In Vivo Multimodal Imaging of NF-kappaB (pB) Spatial and Temporal Activation Following Light Injury in the Mouse Retina
Abstract
BALB/c and C57BL/6 wild type mice and (cis-NF- B-EGFP) transgenic mouse model were used to perform in vivo experiments of laserinduced thermal damage to the retina. A Heidelberg Spectralis HRA confocal scanning laser ophthalmoscope with a spectral domain optical coherence tomographer was used to obtain fundus and cross-sectional images of laser induced injury in the retina. Sub-threshold, threshold, and supra-threshold lesions were imaged up to 10 days post exposure using optical coherence tomography (OCT), infrared reflectance, red-free reflectance, fluorescence angiography, and autofluorescence imaging modalities. The spatial and temporal activation of Nuclear factor-kappa B (NF- B) (p65) was monitored in the retina of the cis-NF- B-EGFP mouse. In addition, EGFP basal expression in brain and retinal tissue from the cis-NF- B-EGFP was characterized using two-photon imaging. Injured and control eyes were enucleated at discrete time points following laser exposure for cryosectioning and confocal microscopy to determine localization of NF- B dependent enhanced green fluorescent protein (EGFP) reporter gene expression within the retina. Furthermore, immunohistochemistry for GFAP, IBA-1, NeuN, CD65, and glutamine synthetase were used to examine the retinal structures and cell population expressing upregulated activation of p65 as a result of laser induced injury.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 15, 2014
- Accession Number
- ADA595708
Entities
People
- Adam Boretsky
- Brent J. Lavey
- Ginger M. Pocock
- Gracie Vargas
- Heuy-ching H. Wang
- Massoud Motamedi
- Praveena Gupta
- Wenbo Zhang
Organizations
- Air Force Research Laboratory