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Patch-Clamp Electrophysiological Analysis of Murine Melanopsin Neurons

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Circadian Clocks

Part of the book series: Neuromethods ((NM,volume 186))

Abstract

Intrinsically photosensitive retinal ganglion cells (ipRGCs) regulate diverse aspects of mammalian physiology, including the synchronization of circadian rhythms with local time. These neurons sense light with a receptor called melanopsin and provide practically all retinal innervation of the suprachiasmatic nucleus, the master clock. Their activity is especially important over long timescales. How do ipRGCs respond to light, what signals do they send downstream, and to what extent are these signals tailored to circadian photoregulation? This chapter provides practical guidance for studying ipRGC signals in the ex vivo mouse retina using patch-clamp electrophysiology and optical stimulation. Somatic and axonal recording are covered, as are methods that include loose, cell attached, whole cell, and perforated patch. Also discussed are how particular features of the ipRGC light response affect the design and interpretation of experiments. These approaches may be useful in the broader effort to understand how a neuron’s functional properties align with its role in the organism.

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Acknowledgments

Experiments performed by the author as a postdoctoral fellow are shown with the kind permission of his advisor, King-Wai Yau. Alan Emanuel, Elliott Milner, and Andreas Liu also obtained unpublished data from their archives. Discussions with Bruce Bean enriched this chapter. Insight on the fastidiousness of pioneering retinal physiologists was provided by Solange Brown. Comments on the manuscript were given by Chinfei Chen, Franklin Caval-Holme, Hannah Blume, and Bruce Bean. Funding was provided by the National Institutes of Health (EY023648, EY025555, and EY032731).

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  1. F. M. Kirby Neurobiology Center of Boston Children’s Hospital and Harvard Medical School, Boston, MA, USA

    Michael Tri Hoang Do

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  1. Michael Tri Hoang Do
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Correspondence to Michael Tri Hoang Do .

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  1. Institute of Transformative Bio-Molecules, Nagoya University, Nagoya, Aichi, Japan

    Tsuyoshi Hirota

  2. Institute of Transformative Bio-Molecules, Nagoya University, Nagoya, Aichi, Japan

    Megumi Hatori

  3. Regulatory Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA

    Satchidananda Panda

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Do, M.T.H. (2022). Patch-Clamp Electrophysiological Analysis of Murine Melanopsin Neurons. In: Hirota, T., Hatori, M., Panda, S. (eds) Circadian Clocks. Neuromethods, vol 186. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2577-4_6

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  • DOI: https://doi.org/10.1007/978-1-0716-2577-4_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2576-7

  • Online ISBN: 978-1-0716-2577-4

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