Simultaneous video analysis of the kinematics of opercular movement and electromyographic activity during agonistic display in Siamese fighting fish

Neuroethology seeks to uncover the neural mechanisms underlying natural behaviour. One of the major challenges in this field is the need to correlate directly neural activity and behavioural output. In most cases, recording of neural activity in freely moving animals is extremely difficult. However, electromyographic recording can often be used in lieu of neural recording to gain an understanding of the motor output program underlying a well-defined behaviour. Electromyographic recording is less invasive than most other recording methods, and does not impede the performance of most natural tasks. Using the opercular display of the Siamese fighting fish as a model, we developed a protocol for correlating directly electromyographic activity and kinematics of opercular movement: electromyographic activity was recorded in the audio channel of a video cassette recorder while video taping the display behaviour. By combining computer-assisted, quantitative video analysis and spike analysis, the kinematics of opercular movement are linked to the motor output program. Since the muscle that mediates opercular abduction in this fish, the dilator operculi, is a relatively small muscle with several subdivisions, we also describe methods for recording from small muscles and marking the precise recording site with electrolytic corrosion. The protocol described here is applicable to studies of a variety of natural behaviour that can be performed in a relatively confined space. It is also useful for analyzing complex or rapidly changing behaviour in which a precise correlation between kinematics and electromyography is required.