Nicotinic acetylcholine receptor subunits in rhesus monkey retina

Invest Ophthalmol Vis Sci. 2009 Mar;50(3):1408-15. doi: 10.1167/iovs.08-2398. Epub 2008 Oct 24.

Abstract

Purpose: The purpose of this study was to detect and establish the cellular localizations of nicotinic acetylcholine receptor (nAChR) subunits in Rhesus monkey retina.

Methods: Retinas were dissected from the eyes of monkeys killed after unrelated experiments. RNA was extracted and analyzed by RT-PCR, using primers designed against human sequences of alpha3-alpha7, alpha9, and beta2-beta4 nAChR subunits. The RT-PCR products were separated by gel electrophoresis and sequenced. Frozen sections of postmortem fixed monkey eyes were immunolabeled with well-characterized and specific monoclonal antibodies against the alpha3, alpha4, alpha6, alpha7, beta2, or beta4 nAChR subunits and visualized with fluorescence labeling.

Results: Products of the predicted size for the alpha3-alpha7, alpha9, and beta2-beta4 nAChR subunits were detected by RT-PCR in Rhesus monkey retina. Homology between transcripts from monkey retina and human nucleotide sequences ranged from 93 to 99%. Immunohistochemical studies demonstrated that neurons in various cell layers of monkey retina expressed alpha3, alpha4, alpha7, or beta2 nAChR subunits and cells with the morphology of microglia were immunoreactive for the alpha6 or beta4 nAChR subunits.

Conclusions: nAChR subunits are expressed in the monkey retina and localize to diverse retinal neurons as well as putative microglia. Besides mediating visual processing, retinal nAChRs may influence refractive development and ocular pathologies such as neovascularization.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique, Indirect
  • Macaca mulatta
  • Microscopy, Fluorescence
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • RNA, Messenger / metabolism
  • Receptors, Nicotinic / genetics
  • Receptors, Nicotinic / metabolism*
  • Retina / metabolism*
  • Retinal Neurons / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA Primers
  • Protein Subunits
  • RNA, Messenger
  • Receptors, Nicotinic