June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Electron microscopy of human melanopsin retinal ganglion cells
Author Affiliations & Notes
  • Jesse Gale
    Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, Capital and Coast District Health Board, Wellington, New Zealand
  • Fred N Ross-Cisneros
    Doheny Eye Institute, Los Angeles, CA
  • Chiara La Morgia
    Neurological Sciences, University of Bologna, Bologna, Italy
  • Ernesto Barron
    Doheny Eye Institute, Los Angeles, CA
  • Jens Hannibal
    Clinical Biochemistry, Bispebjerg Hospital, Copenhagen, Denmark
  • Carla Giordano
    Radiology, Oncology and Pathology, Sapienza University of Rome, Rome, Italy
  • Valerio Carelli
    Neurological Sciences, University of Bologna, Bologna, Italy
  • Alfredo A Sadun
    Doheny Eye Institute, Los Angeles, CA
    Ophthalmology, UCLA, Los Angeles, CA
  • Footnotes
    Commercial Relationships Jesse Gale, None; Fred Ross-Cisneros, None; Chiara La Morgia, None; Ernesto Barron, None; Jens Hannibal, None; Carla Giordano, None; Valerio Carelli, None; Alfredo Sadun, None
  • Footnotes
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Investigative Ophthalmology & Visual Science June 2015, Vol.56, 834. doi:
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      Jesse Gale, Fred N Ross-Cisneros, Chiara La Morgia, Ernesto Barron, Jens Hannibal, Carla Giordano, Valerio Carelli, Alfredo A Sadun; Electron microscopy of human melanopsin retinal ganglion cells. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):834.

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

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Abstract

Purpose: Melanopsin-expressing retinal ganglion cells (mRGC) subserve a variety of functions, particularly pupil light responses and circadian photoentrainment, and are peculiar in their resistance to neurodegeneration in mitochondrial optic neuropathies. We here present the first ultrastructural images of a human mRGC.

Methods: Serial retinal sections from a normal 24-year-old male control were immuno-stained for melanopsin (rabbit polyclonal antibody) and mitochondrial antigens (mouse monoclonal antibody against mitochondrial extract) using diaminobenzidine (DAB). One example of an mRGC was then processed into plastic for ultrathin sectioning for transmission electron microscopy. The subtype of this mRGC could not be indentified due to twisting of the retinal section during antigen retrieval.

Results: Melanopsin was identified with electron-dense DAB immunostaining, particularly in cell processes and along cell membranes, and some DAB was seen within the cell, associated with membrane-bound structures presumed to be endoplasmic reticulum, Golgi and vesicles. The mRGC was large, with subjectively more electron lucent and vesicular cytoplasm than other ganglion cells and surrounding glia. Antigen retrieval (with heat and citrate buffer) limited the resolution of small ultrastructural features, so the mitochondrial phenotype was not characterized. Immunostaining for melanopsin and mitochondrial antigens for light microscopy has indicated high mitochondrial density in mRGCs.

Conclusions: These first electron micrographs of a human mRGC resemble previous findings in the mouse [Belenky et al. J Comp Neurol 2003;460:380-93]. Our attempt to describe the structural mitochondrial phenotype was limited by the trade off between antigen retrieval and ultrastructural preservation. Immunostaining for mitochondria at light microscopy suggests an abundant population of these organelles in the large cytoplasm of mRGCs, which may help to explain their resistance to death after injury.

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