Summary: The choroidal neovascularization model is one of the most popular preclinical models to study pathologic neovascularization in vivo.
In 1989, Dobi and colleagues were first to describe a rat choroidal neovascularization model (CNV), which was induced using a krypton laser. Later on, the same induction of choroidal neovascularization was successfully repeated in mice.
Rapid development of different in vivo imaging modalities turned this model into a powerful and validated tool for the screening of novel drug candidates with anti-angiogenic and/or anti-fibrotic properties. This model mimics multiple pathological features of exudative age-related macular degeneration (AMD).
The typical follow-up time is 2 weeks both in mice and in rats. The induction is unilateral leaving the contralateral eye as naïve control.
Intravitreal or systemic (IP) administration of clinically approved Aflibercept successfully can prevent vascular leak up to 7 days after the administration (Kaja et al., 2019).
|Animal species||Mice, Rats|
|Method of induction||Diode laser|
|Follow-up period||Typically 7-14 days|
|Route of compound administration||Topical (e.g. eye drops), intravitreal or subretinal injections, systemic|
|Read-outs||1. In vivo imaging|
– Fluorescein angiography (FAG)
– Spectral domain-ocular coherence tomography (SD-OCT)
2. Morphological assessment (histology, immunohistochemistry, electron microscopy, stereology)
Outcomes and Read-Outs
In vivo imaging
- Fluorescein angiography (Heidelberg Spectralis systems, Heidelberg Engineering) is used to monitor clinically relevant vascular leak longitudinally throughout the whole follow-up period (Ragauskas et al., 2018).
- SD-OCT imaging (Envisu R2210 and Envisu R2210 systems, Bioptigen Inc./Leica Microsystems) provides confirmation of Bruchs’ membrane damage (successful induction), cross-sectional view of pathological features, and enables quantitative evaluation of the lesion (Ragauskas et al., 2018).
- Retinal and choroidal flatmounts serve for secondary validation of in vivo findings. We use isolectin B4 staining to detect and, subsequently, quantify neovascularization and collagen 1a for staining of fibrosis.
- Ragauskas S, Kielczewski E, Vance J, Kaja S, Kalesnykas G. In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model. JoVE 2018; 131.
- Kaja S, Ragauskas S, Vähätupa M, Cerrada-Gimenez M, Mering S, Hakkarainen JJ, Kalesnykas G (2019) Standardization and validation of intravitreal and systemic administration of aflibercept in preclinical models of angiogenesis. Poster presentation at ARVO 2019.
- Dobi, E. T., Puliafito, C. A., & Destro, M. A new model of experimental choroidal neovascularization in the rat. Arch. Ophthalmol. Chic. Ill 1960. 107, 264-269 (1989).
- Grossniklaus HE, Kang SJ, Berglin L. Animal models of choroidal and retinal neovascularization. Prog Retin Eye Res (2010). 29:500-19. PubMed PMID: 20488255.