*Article* **Nucleus Accumbens Chemogenetic Inhibition Suppresses Amphetamine-Induced Ultrasonic Vocalizations in Male and Female Rats**

**Kate A. Lawson † , Abigail Y. Flores † , Rachael E. Hokenson † , Christina M. Ruiz and Stephen V. Mahler \***

> Department of Neurobiology & Behavior, University of California, Irvine. 1203 McGaugh Hall, Irvine, CA 92697, USA; lawsonk1@uci.edu (K.A.L.); abigaiyf@uci.edu (A.Y.F.); rhokenso@uci.edu (R.E.H.); cruiz1@uci.edu (C.M.R.)

**\*** Correspondence: mahlers@uci.edu

† Denotes equal contributions.


**Citation:** Lawson, K.A.; Flores, A.Y.; Hokenson, R.E.; Ruiz, C.M.; Mahler, S.V. Nucleus Accumbens Chemogenetic Inhibition Suppresses Amphetamine-Induced Ultrasonic Vocalizations in Male and Female Rats. *Brain Sci.* **2021**, *11*, 1255. https://doi.org/10.3390/ brainsci11101255

Academic Editors: Stefan M. Brudzynski and Jeffrey Burgdorf

Received: 28 July 2021 Accepted: 17 September 2021 Published: 22 September 2021

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**Abstract:** Adult rats emit ultrasonic vocalizations (USVs) related to their affective states, potentially providing information about their subjective experiences during behavioral neuroscience experiments. If so, USVs might provide an important link between invasive animal preclinical studies and human studies in which subjective states can be readily queried. Here, we induced USVs in male and female Long Evans rats using acute amphetamine (2 mg/kg), and asked how reversibly inhibiting nucleus accumbens neurons using designer receptors exclusively activated by designer drugs (DREADDs) impacts USV production. We analyzed USV characteristics using "Deepsqueak" software, and manually categorized detected calls into four previously defined subtypes. We found that systemic administration of the DREADD agonist clozapine-n-oxide, relative to vehicle in the same rats, suppressed the number of frequency-modulated and trill-containing USVs without impacting high frequency, unmodulated (flat) USVs, nor the small number of low-frequency USVs observed. Using chemogenetics, these results thus confirm that nucleus accumbens neurons are essential for production of amphetamine-induced frequency-modulated USVs. They also support the premise of further investigating the characteristics and subcategories of these calls as a window into the subjective effects of neural manipulations, with potential future clinical applications.

**Keywords:** 50 kHz vocalizations; 22 kHz vocalizations; amphetamine; nucleus accumbens; chemogenetics; clozapine-n-oxide; UMAP; males; females
