Effects of acute morphine withdrawal on ultrasonic vocalizations in adult rats: unchanged 50-kHz call rate and altered subtype profile

Adult rat 22- and 50-kHz ultrasonic vocalizations (USVs) are commonly considered as indices of negative and positive affect, respectively. More specifically, we have proposed that positive affective states are revealed by a predominance of trill over flat 50-kHz call subtypes. However, the 50-kHz call subtypes emitted during aversive drug states remain largely uninvestigated. To determine whether acute morphine withdrawal affects 50-kHz call rates or alters the relative prevalence of trill and flat calls. In experiment 1, adult male rats were given saline or morphine (6 mg/kg SC), then acutely challenged 4 h later with saline or naloxone (1 mg/kg SC), and recorded 10-30 min post-injection. In experiments 2 and 3, rats received saline or morphine (6 mg/kg), followed 4 h later by acute saline or naloxone (0.1 mg/kg) challenge; USVs were subsequently recorded during 30-min place conditioning sessions. Naloxone (0.1 mg/kg) produced a strong conditioned place aversion only after acute morphine pretreatment, consistent with antagonist-precipitated morphine withdrawal. The morphine-naloxone combination decreased the relative prevalence of trills and promoted flat calls. Naloxone given alone (0.1 and 1 mg/kg) inhibited trill calls but did not significantly alter the prevalence of flat calls, whereas morphine given alone (4 h pre-session) was largely without effect. Fifty-kHz call rates were inhibited by naloxone given alone, but otherwise unaffected. Twenty-two-kHz calls were sparse. The 50-kHz call subtype shift seen during antagonist-precipitated morphine withdrawal was opposite in direction to that previously associated with rewards, and hence may reveal negative affect.