In Situ Hybridization in Zebrafish Larvae and Juveniles during Mesonephros Development

This protocol is significant because it allows the determination of gene expression in older zebrafish larvae and juveniles during metamorphosis. The advantage of this technique is that several steps have been optimized for probe penetration and visualization of the kidney. To begin, set up adult zebrafish to mate by adding one male and one female fish in a mating tank in the late afternoon after their last meal.

The next day, collect the embryos in Petri plates containing E3 medium. Five days post-fertilization, place a 400-micrometer screen in a 2.8-liter tank and fill it with 2 centimeters of system water. Then add the larva from one Petri plate.