Modeling Retinal Degeneration and Regeneration in Zebrafish Using a Focal Laser Injury

Abstract

Source: Conedera, F. M., et al. Müller Glia Cell Activation in a Laser-induced Retinal Degeneration and Regeneration Model in Zebrafish. J. Vis. Exp. (2017).

This video demonstrates a procedure to induce retinal degeneration in adult zebrafish using laser injury. The laser targets photoreceptor cells in the retina, triggering cell death and a degenerative process. This damage activates Müller glia (MG) cells, leading to their dedifferentiation into stem cell progenitors that aid in retinal regeneration.

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. Maintain TgBAC (gfap:gfap-GFP) zebrafish 167 (AB) strain aged between 6 - 9 months under standard conditions in water with a temperature of 26.5 °C and a 14/10 h light/dark cycle.

2. Reversible Systemic Anesthesia

1. Prepare the stock solution of ethyl 3-aminobenzoate methanesulfonate salt (tricaine) by dissolving 400 mg of tricaine powder in 97.9 mL of tank water and 2.1 mL of 1 M of Tris buffered saline (TB....

Results

Figure 1: Setup for the induction of the laser injury of zebrafish retina and following OCT analysis. (A) Arrangement of the laser system before treatment. (A.1) Zebrafish placed on silicone pin holder with fundus contact len.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Hydroxypropylmethylcellulose 2%	OmniVision, Neuhausen, Switzerland	n.a.	Methocel 2%
Ethyl 3-aminobenzoate methanesulfonate	Sigma-Aldrich, Buchs, Switzerland	A5040	Tricaine, MS-222
Visulas 532s	Carl Zeiss Meditec AG, Oberkochen, Germany	n.a.	532 nm laser
Silicone pin holder	Huco Vision AG Switzerland	n.a.	Cut by hand from silicone pin mat of the sterilization tray accordingly.
TgBAC (gfap:gfap-GFP) zf167 (AB) strain	KIT, Karlsruhe, Germany	15204	http://zfin.org/ZDB-ALT-100308-3
Tris buffered saline (TBS)	Sigma-Aldrich, Buchs, Switzerland	P5912
Ocular fundus laser lens	Ocular Instruments, Bellevue, WA, USA	OFA2-0