NF-kB activation as a biomarker of light injury using a transgenic mouse model

Ginger M. Pocock, Adam Boretsky, Heuy Ching Wang, Dallas Golden, Praveena Gupta, Gracie Vargas, Jeffrey W. Oliver, Massoud Motamedi

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

The spatial and temporal activation of NF-kB (p65) was monitored in the retina of a transgenic mouse model (cis-NFkB-EGFP) in vivo after receiving varying grades of laser induced thermal injury in one eye. Baseline images of the retinas from 26 mice were collected prior to injury and up to five months post-exposure using a Heidelberg Spectralis HRA confocal scanning laser ophthalmoscope (cSLO) with a spectral domain optical coherence tomographer (SDOCT). Injured and control eyes were enucleated at discrete time points following laser exposure for cryosectioning to determine localization of NF-kB dependent enhanced green fluorescent protein (EGFP) reporter gene expression within the retina using fluorescence microscopy. In addition, EGFP basal expression in brain and retinal tissue from the cis-NFkB-EGFP was characterized using two-photon imaging. Regions of the retina exposed to threshold and supra-threshold laser damage evaluated using fluorescence cSLO showed increased EGFP fluorescence localized to the exposed region for a duration that was dependent upon the degree of injury. Fluorescence microscopy of threshold damage revealed EGFP localized to the outer nuclear region and retinal pigment epithelial layer. Basal expression of EGFP imaged using two-photon microscopy was heterogeneously distributed throughout brain tissue and confined to the inner retina. Results show cis-NF-kB-EGFP reporter mouse can be used for in vivo studies of light induced injury to the retina and possibly brain injury.

Original languageEnglish (US)
Title of host publicationOptical Interactions with Tissue and Cells XXIII
DOIs
StatePublished - 2012
EventOptical Interactions with Tissue and Cells XXIII - San Francisco, CA, United States
Duration: Jan 23 2012Jan 25 2012

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume8221
ISSN (Print)1605-7422

Other

OtherOptical Interactions with Tissue and Cells XXIII
Country/TerritoryUnited States
CitySan Francisco, CA
Period1/23/121/25/12

Keywords

  • NF-kB
  • brain
  • confocal scanning laser ophthalmoscope
  • fluorescence microscopy
  • inflammation
  • laser
  • laser damage
  • mouse
  • retina
  • spectral domain optical coherence tomography
  • transgenic
  • two-photon imaging

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Biomaterials

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