Developmental neuroscienceA choice behavior for morphine reveals experience-dependent drug preference and underlying neural substrates in developing larval zebrafish
Section snippets
Rearing
Adult zebrafish were maintained following standard procedures (Westerfield, 1995). AB is a laboratory-bred strain from the University of Oregon. EK is a commercially available strain from Ekkwill, Gibsonton, FL, USA. Larvae of AB wild type or AB/EK hybrid strain (backcrossed to AB for more than five generations) were raised by groups of 50 until the age of 2 weeks at 28 °C in a rearing tank (25×16.5×6.5 cm), with 1 l of water containing 3 g of artificial Instant Ocean salts (Aquatic Ecosystem,
Larval zebrafish display a preference for morphine in a choice chamber paradigm
We devised a choice chamber as described in Experimental Procedures (Fig. 1A). Ten minutes after starting the flows, morphine was detected only in half of the chamber nearest the source of delivery (Table 1). Therefore, by monitoring their location in the tank, we could directly assess whether larval zebrafish display morphine preference: if more larvae spent their time in the morphine-containing side than in the water-containing side, it would serve as an indication that larvae have a
Discussion
Toward the goal of elucidating the molecular and cellular basis of reward-associated choice behavior, we have established a choice chamber paradigm in larval zebrafish: 2-week-old larval zebrafish exhibit an experience-dependent preference for the addictive drug morphine. Such behavior is not due to a peripheral effect of morphine, but is dependent on central opioid and monoamine neurotransmitter systems.
The choice behavior is measured by a preference of larval zebrafish to self-immerse in the
Acknowledgments
We thank Drs. Antonello Bonci, Howard Fields, Ulrike Heberlein, Patricia Janak, Larry Tecott, and Fred Wolf for helpful comments or advice on behavioral analyses, Billy Lau for statistical help, and other members of the Guo laboratory for discussions. We also thank Drs. Gil Levkowitz and Arnon Rosenthal for sharing the too few gene sequence information prior to publication, and the National Institute of Drug Abuse for providing us with morphine sulfate. This work was supported by grants from
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