Steven M. Phelps

Animal display behaviors, such as advertisement songs, are flashy and attention grabbing by necessity. In order to balance the costs and benefits of such signals, individuals must be able to assess both their own energetic state and their social environment. In this study, we investigated the role of leptin, a hormonal signal of high energy balance, in regulating the vocal advertisement display of Alston’s singing mouse ( Scotinomys teguina ). We manipulated perception of energy balance using exogenous leptin, and social environment through acoustic playback, to ask how internal cues of energy availability are integrated with external social cues that promote singing. We found that both song playback and leptin injection promoted increased song effort. In the absence of song playback, leptin altered amplitude modulation in songs, but did not affect song rate. Additionally, we examined hormone and playback effects on non-vocal behaviors and found that leptin may shift physical activity away from cage exploration and toward wheel running. Finally, we found several positive associations between measures related to high song effort. These results demonstrate that male singing mice use both social context and energy balance to govern their investment in advertisement song and that leptin acts as a mediator of this process.

Identifying variation in vocal morphology and sound production mechanisms is essential to understanding vocal diversity. Rodents provide an ideal system for exploring this variation as they use multiple sound production mechanisms and have novel vocal structures whose morphology varies interspecifically. Here, we describe the laryngeal morphology and identify the sound production mechanism of Alston’s singing mouse ( Scotinomys teguina ), which produce stereotyped songs. We used micro-computed tomography to examine laryngeal morphology and manipulated excised larynges and surgically ablated a laryngeal muscle to determine sound production mechanism and frequency control. Laryngeal manipulations indicated that a whistle mechanism, likely an edge tone or shallow cavity, produces song. Singing mouse whistles are unique compared to other rodents because they rely on the inflation of an enlarged intralaryngeal air sac called the ventral pouch. Whistle frequency can be controlled by ventral pouch inflation, laryngeal airflow, and by cricothyroid muscle action. Cricothyroid ablation inhibited frequency modulation in vivo , suggesting that singing mice use this muscle during singing. Singing mouse laryngeal morphology and vocal mechanism are distinct from other Neotomids; instead of using vocal fold oscillations for loud, long-distance calls and whistles for close-range interactions, singing mice appear to use whistles for distant and close exchanges by inflating their intralaryngeal air sac. Air sacs have evolved repeatedly among vocalizing mammals and our results indicate a new role for these structures in generating sound. Together, our results expand on an emerging story of how biomechanic and morphological variation contributes to vocal diversity.

Animal courtship and aggressive displays are dramatic, often sexually dimorphic behaviors that require the coordinated modulation of diverse motivational and motor circuits. In Alston’s singing mice, a novel and elaborate advertisement vocalization is sexually dimorphic and steroid sensitive ( Scotinomys teguina ). Males sing more often than females, and on average male songs have more notes. Song is influenced by circulating androgens, but how such hormonal differences influence the diverse brain regions involved in vocal display is not understood. To characterize androgen-sensitive sites in the vocal motor pathway, we used two isoforms of pseudorabies virus (PRV) to double-label circuits ending in laryngeal and jaw muscles involved in vocalization, and co-labeled these neurons for androgen receptor (AR). Next we manipulated circulating androgens and observed the effects on AR distribution and male song. We find androgens drive coordinated changes in AR abundance across motor and motivational circuits, and both individual and group differences in song are associated with AR abundance. The results reveal how circulating androgens and the auto-regulation of androgen receptors can influence the diverse circuits necessary for elaborate advertisement displays. Significance statement: Courtship and aggressive displays are among those most elaborate and dramatic of sexually dimorphic behaviors. We show that in Alston’s singing mouse, an androgen-sensitive network defined by retrograde tracing shapes vocal display effort. Our results reveal how the intensity of singing mouse vocalizations is influenced by androgen actions in the vocal motor pathway.

Sexual bonds are central to the social lives of many species, including humans, and monogamous prairie voles have become the predominant model for investigating such attachments. We developed an automated whole-brain mapping pipeline to identify brain circuits underlying pair-bonding behavior. We identified bonding-related c-Fos induction in 68 brain regions clustered in seven major brain-wide neuronal circuits. These circuits include known regulators of bonding, such as the bed nucleus of the stria terminalis, paraventricular hypothalamus, ventral pallidum, and prefrontal cortex. They also include brain regions previously unknown to shape bonding, such as ventromedial hypothalamus, medial preoptic area, and the medial amygdala, but that play essential roles in bonding-relevant processes, such as sexual behavior, social reward, and territorial aggression. Contrary to some hypotheses, we found that circuits active during mating and bonding were largely sexually monomorphic. Moreover, c-Fos induction across regions was strikingly consistent between members of a pair, with activity best predicted by rates of ejaculation. A novel cluster of regions centered in the amygdala remained coordinated after bonds had formed, suggesting novel substrates for bond maintenance. Our tools and results provide an unprecedented resource for elucidating the networks that translate sexual experience into an enduring bond.

Sexual bonds are central to the social lives of many species, including humans, and monogamous prairie voles have become the predominant model for investigating such attachments. We developed an automated whole-brain mapping pipeline to identify brain circuits underlying pair-bonding behavior. We identified bonding-related c-Fos induction in 68 brain regions clustered in seven major brain-wide neuronal circuits. These circuits include known regulators of bonding, such as the bed nucleus of the stria terminalis, paraventricular hypothalamus, ventral pallidum, and prefrontal cortex. They also include brain regions previously unknown to shape bonding, such as ventromedial hypothalamus, medial preoptic area and the medial amygdala, but that play essential roles in bonding-relevant processes, such as sexual behavior, social reward and territorial aggression. Contrary to some hypotheses, we found that circuits active during mating and bonding were largely sexually monomorphic. Moreover, c-Fos induction across regions was strikingly consistent between members of a pair, with activity best predicted by rates of ejaculation. A novel cluster of regions centered in the amygdala remained coordinated after bonds had formed, suggesting novel substrates for bond maintenance. Our tools and results provide an unprecedented resource for elucidating the networks that translate sexual experience into an enduring bond.

Sexual bonds are central to the social lives of many species, including humans, and monogamous prairie voles have become the predominant model for investigating such attachments. We developed an automated whole-brain mapping pipeline to identify brain circuits underlying pair-bonding behavior. We identified bonding-related c-Fos induction in 68 brain regions clustered in seven major brain-wide neuronal circuits. These circuits include known regulators of bonding, such as the bed nucleus of the stria terminalis, paraventricular hypothalamus, ventral pallidum, and prefrontal cortex. They also include brain regions previously unknown to shape bonding, such as ventromedial hypothalamus, medial preoptic area and the medial amygdala, but that play essential roles in bonding-relevant processes, such as sexual behavior, social reward and territorial aggression. Contrary to some hypotheses, we found that circuits active during mating and bonding were largely sexually monomorphic. Moreover, c-Fos induction across regions was strikingly consistent between members of a pair, with activity best predicted by rates of ejaculation. A novel cluster of regions centered in the amygdala remained coordinated after bonds had formed, suggesting novel substrates for bond maintenance. Our tools and results provide an unprecedented resource for elucidating the networks that translate sexual experience into an enduring bond.

Although vocalizations are essential to mammalian sociosexual behaviours, little is known about female preferences for male vocal displays in non-model mammalian species. Here, we characterized female preferences for the advertisement songs of male Alston’s singing mice (Scotinomys teguina). We developed procedures for inducing oestrus, using vaginal perforation as a morphological indicator of oestrus. We then broadcasted ‘high-effort’ and ‘low-effort’ songs recorded from wild-caught males to virgin laboratory-reared females, using a two-choice experiment. Our results indicate that females spent more time investigating speakers playing high-effort songs; surprisingly, this phonotactic response is independent of oestrus status. In wild singing mice, acoustic characteristics of high-effort male songs positively correlate with body condition. Our data suggest that females could use acoustic cues to select mates in good condition, thus providing preliminary support for adaptive mate choice hypotheses. More generally, our results support the hypothesis that elaborate Scotinomys song may be shaped by female choice.

Background: Developing genomic resources for a diverse range of species is an important step towards understanding the mechanisms underlying complex traits.Specifically, organisms that exhibit unique, accessible phenotypes-of-interests, allow researchers to address questions that may be ill-suited to traditional model organisms. We sequenced the genome and transcriptome of Alston’s singing mouse ( Scotinomys teguina ), an emerging model for social cognition and vocal communication. In addition to producing advertisement songs used for mate attraction and male-male competition, these rodents are diurnal, live at high-altitudes, and are obligate insectivores, providing opportunities to explore diverse physiological, ecological, and evolutionary questions. Results: Using PromethION, Illumina, and PacBio sequencing, we produced an annotated genome and transcriptome, which were validated using gene expression and functional enrichment analyses. To assess the usefulness of our assemblies, we performed single nuclei sequencing on cells of the orofacial motor cortex, a brain region implicated in song coordination, identifying 12 cell types. Conclusions: These resources will provide the opportunity to identify the molecular basis of complex traits in singing mice as well as to contribute data that can be used for large-scale comparative analyses.

Vocalizations are often elaborate, rhythmically structured behaviors. Vocal motor patterns require close coordination of neural circuits governing the muscles of the larynx, jaw, and respiratory system. In the elaborate vocalization of Alston's singing mouse (Scotinomys teguina) each note of its rapid, frequency‐modulated trill is accompanied by equally rapid modulation of breath and gape. To elucidate the neural circuitry underlying this behavior, we introduced the polysynaptic retrograde neuronal tracer pseudorabies virus (PRV) into the cricothyroid and digastricus muscles, which control frequency modulation and jaw opening, respectively. Each virus singly labels ipsilateral motoneurons (nucleus ambiguus for cricothyroid, and motor trigeminal nucleus for digastricus). We find that the two isogenic viruses heavily and bilaterally colabel neurons in the gigantocellular reticular formation, a putative central pattern generator. The viruses also show strong colabeling in compartments of the midbrain including the ventrolateral periaqueductal gray and the parabrachial nucleus, two structures strongly implicated in vocalizations. In the forebrain, regions important to social cognition and energy balance both exhibit extensive colabeling. This includes the paraventricular and arcuate nuclei of the hypothalamus, the lateral hypothalamus, preoptic area, extended amygdala, central amygdala, and the bed nucleus of the stria terminalis. Finally, we find doubly labeled neurons in M1 motor cortex previously described as laryngeal, as well as in the prelimbic cortex, which indicate these cortical regions play a role in vocal production. The progress of both viruses is broadly consistent with vertebrate‐general patterns of vocal circuitry, as well as with circuit models derived from primate literature.

Social behavior varies across both individuals and species. Research to explain this variation falls under the purview of multiple disciplines, each with its own theoretical and empirical traditions. Integration of these disciplinary traditions is key to developing a holistic perspective. Here, we review research on the biology of social attachment, a phenomena in which individuals develop strong affective connections to one another. We provide a historical overview of research on social attachment from psychological, ethological and neurobiological perspectives. As a case study, we describe work on pair‐bonding in prairie voles, a socially monogamous rodent. This specific topic takes advantage of many biological perspectives and techniques to explain social bonds. Lastly, we conclude with an overview of multi‐dimensional conceptual frameworks that can be used to explain social phenomena, and we propose a new framework for research on individual variation in attachment behavior. These conceptual frameworks originate from philosophy, physics, ethology, cognitive science and neuroscience. The application and synthesis of such frameworks offers a rich opportunity to advance understanding of social behavior and its mechanisms.