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