July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Multimodal Longitudinal Imaging of Microglia dynamics in laser-induced acute inflammation and Choroidal Neovascularization (L-CNV)
Author Affiliations & Notes
  • Jonathan Luisi
    Pharmacology and Toxicology, University of Texas Medical Branch at Galveston, Galveston, Texas, United States
    Center for Biomedical Engineering, University of Texas Medical Branch, Galveson, Texas, United States
  • Wenbo Zhang
    Department of Opthalmology and Visual Science, University of Texas Medical Branch, Galveston, Texas, United States
  • Gracie Vargas
    Center for Biomedical Engineering, University of Texas Medical Branch, Galveson, Texas, United States
  • Massoud Motamedi
    Center for Biomedical Engineering, University of Texas Medical Branch, Galveson, Texas, United States
    Department of Opthalmology and Visual Science, University of Texas Medical Branch, Galveston, Texas, United States
  • Footnotes
    Commercial Relationships   Jonathan Luisi, None; Wenbo Zhang, None; Gracie Vargas, None; Massoud Motamedi, None
  • Footnotes
    Support  NEHS T32ES007254
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 4605. doi:
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      Jonathan Luisi, Wenbo Zhang, Gracie Vargas, Massoud Motamedi; Multimodal Longitudinal Imaging of Microglia dynamics in laser-induced acute inflammation and Choroidal Neovascularization (L-CNV). Invest. Ophthalmol. Vis. Sci. 2018;59(9):4605.

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

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Abstract

Purpose : CNV is a multi-stage neuroinflammation process with elements of vascular and immune coupling that cannot be noninvasively and longitudinally studied with traditional imaging techniques. In this study, multimodal in vivo imaging by fluorescent fundus imaging and Optical coherence tomography Angiography (OCTA) was performed for longitudinal study of microglial (MG) and vascular responses in a laser-induced model of CNV using Cx3Xr1eGFP transgenic mice.

Methods : Age-matched C57-BL6 and Cx3Cr1eGFP mice were assigned to acute or CNV groups. The L-CNV model was induced using 532 nm laser set at 180mw for 70ms with acute lesions made using 46mW of power for 200ms (Micron III). Left retinas served as controls. Imaging time points following lesion induction were 1h, and days 1, 3, 7, 14 and 21. AF imaging (Heidelberg Spectralis) was used to track MG in transgenic Cx3Cr1eGFP mice in conjuction with OCTA (Bioptigen). High-resolution OCT B-scans were provided lesion metrics and progression, while automated cell counting of fluorescent images provided MG metrics. Vascular and microglia morphology was validated using immunofluorescent imaging of retinal flatmounts.

Results : Combining metrics obtained from lesion and MG parameters we established indicators of the acute phase, transition, and chronic phases of the inflammatory response. In the case of acute lesions, MG activation occurred within 24 hours, migrated to the lesion site by D3, and returned to resting state around D14 but with an increased (+10%) density. In L-CNV the MG migration occurred D3-7, and both remain elevated (+20% density) displaying amoeboid morphology in the CNV lesion and with sub-retinal accumulation D14 through D21. Using OCTA we determined that the swelling associated with CNV lesions corresponds to a loss of inner retinal capillary perfusion from 24h to D7; reperfusion was observed D 7-14.

Conclusions : This study provided insight into vascular and immune coupling present in CNV. The distribution of MG activation patterns corresponding to acute inflammation and progression to chronic phase CNV were characterized by quantitative imaging. CNV lesions continue to progress into the chronic phase D7-21 during which MG are slower to migrate and remain activated and elevated from D14-21. With both injuries, the increased microglia cell count at 21 days indicates potentiation of the inflammatory response.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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