Skip to main content
SpringerLink
Log in

Morphological Identification of Melanopsin-Expressing Retinal Ganglion Cell Subtypes in Mice

  • Protocol
  • First Online:
Mouse Retinal Phenotyping

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1753))

Abstract

Melanopsin-expressing, intrinsically photosensitive retinal ganglion cells (ipRGCs) are a relatively recently discovered class of photoreceptor. ipRGCs can be subdivided into at least five subtypes (M1-M5), each of which has a distinct complement of morphological and physiological properties. ipRGC subtypes can be identified morphologically based on a combination of dendritic morphology and immunostaining for a cell-type specific marker. In this chapter, we describe methods for conclusively identifying each of the five ipRGC subtypes through a combination of patch clamp electrophysiology, Neurobiotin filling, visualization of ipRGC dendrites, and immunostaining for the marker SMI-32.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Schmidt TM, Chen SK, Hattar S (2011) Intrinsically photosensitive retinal ganglion cells: many subtypes, diverse functions. Trends Neurosci 34:572–580. https://doi.org/10.1016/j.tins.2011.07.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Schmidt TM, Kofuji P (2009) Functional and morphological differences among intrinsically photosensitive retinal ganglion cells. J Neurosci 29:476–482. https://doi.org/10.1523/JNEUROSCI.4117-08.2009.Functional

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Schmidt TM, Kofuji P (2010) Differential cone pathway influence on intrinsically photosensitive retinal ganglion cell subtypes. J Neurosci 30:16262–16271. https://doi.org/10.1523/JNEUROSCI.3656-10.2010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Schmidt TM, Kofuji P (2011) Structure and function of bistratified intrinsically photosensitive retinal ganglion cells in the mouse. J Comp Neurol 519:1492–1504. https://doi.org/10.1002/cne.22579

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Ecker JL, Dumitrescu ON, Wong KY et al (2010) Melanopsin-expressing retinal ganglion-cell photoreceptors: cellular diversity and role in pattern vision. Neuron 67:49–60. https://doi.org/10.1016/j.neuron.2010.05.023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Estevez ME, Fogerson PM, Ilardi MC et al (2012) Form and function of the M4 cell, an intrinsically photosensitive retinal ganglion cell type contributing to geniculocortical vision. J Neurosci 32:13608–13620. https://doi.org/10.1523/JNEUROSCI.1422-12.2012.Form

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Schmidt TM, Alam NM, Chen S et al (2014) A role for melanopsin in alpha retinal ganglion cells and contrast detection. Neuron 82:781–788. https://doi.org/10.1016/j.neuron.2014.03.022

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Lucas RJ, Hattar S, Takao M et al (2003) Diminished pupillary light reflex at high irradiances in melanopsin-knockout mice. Science 299:245–247. https://doi.org/10.1126/science.1077293

    Article  CAS  PubMed  Google Scholar 

  9. Güler AD, Ecker JL, Lall GS et al (2008) Melanopsin cells are the principal conduits for rod/cone input to non-image forming vision. Nature 453:102–105. https://doi.org/10.1038/nature06829.Melanopsin

    Article  PubMed  PubMed Central  Google Scholar 

  10. Chen S-K, Badea TC, Hattar S (2011) Photoentrainment and pupillary light reflex are mediated by distinct populations of ipRGCs. Nature 476:92–95. https://doi.org/10.1038/nature10206

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hatori M, Le H, Vollmers C et al (2008) Inducible ablation of melanopsin-expressing retinal ganglion cells reveals their central role in non-image forming visual responses. PLoS One 3(6):e2451. https://doi.org/10.1371/journal.pone.0002451

    Article  PubMed  PubMed Central  Google Scholar 

  12. Altimus CM, Guler AD, Villa KL et al (2008) Rods-cones and melanopsin detect light and dark to modulate sleep independent of image formation. Proc Natl Acad Sci 105:19998–20003. https://doi.org/10.1073/pnas.0808312105

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. LeGates TA, Altimus CM, Wang H et al (2012) Aberrant light directly impairs mood and learning through melanopsin-expressing neurons. Nature 491:594–598. https://doi.org/10.1038/nature11673

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Hu C, Hill DD, Wong KY (2013) Intrinsic physiological properties of the five types of mouse ganglion-cell photoreceptors. J Neurophysiol 109:1876–1889. https://doi.org/10.1152/jn.00579.2012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zhao X, Stafford BK, Godin AL et al (2014) Photoresponse diversity among the five types of intrinsically photosensitive retinal ganglion cells. J Physiol 592:1619–1636. https://doi.org/10.1113/jphysiol.2013.262782

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Zaghloul KA, Boahen K, Demb JB (2003) Different circuits for ON and OFF retinal ganglion cells cause different contrast sensitivities. J Neurosci 23:2645–2654. doi: 23/7/2645 [pii]

    CAS  PubMed  Google Scholar 

  17. Boycott BB, Wässle H (1974) The morphological types of ganglion cells of the domestic cat’s retina. J Physiol 240:397–419. https://doi.org/10.1002/cne.903240411

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Schmidt TM, Taniguchi K, Kofuji P (2008) Intrinsic and extrinsic light responses in melanopsin-expressing ganglion cells during mouse development. J Neurophysiol 100:371–384. https://doi.org/10.1152/jn.00062.2008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Weng S, Estevez ME, Berson DM (2013) Mouse ganglion-dell photoreceptors are driven by the most sensitive rod pathway and by both types of cones. PLoS One 8(6):e66480. https://doi.org/10.1371/journal.pone.0066480

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Vuong HE, Hardi CN, Barnes S et al (2015) Parallel inhibition of dopamine amacrine cells and intrinsically photosensitive retinal ganglion cells in a non-image-forming visual circuit of the mouse retina. J Neurosci 35:15955–15970. https://doi.org/10.1523/JNEUROSCI.3382-15.2015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Do MT, Kang SH, Xue T et al (2009) Photon capture and signalling by melanopsin retinal ganglion cells. Nature 457:281–287. https://doi.org/10.1038/nature07682

    Article  CAS  PubMed  Google Scholar 

  22. Schmidt TM, Kofuji P (2011) An isolated retinal preparation to record light response from genetically labeled retinal ganglion cells. J Vis Exp 47:e2367. https://doi.org/10.3791/2367

    Google Scholar 

  23. Boulton AA, Baker GB, Walz W (1995) Patch-clamp applications and protocols. Patch-Clamp Appl Protoc 26:259–305. https://doi.org/10.1385/0896033112

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurobiology, Northwestern University, Evanston, IL, USA

    Seul Ki Lee & Tiffany M. Schmidt

Authors
  1. Seul Ki Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tiffany M. Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tiffany M. Schmidt .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology, University Hospital Schleswig-Holstein, Kiel, Germany

    Naoyuki Tanimoto

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lee, S.K., Schmidt, T.M. (2018). Morphological Identification of Melanopsin-Expressing Retinal Ganglion Cell Subtypes in Mice. In: Tanimoto, N. (eds) Mouse Retinal Phenotyping. Methods in Molecular Biology, vol 1753. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7720-8_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7720-8_19

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7719-2

  • Online ISBN: 978-1-4939-7720-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation