Protocol for implementing medial forebrain bundle stimulation as a reward for perceptual tasks in mice.

Training mice to perform perceptual tasks is a vital part of integrative neuroscience. Replacing classical rewards like water with medial forebrain bundle (MFB) stimulation allows experimenters to avoid deprivation and obtain higher trial numbers per session. Here, we provide a protocol for implementing MFB-based reward in mice. We describe steps for MFB electrode implantation, efficacy testing, and stimulation calibration. After these steps, MFB reward can be used to facilitate sensory discrimination task training and enable nuanced characterization of psychophysical abilities. For complete details on the use and execution of this protocol, please refer to Verdier et al. (2022).1.

Use of Digital Stethoscope to Measure Heart Rate in Birds: Comparison of Different Counting Methods Using Phonocardiograms.

The high cardiac contractility of birds poses a challenge to traditional cardiac auscultation, particularly for the accurate determination of heart rate (HR). The objectives of this study were to 1) evaluate the feasibility of using phonocardiograms of adequate length and quality to assess HR in different avian species with a commercially available digital stethoscope, 2) compare 5 counting methods, including 2 direct reading methods (manual counting and using a semiautomatic computerized algorithm as a reference method) and 3 listening methods (progressive mental counting, counting by 10s, and counting with a smartphone application tap counter), and 3) obtain the HR in selected birds and identify a correlation between body weight and HR in different avian species. An inverse correlation on a logarithmic scale was identified between the mean body weight and HR in 60 different bird species (n = 211; R = -0.72, P < 0.0001). Manual reading of phonocardiograms was the most reliable method and had the highest agreement with the reference method; this was followed by the counting by 10s method, the tapping method, and the progressive mental counting method, which was the least reliable. The agreement levels for the different methods were comparable for HRs <200 beats per minute (bpm) in birds weighing >1 kg. For HRs >500 bpm in birds weighing <150 g, only the reading method maintained a good agreement level. A digital stethoscope can be a useful tool for accurately determining the HR in birds, including very small species with high HRs.

Effect of Hooding on Physiological Parameters During Manual Restraint in Rhode Island Red Hybrid Hens (Gallus gallus domesticus).

Manual handling of chickens is required for many veterinary, research, and breeding procedures. This study aimed to assess the changes in physiological parameters over time during manual restraint of chickens, as well as the effect of hooding on these parameters. Heart rate, heart rate variability, respiratory rate, and body temperature were measured every 3 minutes for 15 minutes during manual restraint in 13 adult laying hens (Gallus gallus domesticus). Heart rate variability was significantly higher in hooded hens than in nonhooded hens (P= 0.003) but was not significant over time. Hooded hens were also found to have significantly lower heart rate (P = 0.043) and respiratory rate (P = 0.042) compared to nonhooded hens. Heart rate and respiratory rate significantly decreased over time, independent of the use of the hood (P = 0.008; P = 0.01, respectively). Temperature was found to increase significantly (P = 0.001) over time for both groups. Overall, hooding increased heart rate variability, a factor associated with a lower stress level, and decreased heart rate and respiratory rate. In conclusion, these data suggest that the use of the hood reduces stress levels in birds during manual restraint. Therefore, the use of the hood is encouraged for short (less than 15 minutes) painless procedures, such as physical examination or radiographic acquisition.

Enhanced perceptual task performance without deprivation in mice using medial forebrain bundle stimulation.

Perceptual decision-making tasks are essential to many fields of neuroscience. Current protocols generally reward deprived animals with water. However, balancing animals' deprivation level with their well-being is challenging, and trial number is limited by satiation. Here, we present electrical stimulation of the medial forebrain bundle (MFB) as an alternative that avoids deprivation while yielding stable motivation for thousands of trials. Using licking or lever press as a report, MFB animals learnt auditory discrimination tasks at similar speed to water-deprived mice. Moreover, they more reliably reached higher accuracy in harder tasks, performing up to 4,500 trials per session without loss of motivation. MFB stimulation did not impact the underlying sensory behavior since psychometric parameters and response times are preserved. MFB mice lacked signs of metabolic or behavioral stress compared with water-deprived mice. Overall, MFB stimulation is a highly promising tool for task learning because it enhances task performance while avoiding deprivation.

Breathing-driven prefrontal oscillations regulate maintenance of conditioned-fear evoked freezing independently of initiation.

Brain-body interactions are thought to be essential in emotions but their physiological basis remains poorly understood. In mice, regular 4 Hz breathing appears during freezing after cue-fear conditioning. Here we show that the olfactory bulb (OB) transmits this rhythm to the dorsomedial prefrontal cortex (dmPFC) where it organizes neural activity. Reduction of the respiratory-related 4 Hz oscillation, via bulbectomy or optogenetic perturbation of the OB, reduces freezing. Behavioural modelling shows that this is due to a specific reduction in freezing maintenance without impacting its initiation, thus dissociating these two phenomena. dmPFC LFP and firing patterns support the region's specific function in freezing maintenance. In particular, population analysis reveals that network activity tracks 4 Hz power dynamics during freezing and reaches a stable state at 4 Hz peak that lasts until freezing termination. These results provide a potential mechanism and a functional role for bodily feedback in emotions and therefore shed light on the historical James-Cannon debate.

The Theta Rhythm Mixes and Matches Gamma Oscillations Cycle by Cycle.

In the hippocampus, gamma power modulation by the theta rhythm is interpreted as the signature of temporally coordinated inputs that reflect ongoing processing. In this issue of Neuron, Lopes-Dos-Santos et al. (2018) develop a new methodology demonstrating that theta cycles can be viewed as individual computational units characterized by typical gamma profiles.

Harnessing olfactory bulb oscillations to perform fully brain-based sleep-scoring and real-time monitoring of anaesthesia depth.

Real-time tracking of vigilance states related to both sleep or anaesthesia has been a goal for over a century. However, sleep scoring cannot currently be performed with brain signals alone, despite the deep neuromodulatory transformations that accompany sleep state changes. Therefore, at heart, the operational distinction between sleep and wake is that of immobility and movement, despite numerous situations in which this one-to-one mapping fails. Here we demonstrate, using local field potential (LFP) recordings in freely moving mice, that gamma (50-70 Hz) power in the olfactory bulb (OB) allows for clear classification of sleep and wake, thus providing a brain-based criterion to distinguish these two vigilance states without relying on motor activity. Coupled with hippocampal theta activity, it allows the elaboration of a sleep scoring algorithm that relies on brain activity alone. This method reaches over 90% homology with classical methods based on muscular activity (electromyography [EMG]) and video tracking. Moreover, contrary to EMG, OB gamma power allows correct discrimination between sleep and immobility in ambiguous situations such as fear-related freezing. We use the instantaneous power of hippocampal theta oscillation and OB gamma oscillation to construct a 2D phase space that is highly robust throughout time, across individual mice and mouse strains, and under classical drug treatment. Dynamic analysis of trajectories within this space yields a novel characterisation of sleep/wake transitions: whereas waking up is a fast and direct transition that can be modelled by a ballistic trajectory, falling asleep is best described as a stochastic and gradual state change. Finally, we demonstrate that OB oscillations also allow us to track other vigilance states. Non-REM (NREM) and rapid eye movement (REM) sleep can be distinguished with high accuracy based on beta (10-15 Hz) power. More importantly, we show that depth of anaesthesia can be tracked in real time using OB gamma power. Indeed, the gamma power predicts and anticipates the motor response to stimulation both in the steady state under constant anaesthetic and dynamically during the recovery period. Altogether, this methodology opens the avenue for multi-timescale characterisation of brain states and provides an unprecedented window onto levels of vigilance.

Go/No-Go task engagement enhances population representation of target stimuli in primary auditory cortex.

Primary sensory cortices are classically considered to extract and represent stimulus features, while association and higher-order areas are thought to carry information about stimulus meaning. Here we show that this information can in fact be found in the neuronal population code of the primary auditory cortex (A1). A1 activity was recorded in awake ferrets while they either passively listened or actively discriminated stimuli in a range of Go/No-Go paradigms, with different sounds and reinforcements. Population-level dimensionality reduction techniques reveal that task engagement induces a shift in stimulus encoding from a sensory to a behaviorally driven representation that specifically enhances the target stimulus in all paradigms. This shift partly relies on task-engagement-induced changes in spontaneous activity. Altogether, we show that A1 population activity bears strong similarities to frontal cortex responses. These findings indicate that primary sensory cortices implement a crucial change in the structure of population activity to extract task-relevant information during behavior.

4-Hz oscillations synchronize prefrontal-amygdala circuits during fear behavior.

Fear expression relies on the coordinated activity of prefrontal and amygdala circuits, yet the mechanisms allowing long-range network synchronization during fear remain unknown. Using a combination of extracellular recordings, pharmacological and optogenetic manipulations, we found that freezing, a behavioral expression of fear, temporally coincided with the development of sustained, internally generated 4-Hz oscillations in prefrontal-amygdala circuits. 4-Hz oscillations predict freezing onset and offset and synchronize prefrontal-amygdala circuits. Optogenetic induction of prefrontal 4-Hz oscillations coordinates prefrontal-amygdala activity and elicits fear behavior. These results unravel a sustained oscillatory mechanism mediating prefrontal-amygdala coupling during fear behavior.