Abstract
Purpose: Compelling new evidence reveals a close link between the gut microbiome and the pathogenesis of neovascular age-related macular degeneration (nAMD). Germfree (GF) animal models are the current gold standard for studying host the microbe interactionsinvivo;yet,noGFanimalmodelsofnAMDareavailabletoday.Thisprotocol describes gnotobiotic operations and assembly for a laser-induced choroidal neovascularization (CNV) model in GF mice to study the gut microbiome in neovascular AMD. Methods: We developed a step-wise approach to performing retinal laser photocoagulation in GF C57BL/6J mice that were bred and maintained at the gnotobiotic facility. Following a strict sterility protocol, we administered laser photocoagulation via an Argon 532-nm laser attached to a customized slit-lamp delivery system. Sterility was confirmed by weekly fecal cultures and reverse transcriptase–polymerase chain reaction. Results: The experiment was repeated twice at different time points using seven mice (14 eyes). Stool cultures and RT-PCR remained negative for 14 days post-procedure in all mice. Lectin immunostaining performed on choroidal flatmounts confirmed the presence of CNV lesions 2 weeks after laser treatment. Conclusions: We established a GF mouse model of nAMD with detailed guidelines to deliver retinal laser in GF mice maintaining sterility after the laser procedure. Translational Relevance: To our knowledge, this is the first protocol that describes a GF murine model of laser-induced CNV. In addition to nAMD, this animal model can be used to investigate host–microbial interactions in other eye diseases with laser-induced mouse models such as glaucoma and retinal vein occlusion.
Original language | English (US) |
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Article number | 14 |
Journal | Translational Vision Science and Technology |
Volume | 10 |
Issue number | 9 |
DOIs | |
State | Published - Aug 2021 |
Externally published | Yes |
Keywords
- age-related macular degeneration
- animal model
- choroidal neovascularization
- germ-free
- gnotobiotic
- microbiome
ASJC Scopus subject areas
- Biomedical Engineering
- Ophthalmology