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Measuring the optokinetic response of zebrafish larvae


Our laboratory screens for visual mutants by examining larval eye movements in response to rotating illuminated stripes. This behavior, which is termed an optokinetic response (OKR), is a reflex that appears in zebrafish at the same time as the development of the visual system. The OKR can be accurately measured by 4 d post-fertilization, which is the age when larvae begin foraging for food. The OKR requires 1 min per larva analyzed. After identifying fish with defective eye movements, we conduct secondary screens (such as histological analysis and electroretinography) to identify the subset of fish with disruptions in the function of the outer retina. This paper describes our protocol for the OKR. Our setup is simple to construct and the materials needed are inexpensive. This makes our system especially useful for new undergraduate and graduate students, as well as introductory science lecturers.

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  1. 1

    Brockerhoff, S.E. et al. A behavioral screen for isolating zebrafish mutants with visual system defects. Proc. Natl. Acad. Sci. USA 92, 10545–10549 (1995).

  2. 2

    Clark, D.T. in Biology (Univ. of Oregon Press, Eugene, Oregon, 1981).

  3. 3

    Solnica-Krezel, L., Schier, A.F. & Driever, W. Efficient recovery of ENU-induced mutations from the zebrafish germline. Genetics 136, 1401–1420 (1994).

  4. 4

    Mullins, M.C. & Nusslein-Volhard, C. Mutational approaches to studying embryonic pattern formation in the zebrafish. Curr. Opin. Genet. Dev. 3, P648–P654 (1993).

  5. 5

    Mullins, M.C., Hammerschmidt, M., Haffter, P. & Nusslein-Volhard, C. Large-scale mutagenesis in the zebrafish: in search of genes controlling development in a vertebrate. Curr. Biol. 4, 189–202 (1994).

  6. 6

    Easter, S.S. Jr. & Nicola, G.N. The development of vision in the zebrafish (Danio rerio). Dev. Biol. 180, 646–663 (1996).

  7. 7

    Branchek, T. & Bremiller, R. The development of photoreceptors in the zebrafish, Brachydanio rerio. I. Structure. J. Comp. Neurol. 224, 107–115 (1984).

  8. 8

    Branchek, T. The development of photoreceptors in the zebrafish, Brachydanio rerio. II. Function. J. Comp. Neurol. 224, 116–122 (1984).

  9. 9

    Brockerhoff, S.E. et al. Light stimulates a transducin-independent increase of cytoplasmic Ca2+ and suppression of current in cones from the zebrafish mutant nof. J. Neurosci. 23, 470–480 (2003).

  10. 10

    Westerfield, M. The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Brachydanio rerio) (Univ. of Oregon Press, Eugene, Oregon, 1995).

  11. 11

    Muto, A. et al. Forward genetic analysis of visual behavior in zebrafish. PLoS Genet. 1, e66 (2005).

  12. 12

    Taylor, M.R., Hurley, J.B., Van Epps, H.A. & Brockerhoff, S.E. A zebrafish model for pyruvate dehydrogenase deficiency: rescue of neurological dysfunction and embryonic lethality using a ketogenic diet. Proc. Natl. Acad. Sci. USA 101, 4584–4589 (2004).

  13. 13

    Robinson, J., Schmitt, E.A., Harosi, F.I., Reece, R.J. & Dowling, J.E. Zebrafish ultraviolet visual pigment: absorption spectrum, sequence, and localization. Proc. Natl. Acad. Sci. USA 90, 6009–6012 (1993).

  14. 14

    Brockerhoff, S.E., Hurley, J.B., Niemi, G.A. & Dowling, J.E. A new form of inherited red-blindness identified in zebrafish. J. Neurosci. 20, 1–8 (1997).

  15. 15

    Rinner, O., Rick, J.M. & Neuhauss, S.C. Contrast sensitivity, spatial and temporal tuning of the larval zebrafish optokinetic response. Invest. Ophthalmol. Vis. Sci. 46, 137–142 (2005).

  16. 16

    Orger, M.B. et al. Behavioral screening assays in zebrafish. Methods Cell Biol. 77, 53–68 (2004).

  17. 17

    Van Epps, H.A. et al. The zebrafish nrc mutant reveals a role for the polyphosphoinositide phosphatase synaptojanin 1 in cone photoreceptor ribbon anchoring. J. Neurosci. 24, 8641–8650 (2004).

  18. 18

    Kay, J.N., Finger-Baier, K.C., Roeser, T., Staub, W. & Baier, H. Retinal ganglion cell genesis requires lakritz, a zebrafish atonal homolog. Neuron 30, 725–736 (2001).

  19. 19

    Wong, K.Y., Adolph, A.R. & Dowling, J.E. Retinal bipolar cell input mechanisms in giant danio. I. Electroretinographic analysis. J. Neurophysiol. 93, 84–93 (2005).

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S.E.B. is supported by National Institutes of Health grant number EY015165.

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The author declares no competing financial interests.

Supplementary information

Supplementary Video 1

Eye movements of two larvae. The OKR can be seen in the left hand larvae only. (MOV 1536 kb)

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Figure 1
Figure 2: Example of a graphical representation of 6 d post-fertilization zebrafish larva eye movements.


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