A Method to Induce Ocular Surface Inflammation in a Murine Model

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

1. Perform immunogen preparation for immunization.
   1. 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 Materials) in a 1:1 proportion.
      CAUTION: Perform the opening, reconstitution, and immunogen preparation of pertussis toxin....

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 conj.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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