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The effects of nicotine on cone and rod b-wave responses in larval zebrafish



Acetylcholine is present in and released from starburst amacrine cells in the inner plexiform layer (IPL), but its role in retinal function except, perhaps, in early development, is unclear. Nicotinic acetylcholine receptors are thought to be present on ganglion, amacrine, and bipolar cell processes in the IPL, and it is known that acetylcholine increases the spontaneous and light-evoked responses of retinal ganglion cells. The effects of acetylcholine on bipolar cells are not known, and here we report the effects of nicotine on the b-wave of the electroretinogram in larval zebrafish. The b-wave originates mainly from ON-bipolar cells, and the larval zebrafish retina is cone-dominated. Only small rod responses can be elicited with dim lights in wild-type larval zebrafish retinas, but rod responses can be recorded over a range of intensities in a mutant (no optokinetic response f) fish that has no cone function. We find that nicotine strongly enhances cone-driven b-wave response amplitudes but depresses rod driven b-wave response amplitudes without, however, affecting rod- or cone-driven b-wave light sensitivity.


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*Address correspondence to: John E. Dowling, Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, E-mail:


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Arenzana, F.J., Clemente, D., Sanchez-Gonzalez, R., Porteros, A., Aijon, J. & Arevalo, R. (2005). Development of the cholinergic system in the brain and retina of the zebrafish. Brain Research Bulletin 66, 421425.
Ariel, M. & Daw, N.W. (1982). Pharmacological analysis of directionally sensitive rabbit retinal ganglion cells. Journal of Physiology 324, 161185.
Brockerhoff, S.E., Rieke, F., Matthews, H.R., Taylor, M.R., Kennedy, B., Ankoudinova, I., Niemi, G.A., Tucker, C.L., Xiao, M., Cilluffo, M.C., Fain, G.L., & Hurley, J.B. (2003). Light stimulates a transducin-independent increase of cytoplasmic Ca2+ and suppression of current in cones from the zebrafish mutant nof. Journal of Neuroscience 23, 470480.
Caldwell, J.H., Daw, N.W. & Wyatt, H.J. (1978). Effects of picrotoxin and strychnine on rabbit retinal ganglion cells: Lateral interactions for cells with more complex receptive fields. Journal of Physiology 276, 277298.
Dmitrieva, N., Strang, C.E. & Keyser, K.T. (2007). Expression of alpha 7 nicotinic acetylcholine receptors by bipolar, amacrine, and ganglion cells of the rabbit retina. The Journal of Histochemistry and Cytochemistry 55, 461476.
Dowling, J.E. (2012). The Retina: An Approachable Part of the Brain-Revised Edition. Cambridge, MA: The Belknap Press of Harvard University Press.
Emran, F., Rihel, J., Adolph, A.R., Wong, K.Y., Kraves, S. & Dowling, J.E. (2007). OFF ganglion cells cannot drive the optokinetic reflex in zebrafish. Proceedings of the National Academy of Sciences of the United States of America 104, 1912619131.
Emran, F., Rihel, J., Adolph, A.R. & Dowling, J.E. (2010). Zebrafish larvae lose vision at night. Proceedings of the National Academy of Sciences of the United States of America 107, 60346039.
Feller, M.B., Wellis, D.P., Stellwagen, D., Werblin, F.S. & Shatz, C.J. (1996). Requirement for cholinergic synaptic transmission in the propagation of spontaneous retinal waves. Science 272, 11821187.
Feller, M.B., Butts, D.A., Aaron, H.L., Rokhsar, D.S. & Shatz, C.J. (1997). Dynamic processes shape spatiotemporal properties of retinal waves. Neuron 19, 293306.
Fried, S., Munch, T.A. & Werblin, F.S. (2002). Mechanisms and circuitry underlying directional selectivity in the retina. Nature 420, 411414.
Jurklies, B., Kaelin-Lang, A. & Niemeyer, G. (1996). Cholinergic effects on cat retina in vitro: Changes in rod- and cone-driven b-wave and optic nerve response. Vision Research 36, 797816.
Keyser, K.T., MacNeil, M.A., Dmitrieva, N., Wang, F., Masland, R.H. & Lindstrom, J.M. (2000). Amacrine, ganglion and displaced amacrine cells in the rabbit retina express nicotine acetylcholine receptors. Visual Neuroscience 17, 243752.
Li, Y.N., Tsujimura, T., Kawamura, S. & Dowling, J.E. (2012). Bipolar cell-photoreceptor connectivity in the zebrafish (Danio rerio) retina. The Journal of Comparative Neurology 520, 37863802.
Liu, J., McGlinn, A.M., Fernandes, A., Milam, A.H., Strang, C.E., Andison, M.E., Lindstrom, J.M., Keyser, K.T. & Stone, R.A. (2009). Nicotine acetylcholine receptor subunits in rhesus monkey retina. Investigative Ophthalmology & Visual Science 50, 14081415.
Masland, R.H. & Ames, A. III (1976). Responses to acetylcholine of ganglion cells in an isolated mammalian retina. Journal of Neurophysiology 39, 12201235.
Masland, R.H. & Mills, J.W. (1979). Autoradiographic identification of acetylcholine in the rabbit retina. The Journal of Cell Biology 83, 159178.
Masland, R.H., Mills, J.W. & Hayden, S.A. (1984). Acetycholine-synthesizing amacrine cells: Identification and selective staining using radioautography and fluorescent markers. Proceedings of the Royal Society of London. Series B, Biological Sciences 223, 79100.
McArdle, C.B., Dowling, J.E. & Masland, R.H. (1997). Development of outer segments and synapses in the rabbit retina. The Journal of Comparative Neurology 175, 253273.
O’Malley, D.M., Sandell, J.H. & Masland, R.H. (1992). Co-release of acetylcholine and GABA by the starburst amacrine cells. Journal of Neuroscience 12, 13941408.
Pang, J., Gao, F., Lem, J., Bramblett, D., Paul, D.L. & Wu, S.M. (2010) Direct rod input to cone BCs challenge the traditional view of mammalian BC circuitry. Proceedings of the National Academy of Sciences of the United States of America 107, 395400.
Robson, J.G. & Frishman, L.J. (1999). Dissecting the dark-adapted electroretinogram. Documenta Ophthalmologica 95, 187215.
Shatz, C.J. & Stryker, M.P. (1988). Prenatal tetrodotoxin infusion blocks segregation of retinogeniculate afferents. Science 24, 8789.
Varghese, S.B., Reid, J.C., Hartmann, E.E. & Keyser, K.T. (2011). The effects of nicotine on the human electroretinogram. Investigative Ophthalmology & Visual Science 52, 94459451.
Westerfield, M. (2000). The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish. Eugene, OR: University of Oregon Press.
Wong, K.Y., Gray, J., Hayward, C.J.C., Adolph, A.R. & Dowling, J.E. (2004). Glutamatergic mechanisms in the outer retina of larval zebrafish: Analysis of electroretinogram b- and d-waves using a novel preparation. Zebrafish 1, 121131.
Yamada, E.S., Dmitrieva, N., Keyser, K.T., Lindstrom, J.M., Hersh, L.B. & Marshak, D.W. (2003). Synaptic connections of starburst amacrine cells and localization of acetylcholine receptors in primate retinas. The Journal of Comparative Neurology 461, 7690.
Yazulla, S. & Studholme, K.M. (2001). Neurochemical anatomy of the zebrafish retina as determined by immunocytochemistry. Journal of Neurocytology 30, 551592.
Yoshida, K., Watanabe, D., Ishikane, H., Tachibana, M., Pastan, I. & Nakanishi, S. (2001). A key role of starburst amacrine cells in originating retinal directional selectivity and optokinetic eye movement. Neuron 30, 771780.
Zucker, C. & Yazulla, S. (1982). Localization of synaptic and nonsynaptic nicotine-acetylcholine receptors in the goldfish retina. The Journal of Comparative Neurology 204, 188195.


The effects of nicotine on cone and rod b-wave responses in larval zebrafish



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