A Method to Induce Ocular Surface Inflammation in a Murine Model

Published: August 31, 2023

Abstract

Source: Singh, J., et al. Induction of Ocular Surface Inflammation and Collection of Involved Tissues. J. Vis. Exp. (2022)

This video demonstrates a method to induce ocular surface inflammation and meibomian gland dysfunction in a murine model. Stimulation of the mouse's immune system using an immunogen and later restimulation with the same immunogen promotes ocular surface inflammation, which causes plugging of meibomian gland secretion through neutrophil-produced aggNETs and leads to dry eye disease.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Induction of murine ocular surface inflammation Perform immunogen preparation for immunization. Prepare the immunogen freshly on the day of immunization, mixing ovalbumin (OVA, 50 mg/mL) and pertussis toxin (100 µg/mL) in saline and the adjuvant aluminum hydroxide (40 mg/mL) (see Table of M…

Representative Results

Figure 1: Excision of the facial tissue surrounding the orbital region. The incision trail is indicated in red. This allows for excising the ocular organs and investigating the influence and effects of various topically administered therapeutics on accessory ocular organs such as the conjunctiva and cornea.

Declarações

The authors have nothing to disclose.

Materials

1x PBS Gibco
Aluminium Hydroxide Imject alum Adjuvant 77161 40 mg/ mL
Final Concentration: in vivo: 1 mg/ 100 µL
C57Bl/6 mice, aged 7–9 weeks Charles River Laboratories
Curved forceps FST by Dumont SWITZERLAND 5/45 11251-35
Fine sharp scissor FST Stainless steel, Germany 15001-08
Laminar safety cabinet Herasafe
NaCl 0.9% (Saline) B.Braun
Ovalbumin (OVA) Endofit, Invivogen 9006-59-1 10 mg/200 µL in saline
Pertussis toxin ThermoFisher Scientific PHZ1174 50 µg/ 500 µL in saline
Final Concentration: in vivo: 100 µg/ 100 µL
Petridish Greiner bio-one 628160
Scalpel Feather disposable scalpel No. 21  Final Concentration: in vivo:  300 ng/ 100 µL
Stereomicroscope Zaiss Stemi508
Syringe (corneal/iris washing) BD Microlane 27 G x 3/4 – Nr.20 0,4 x 19 mm
Syringe (i.p immunization) BD Microlane 24 G1"-Nr 17, 055* 25 mm

Tags

Play Video

Citar este artigo
A Method to Induce Ocular Surface Inflammation in a Murine Model. J. Vis. Exp. (Pending Publication), e21604, doi: (2023).

View Video