Intravitreal Injection of Adeno-Associated Virus to Assess Retinal Barrier Disruption in Mice

Begin with an anesthetized mouse model with a compromised inner limiting membrane, a barrier that separates the retina from the vitreous cavity.

Apply dilating agents to enlarge the pupil for better visualization. Then, apply a lubricant to prevent dryness.

Load an ultrafine needle–fitted syringe with an adeno-associated viral vector carrying a green fluorescent protein gene.

Insert the needle near the limbus of the eye into the vitreous cavity and inject the viral vector without touching internal eye structures.

Apply an antibacterial and an anti-inflammatory agent to the eye to prevent infection and inflammation.

Due to the increased permeability of the compromised inner limiting membrane, viral vectors penetrate into deeper retinal layers and infect retinal cells.

Infected retinal cells express the viral genome and produce GFP.

Under fluorescence imaging, green fluorescence from retinal cells confirms viral penetration through the compromised inner limiting membrane and infection of retinal cells.

Begin by anesthetizing C-57 black-six J mice. Check for the loss of three reflexes: the righting reflex, the withdrawal reflex, and tail pinch response.

Loss of these reflexes confirms anesthet