The use of supervised machine learning to approximate poses in video recordings allows for rapid and efficient analysis of complex behavioral profiles. Currently, there are limited protocols for automated analysis of operant self-administration behavior. We provide a methodology to (1) obtain videos of training sessions via Raspberry Pi microcomputers or GoPro cameras, (2) obtain pose estimation data using the supervised machine learning software packages DeepLabCut (DLC) and Simple Behavioral Analysis (SimBA) with a local high-performance computer cluster, (3) compare standard Med-PC lever response versus quadrant time data generated from pose estimation regions of interest, and (4) generate predictive behavioral classifiers. Overall, we demonstrate proof of concept to use pose estimation outputs from DLC to both generate quadrant time results and obtain behavioral classifiers from SimBA during operant training phases.

Food intake is controlled by multiple converging signals: hormonal signals that provide information about energy homeostasis and hedonic and motivational aspects of food and food cues that can drive nonhomeostatic or “hedonic” feeding. The ventral pallidum (VP) is a brain region implicated in the hedonic and motivational impact of food and food cues, as well as consumption of rewards. Disinhibition of VP neurons has been shown to generate intense hyperphagia, or overconsumption. While VP GABA neurons have been implicated in cue-elicited reward-seeking and motivation, the role of these neurons in the hyperphagia resulting from VP activation remains unclear. Here, we used designer receptors exclusively activated by designer drugs to activate VP GABA neurons in nonrestricted male and female rats during chow and sucrose consumption. We found that activation of VP GABA neurons increases consumption of chow and sucrose in male rats, but not female rats. Together, these findings suggest that activation of VP GABA...

Chronic pain is a debilitative disease affecting one in five adults globally and is a major risk factor for anxiety ( [Goldberg and McGee, 2011][1]; [Lurie, 2018][2]). Given the current dearth of available treatments for both individuals living with chronic pain and mental illnesses, there is a critical need for research into the molecular mechanisms involved in order to discover novel treatment targets. Cellular homeostasis is crucial for normal bodily functions, and investigations of this process may provide better understanding of the mechanisms driving the development of chronic pain. Using the spared nerve injury (SNI) model of neuropathic pain, we found contrasting roles for BECLIN-1 in the development of pain hypersensitivity and anxiety-like behaviors in a sex-dependent manner. Remarkably, we found that male SNI mice with impaired BECLIN-1 function demonstrated heightened mechanical and thermal hypersensitivity compared with male wild-type SNI mice, while female SNI mice with impaired BECLIN-1 func...

The behavioral interactions between adults and newborns are decisive for the fitness and the survival of offspring across the animal kingdom. In laboratory mice, while virgin females display caregiving behaviors, virgin males are rather neglectful or aggressive toward pups. Despite the importance of these behavioral variations, the underlying neural mechanisms remain poorly understood. Brain regions encoding these behaviors may exhibit sex-dependent functional differences at the baseline. Additionally, these structures might undergo sex-specific plasticity after adults interact with the offspring. Emerging evidence suggests sex-based differences in input connectivity, genetics, and receptor expression of the epithalamic lateral habenula (LHb). Moreover, LHb neuronal activity is instrumental for adult–newborn interactions. However, whether LHb neuronal function varies between sexes and/or undergoes adaptations following interactions with pups has not been fully investigated. In this study, we used in vivo a...

Functional imaging studies indicate that both the assessment of a person as untrustworthy and the assumption that a person has a sexually transmitted infection are associated with activation in regions of the salience network. However, studies are missing that combine these aspects and investigate the perceived trustworthiness of individuals previously assessed with high or low probability of a sexually transmitted infection. During fMRI measurements, 25 participants viewed photographs of people preclassified as having high or low HIV probability and judged their trustworthiness. In a postrating, stimuli were rated for trustworthiness, attractiveness, and HIV probability. Persons preclassified as HIV− in contrast to those preclassified as HIV+ were rated more trustworthy and with lower HIV probability. Activation in medial orbitofrontal cortex was higher for those rated and preclassified as HIV− than HIV+. Based on the individual ratings, but not the preclassification, there was significantly higher activa...

Breathing is a complex neuromuscular process vital to sustain life. In preclinical animal models, the study of respiratory motor control is primarily accomplished through neurophysiologic recordings and functional measurements of respiratory output. Neurophysiologic recordings that target neural or muscular output via direct nerve recordings or respiratory muscle electromyography (EMG) are commonly collected during anesthetized conditions. While offering tight control of experimental preparations, the use of anesthesia results in respiratory depression, may impact cardiovascular control, eliminates the potential to record volitional nonventilatory behaviors, and can limit translation. Since the diaphragm is a unique muscle which is rhythmically active and difficult to access, placing diaphragm EMGs to collect chronic recordings in awake animals is technically challenging. Here, we describe methods for fabricating and implanting indwelling diaphragm EMG electrodes to enable recordings from awake rodents for...

The adrenal medulla is packed with chromaffin cells, modified postganglionic sympathetic neurons that secrete the catecholamines, epinephrine and norepinephrine, during the fight-or-flight response. Sometimes overlooked is a population of immune cells that also resides within the gland but whose distribution and function are not clear. Here I examine the location of CD45+ hematopoietic cells in the mouse adrenal medulla and show the majority are F4/80+/Lyz2+ macrophages. These cells are present from early postnatal development and widely distributed. Anatomically they are associated with chromaffin cells, found aligned alongside synapsin-IR neuronal varicosities and juxtaposed to CD31-IR blood vessels. Using Lyz2cre-GCaMP6f mice to quantify calcium signaling in macrophages revealed these cells respond directly and indirectly to a wide variety of neuromodulators, including pre- and postganglionic transmitters and systemic hormones. Purinergic agonists, histamine, acetylcholine, and bradykinin rapidly and re...

Inner speech refers to the silent production of language in one’s mind. As a purely mental action without obvious physical manifestations, inner speech has been notoriously difficult to quantify. To address this issue, the present study repurposed the phenomenon of speaking-induced suppression, wherein overt speech has been consistently shown to elicit reduced auditory evoked potentials compared with externally generated speech, as well as changes in oscillatory activity in gamma and theta frequency bands. Given the functional similarities between inner and overt speech, we used an established experimental protocol to investigate whether similar metrics could be used to distinguish the content of inner speech. Healthy participants ( n = 129) produced an inner syllable at a precisely specified time. An audible syllable was concurrently presented which either matched or mismatched the content of the inner syllable. The results revealed that Match and Mismatch conditions could be differentiated on the basis o...

The amygdala is believed to make invaluable contributions to visual emotion processing. Yet how this subcortical body contributes to emotion perception across time is contended. Here, we measured differences in the perceptual processing of emotional stimuli after unilateral temporal lobe and amygdala resection (TLR) in humans, using EEG. Through mass univariate analysis of brain activity, we compared responses to fearful and neutral faces (left TLR N  = 8, right TLR N  = 8, control N  = 8), and fearful and neutral bodies (left TLR N  = 9, right TLR N  = 9, control N  = 9). We found that TLR impaired the early-stage perceptual processing of emotional stimuli seen in the control group. Indeed, in controls a heightened responses to fearful faces was found in the 140–170 ms time window, over temporoparietal electrodes. This effect was also present in the left TLR group but disappeared in the right TLR group. For emotional bodies, brain activity was differentially sensitive to fearful stimuli at 90–120 ms in th...

For comprehensive anatomical analysis of a mouse brain, accurate and efficient registration of the experimental brain samples to a reference atlas is necessary. Here, we introduce Bell Jar, a semiautomated solution that can align and annotate tissue sections with anatomical structures from a reference atlas as well as detect fluorescent signals with cellular resolution (e.g., cell bodies or nuclei). Bell Jar utilizes Mattes mutual information-directed B-spline transformations to achieve precise alignments, even with damaged sample tissues. While user input remains a requirement for fine-tuning section matches, the platform streamlines the process, aiding rapid analyses in high-throughput neuroanatomy studies. As a standalone desktop application with a user-friendly interface, Bell Jar’s performance, which surpasses traditional manual and existing automated methods, can improve the reproducibility and throughput of histological analyses.