced processes in the photoreceptor cells of Limulus ventral eye. Journal of General Physiology 58, 544561.Google Scholar
Millecchia, R., Bradbury, J. & Mauro, A. (1966). Simple photoreceptors in Limulus polyphemus. Science 154, 11991201.Google Scholar
Millecchia, R. & Mauro, A. (1969 a). The ventral photoreceptor cell of Limulus. II. The basic photoresponse. Journal of General Physiology 54, 310330.Google Scholar
Millecchia, R. & Mauro, A. (1969 b). The ventral photoreceptor cell of Limulus. III. A voltage clamp study. Journal of General Physiology 54, 331351.CrossRefGoogle Scholar
Payne, R. & Fein, A. (1983). Localized adaptation within the rhabdomeral lobe of ventral photoreceptors. Journal of General Physiology 81, 767769.CrossRefGoogle ScholarPubMed
Renninger, G.H., (1990). Photoreceptor cells dissociated from the Limulus lateral eye. Investigative Ophthalmology and Visual Science (Suppl.) 31, 389.Google Scholar
Renninger, G.H., Kass, L., Pelletier, J.L. & Schimmel, R. (1988). The eccentric cell of the Limulus lateral eye: Encoder of circadian changes in visual responses. Journal of Comparative Physiology A 163, 259270.Google Scholar
Renninger, G.H., Schimmel, R. & Farrell, C.A. (1989). Octopamine modulates photoreceptor function in the Limulus lateral eye. Visual Neuroscience 3, 8394.Google Scholar
Seamon, K.B., Padgett, W. & Daly, J.W. (1981). Forskolin: Unique diterpene activator of adenylate cyclase in membranes and in intact cells. Proceedings of the National Academy of Sciences of the U.S.A. 78, 33633367.CrossRefGoogle ScholarPubMed
Simcox, S.A., Ryan, T.D., Tomak, P.R. & Chamberlain, S.C. (1988). A círcadian rhythm in the distribution of photoreceptor mitochondria in Limulus. Investigative Ophthalmology and Visual Science (Suppl.) 29, 350.Google Scholar
Stern, J., Chinn, K., Bacigalupo, J. & Lisman, J. (1982). Distinct lobes of Limulus ventral photoreceptors. I. Functional and anatomical properties of lobes revealed by removal of glial cells. Journal of General Physiology 80, 825837.CrossRefGoogle ScholarPubMed
Stoner, C.D. & Sirak, H.D. (1969). Osmotically-induced alterations in volume and ultrastructure of mitochondria isolated from rat liver and bovine heart. Journal of Cell Biology 43, 521538.Google Scholar
Warren, M.K., & Pierce, S.K. (1982). Two cell volume regulatory systems in the Limulus myocardium: An interaction of ions and quaternary ammonium compounds. Biological Bulletin 163, 504516.Google Scholar
Yeandle, S. & Spiegler, J.B. (1973). Light-evoked and spontaneous discrete waves in the ventral nerve photoreceptor of Limulus. Journal of General Physiology 61, 552571.Google Scholar
Zhang, H.-I., Kass, L., Pelletier, J.L., & Renninger, G.H. (1990). Modulation of voltage-dependent conductances in Limulus ventral photoreceptors by octopamine and forskolin. Investigative Ophthalmology and Visual Science (Suppl.) 31, 389.Google Scholar