Development of an Aversive Conditioning Task Using a "Tapping" Method for Juvenile Danio rerio

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In recent years, Danio rerio (zebrafish) have increasingly been used as a laboratory organism to model and investigate CNS diseases and vertebrate behaviour. Several studies have analyzed learning and memory in adult zebrafish. However, juvenile zebrafish have only rarely been utilized for this purpose due to the paucity of learning paradigms. One of the first steps in developing a novel learning task is to identify appropriate reinforcers, unconditioned stimuli that motivate the subject. We have developed a simple method with which we can generate vibration in a consistent manner. The vibration is expected to generate low frequency waves in the water detectable by the lateral line of the fish. We describe this method, and show that the vibration induces measurable fear responses in juvenile (10-12 days post-fertilization old) zebrafish. The task utilizes “tapping” achieved by a rubberized metal ball attached to a nylon string released from a frame at a set distance from the side glass wall of the test tank. We employed this tapping once every 2 minutes for a total of five occasions per test session. We compared the behavior of our experimental fish one-second before and one second immediately after each tap, and found significant differences between pre- and post-tap intervals for total distance travelled and relative turn angle. These differences remained consistent throughout all five taps of the session. Subsequently, we developed a learning task utilizing the tapping method. In this task, juvenile zebrafish were required to distinguish the blue and the green side of a circular arena with one of these sides negatively reinforced by tapping, an aversive visual discrimination task. We describe the detailed methodology of this learning task and will report learning and memory parameters once the study has been completed.

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