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

doi:10.1117/12.908835

9 February 2012 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

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Proceedings Volume 8221, Optical Interactions with Tissue and Cells XXIII; 822108 (2012) https://doi.org/10.1117/12.908835
Event: SPIE BiOS, 2012, San Francisco, California, United States

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.

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Ginger M. Pocock, Adam Boretsky, Heuy-Ching Wang, Dallas Golden, Praveena Gupta, Gracie Vargas, Jeffrey W Oliver, and Massoud Motamedi "NF-kB activation as a biomarker of light injury using a transgenic mouse model", Proc. SPIE 8221, Optical Interactions with Tissue and Cells XXIII, 822108 (9 February 2012); https://doi.org/10.1117/12.908835

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KEYWORDS

Retina

Luminescence

Injuries

Brain

Microscopy

Neuroimaging

Confocal microscopy

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