The composition of the vehicle saline was as follows (all chemicals were obtained from Sigma): 111 mM NaCl, 2 mM KCl, 2 mM CaCl2, 1 mM MgSO4, 1 mM NaHCO3, and 0.5 Mm NaH2PO4. The pH of the saline solution was 6.8. Glutamate (Sigma) and the 5-HT2 receptor antagonist KET (Sigma) were dissolved in saline before application, as done previously (27). Drug application electrodes were made using two-barrel KG-33 glass micropipettes [outer diameter (OD), 1.5 mm; internal diameter (ID), 0.86 mm; A-M Systems] and pulled by a vertical micropipette puller (Stoelting) to a fine tip that was subsequently broken to attain a tip diameter of approximately 5 μm for each barrel. The two barrels were used for separate application of either KET (100 mM) or NBOH (100 μM), as well as glutamate (1 mM). During ELL recordings for which we injected KET, we first used excitatory responses to glutamate application to confirm that we were within proximity of the pyramidal neuron we were recording from, as done previously (24, 38). KET (n = 8 PCells; average recording time, 121.8 ± 1.1 min) was then ejected to the neuron to ensure a local effect. For behavioral recordings, injections of KET (n = 8 fish; average recording time, 240 ± 22.7 min) were performed bilaterally in ELL, as done previously (12, 13, 24). All pharmacological injections were performed using a duration of 130 ms at ~20 psi using a Picospritzer (General Valve) as done previously (23). For pharmacological experiments using KET, injections were done every 10 min during the adaptation period. We note that injecting saline alone as a control did not alter behavioral or neuronal activity as shown in previous studies (24, 39).

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