April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Comprehensive Analysis Of The Central Projections Of Melanopsin-expressing Retinal Ganglion Cell In Mice
Author Affiliations & Notes
  • Satchidananda Panda
    Regulatory Biolgy, Salk Institute, San Diego, California
  • Sheena R. Keding
    Regulatory Biolgy, Salk Institute, San Diego, California
  • Hiep D. Le
    Regulatory Biolgy, Salk Institute, San Diego, California
  • Daniel Gibbs
    Regulatory Biolgy, Salk Institute, San Diego, California
  • James Fitzpatrick
    Regulatory Biolgy, Salk Institute, San Diego, California
  • Megumi Hatori
    Regulatory Biolgy, Salk Institute, San Diego, California
  • Footnotes
    Commercial Relationships  Satchidananda Panda, None; Sheena R. Keding, None; Hiep D. Le, None; Daniel Gibbs, None; James Fitzpatrick, None; Megumi Hatori, None
  • Footnotes
    Support  NIH grant EY016807, JSPS fellowship
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3460. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Satchidananda Panda, Sheena R. Keding, Hiep D. Le, Daniel Gibbs, James Fitzpatrick, Megumi Hatori; Comprehensive Analysis Of The Central Projections Of Melanopsin-expressing Retinal Ganglion Cell In Mice. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3460.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : Melanopsin (Opn4) and melanopsin expressing retinal ganglion cells (mRGCs) have now been implicated in several photoresponses. These mRGCs project their axons to the suprachiasmatic nucleus (SCN) and a few other brain regions that directly or indirectly regulate all non-image forming visual processes including circadian photoentrainment, pupil constriction, pineal melatonin regulation and light regulation of activity/rest. Identifying the full complement of mRGCs and their central projection is critical to understanding the cellular basis of melanopsin function. However, the existing methods to mark melanopsin cells and map their projections are insufficient. We have developed novel methods to comprehensively map mRGC projections.

Methods: : We bred Opn4Cre mouse to Z/EG or Z/AP mice, which allows Cre-dependent expression of green fluorescent protein (GFP) or human alkaline phosphatase (AP) from a β-actin promoter. We also used Adeno-associated virus serotype 2 (AAV2) expressing a green or red fluorescent protein in Cre-dependent manner in intravitreal injections to Opn4Cre mice to specifically label the mRGCs. Flat mount retina, retina sections, and brain sections were stained for the respective reporters.

Results: : In the retina of Opn4Cre/+;Z/EG mice, GFP expressing cells were mostly found in the retinal ganglion cell (RGC) sub-layer, and these cells had extensive dendritic arborization characteristic of the mRGCs. In Opn4Cre/+;Z/AP mice, the mRGCs strongly innervate the SCN. Additionally, mRGCs axon termini also sparsely innervate various other hypothalamic regions. Surprisingly, the AP staining also revealed extensive projection of the mRGCs in the lateral geniculate complex which is involved in image-forming vision. However, in adult mice, it was reported that there was a chance of insufficient labeling of Z/EG and Z/AP lines probably due to the methylation of the β -actin promoter used for these lines. Therefore we had intravitreally injected AAV2 viruses that express mCherry or EGFP from a different promoter in Cre-dependent manner to adult mice. This attempt resulted in reproducible success in transducing vast majority of mRGCs only in the injected retina than the retina of Opn4Cre/+;Z/EG mice.

Conclusions: : We found that the projection patterns of mRGCs were much more extensive than previously reported. We also developed the novel viral strategy to comprehensively map mRGC projections. Such strategy can now be applied to any mouse expressing Cre in the retina.

Keywords: ganglion cells • circadian rhythms • photoreceptors 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×