Western Blotting Assay for Egr-1 Immediate Early Gene in Brain Tissue of Zebrafish Applied to Neuroethological Study

Egr-1 (also known as zif268 , Krox-24 , NGFI-A, and ZENK) has been one of the most widely immediate early genes used to map specific neuronal populations activated during behavior in mammals and birds. However, the use of egr-1 mapping technique in nontraditional model systems depends on the evolutionary conservation of this gene. In the current chapter, we describe the western blotting technique for egr-1 in brain tissue of adult zebrafish (using 30 μg protein in the sample) applied to a behavioral study. As an example we introduce the chemical stimulation of alarm substance in the tank, and evaluated the activation of neurons in forebrain. However, this protocol can also be helpful to study neural activation in other behavioral context elicited by distinct stimuli. In addition, this method can be applied for other brain areas.