Protocol for lens removal in embryonic fish and its application on the developmental effects of eye regression

Abstract

The lens plays a central role in the development of the optic cup. In fish, regression of the eye early in development affects the development of the craniofacial skeleton, the size of the olfactory pits, the optic nerve, and the tectum. Lens removal further affects olfaction, prey capture, and aggression. The similarity of the fish eye to other vertebrates is the basis for its use as an excellent animal model of human defects. Questions regarding the effects of eye regression are specifically well-suited to be addressed by using fish from the genus Astyanax. The species has two morphs; an eyeless cave morph and an eyed, surface morph. In the cavefish, a lens initially develops in embryos, but then degenerates by apoptosis. The cavefish retina is subsequently disorganized, degenerates, and retinal growth is arrested. The same effect is observed in surface fish when the lens is removed or exchanged for a cavefish lens. While studies can greatly benefit from a control group of surface fish with regressed eyes brought through lensectomies, few studies include them because of technical difficulties and the low survivorship of embryos that undergo this procedure. Here we describe a technique with significant modification for improvement for conducting lensectomy in one-day-old Astyanax and other fish, including zebrafish. Yields of up to 30 live embryos were obtained using this technique from a single spawn, thus enabling studies that require large sample sizes.