Performing Injection into Orbital Lobe of Superior Lacrimal Gland: A Technique for Inducing Dry Eye Disease in Rabbit Model

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Animal preparation

NOTE: The portions of the orbital lacrimal gland system are injected.

1. Sedate the rabbits with acepromazine 0.2 mg/kg subcutaneously.
2. Shear off the fur in the periorbital and scalp area and completely remove any residual fur using Nair. Leave the skin entirely smooth for better visualization of the anatomical hallmarks and US-guided injection of concanavalin A (Figure 1).
3. Induce moderate sedation as described above.

2. Injection of the orbital superior lacrimal gland (OSLG)

NOTE: OSLG follows in rapid succession.

1. Apply medial pressure to the globe causing the OSLG to protrude from the posterior incisure. Apply medial pressure to the globe (Figure 2, red arrow) with the protuberance of the OSLG from the posterior incisure. The protuberance serves as gross localization to find the posterior incisure.
2. Use curved forceps with tips closed to indent the area until the bony opening in the skull is felt. This will be slit-like with an anterior/posterior direction under the protuberance.
3. Apply modest pressure with forceps to leave an indentation in the skin, which will serve as the landmark for needle placement (Figure 3A).
4. Insert a needle (tuberculin syringe with a 27-gauge, 5/8-inch needle) perpendicular to the skin over the indentation mark (Figure 3B) ~ ¼ inch into the incisure, then redirect the needle posteriorly and externally towards the lateral canthus aiming for the midpoint between the injection site and bony orbital rim.
   NOTE: If the incisure is not precisely targeted with the needle, the skull blocks its advancement.
5. Once the hub of the needle is reached, slowly inject 1000 µg of Con A in a volume of 0.2 mL (Figure 3C).
6. Complete the injection of both the PSLG and OSLG within 2-3 min
7. Remove the animal from isoflurane sedation (if not yet done). Injection of the inferior lacrimal gland (ILG) can usually be completed without further sedation.