Abstract
Rats emit ultrasonic vocalizations (USVs) in situations with emotional valence, and USVs have also been proposed as a marker for memories conditioned to those situations. This study investigated whether USV emissions can predict and/or be associated with the behavior of rats in tests that evaluate unconditioned memory. To this end, rats were subjected to “tickling”, a procedure of heterospecific play that has emotional valence and elicits the emission of USVs, and afterwards evaluated in the novel object recognition test (NOR) and in the single trial continuous spontaneous alternation behavior (SAB) test in a Y maze. The number of 22-kHz USVs (aversive) and 50-kHz USVs (appetitive) emitted in response to tickling and during NOR and SAB tests were scored, and the correlations among them and with rats’ behavior evaluated. Rats emitted 50-kHz USVs, but not 22-kHz USVs, during the NOR and SAB tests, and such calling behavior was not linked with the behavioral readouts indicative of memory function in either test. However, rats that prevalently emitted 22-kHz USVs in response to tickling displayed an impaired NOR performance. These findings suggest that measuring the emission of USVs could be of interest in studies of unconditioned memory, at least with regard to 22-kHz USVs.
Highlights
Rats emit ultrasonic vocalizations (USVs) in response to and/or anticipation of various stimuli that possess emotional valence [1,2,3]
USV data were analyzed by an experimenter blind to th3e.2c.oEnmdiitsisoinons ooff Urelctroarsdoinnigc .VAoclladlizaatatioonbstaininRedespinonthseetporTeisceknlitnsgtudy were tested for normality witThicthkleinKgoslmignoigfiocraonvt–lySmstiimrnuolvatteedstthanedemanisasliyozneodfaUccSoVrsdiinngralytsw
The Kruskal–Wallis test showed the presence of significant modifications in the cumulative number of categorized 50-kHz USVs emitted over the 5 days of tickling (K = 71.43, p < 0.01), and Dunn’s test for multiple comparisons revealed that a significant increase in the emission of FM (p < 0.01) and flat (p < 0.01) calls occurred during tickling sessions, compared with baseline sessions when the hand of the experimenter was passively left inside the cage (Figure 3C)
Summary
Rats emit ultrasonic vocalizations (USVs) in response to and/or anticipation of various stimuli that possess emotional valence [1,2,3]. Situations that possess positive emotional valence for rats (i.e., non-aggressive interactions with conspecifics or familiar humans and administration of drugs with rewarding properties) elicit the emission of the so-called 50-kHz USVs, which have a short duration (generally 30–40 ms), sound frequency between 35–80 kHz (which can be even higher), and may possess marked frequency modulation [4]. Rats may emit either anticipatory 22-kHz USVs when re-exposed to an environment where they received aversive electric footshocks [7,8], or anticipatory 50-kHz USVs in response to environmental cues previously associated with pharmacological or non-pharmacological rewards [9,10,11,12,13,14,15,16,17] Taken together, these findings have suggested that calling behavior may be a marker of conditioned memories for stimuli/situations that possess emotional valence. Measuring USVs could potentially complement the classical behavioral markers that are evaluated in rat studies of memory (i.e., preference for items, patterns of arm exploration in mazes), increasing the quantity of information that can be collected in those studies
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