A Hypertonic Saline Injection Technique for Inducing Glaucoma in Rats

Abstract

Source: Gossman, C. A. et al. Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation. J. Vis. Exp. (2016)

This video demonstrates glaucoma induction in an anesthetized rat by injecting hypertonic saline into the episcleral vein. The saline enters the trabecular meshwork via Schlemm's canal, causing tissue scarring, which blocks aqueous humor outflow and raises intraocular pressure. The elevated pressure damages retinal ganglion cells, leading to glaucoma.

Protocol

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

1. Animals

1. Use male and female rats 3 months of age in this study.
2. Keep animals in a 12 hr light/dark cycle with free access to food and water.

2. Preparation of KAX (Ketamine + Xylazine + Acepromazine) Cocktail for Animal Anesthesia

1. Dissolve 50 mg of xylazine (20 mg/ml) in 5 ml ketamine (100 mg/ml) with 1 ml acepromazine (10 mg/ml) and 3 ml of distilled water. Mix thoroughly.
2. Sterilize ....

Results

Figure 1. Microneedle Components. (A) Syringe with polyethylene tubing used for saline injection. (B) Backfill syringe used to backfill the borosilicate microneedle. (C) Narishige electrode puller used to make borosilicate.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Xylazine hydrochloride, Minimum 99%	Sigma, Life Science	X1251-1G
Ketamine hydrochloride injection, USP, 100mg/ml	Putney, Inc	NDC 26637-411-01	10 ml bottle
Acepromazine Maleate, 10mg/ml	Phoenix Pharmaceutical, Inc	NDC 57319-447-04, 670008L-03-0408	50 ml bottle
Serum bottle, 10 ml	VWR	16171319	Borosilicate glass
1 ml insulin syringe	VWR	BD329410	28 G needle
Sodium chloride	Sigma	S7653	2 M Solution
Microelectrode Puller	Narishige Group	PP-830
Heavy Polished Standard and Thin Walled Borosilicate Tubing	Sutter Instruments	B150-86-10HP	without filament, 0.86 mm
Microfil syringe needle for filling micropipettes	World Precision Instruments, Inc	MF28G
18 gauge Luer-Lock needle	Fisher Scientific	1130421	Syringe needle
Flexible Polyethylene Tubing	Fisher Scientific	22046941	0.034 inch diameter, approximately 10 inches
Proparacaine Hydrochloride Opthalmic Solution, USP, 0.5%	Akorn, Inc	NDC 17478-263-12	15 ml sterile bottle
Microscope	Leica	StereoZoom 4
Hemostat Clamp	Fine Science Tools	1310912	curved edge
Triple Antibiotic Ointment	Fisher Scientific	NC0664481
Scalpel handle	Fine Science Tools	10004-13
Scalpel blade #11	Fine Science Tools	10011-00
60 mm x 15 mm Disposable Petri Dish	VWR	351007
Phosphate Buffered Saline 10x Concentrate	Sigma, Life Science	P7059-1L	1x dilution
Spring Scissors	Fine Science Tools	15009-08
Forceps (2), Dumont # 5	Fine Science Tools	11251-30
Sylgard 184	VWR	102092-312