OBJECTIVE: Absence seizures impair psychosocial function, yet their detailed neuronal basis remains unknown. Recent work in a rat model suggests that cortical arousal state changes prior to seizures and that single neurons show diverse firing patterns during seizures. Our aim was to extend these investigations to a mouse model with studies of neuronal activity and arousal state to facilitate future fundamental investigations of absence epilepsy.
METHODS: We performed in vivo extracellular single unit recordings on awake head-fixed C3H/HeJ mice. Mice were implanted with tripolar electrodes for cortical electroencephalography (EEG). Extracellular single unit recordings were obtained with glass micropipettes in the somatosensory barrel cortex, while animals ambulated freely on a running wheel. Signals were digitized and analyzed during seizures and at baseline.
RESULTS: Neuronal activity was recorded from 36 cortical neurons in 19 mice while EEG showed characteristic 7-8 Hz spike-wave discharges. Different single neurons showed distinct firing patterns during seizures, but the overall mean population neuronal firing rate during seizures was no different from pre-seizure baseline. However, the rhythmicity of neuronal firing during seizures was significantly increased (p < 0.001). In addition, beginning 10s prior to seizure initiation, we observed a progressive decrease in cortical high frequency (>40 Hz) EEG and an increase in lower frequency (1-39 Hz) activity suggesting decreased arousal state.
CONCLUSIONS: We found that the awake head-fixed C3H/HeJ mouse model demonstrated rhythmic neuronal firing during seizures, and a decreased cortical arousal state prior to seizure onset. Unlike the rat model we did not observe an overall decrease in neuronal firing during seizures. Similarities and differences across species strengthen the ability to investigate fundamental key mechanisms. Future work in the mouse model will identify the molecular basis of neurons with different firing patterns, their role in seizure initiation and behavioral deficits, with ultimate translation to human absence epilepsy.

Age is a primary risk factor for Parkinson\'s disease (PD); however, the effects of aging on the Parkinsonian brain remain poorly understood, particularly for deep brain structures. We investigated intraoperative micro-electrode recordings from the subthalamic nucleus (STN) of PD patients aged between 42 and 76 years. Age was associated with decreased oscillatory beta power and non-oscillatory high-frequency power, independent of PD-related variables. Single unit firing and burst rates were also reduced, whereas the coefficient of variation and the structure of burst activity were unchanged. Phase synchronization (debiased weighed phase lag index [dWPLI]) between sites was pronounced in the beta band between electrodes in the superficial STN but was unaffected by age. Our results show that aging is associated with reduced neuronal activity without changes to its temporal structure. We speculate that the loss of activity in the STN may mediate the relationship between PD and age.

Vocalization, a means of social communication, is prevalent among many species, including humans. Both rats and mice use ultrasonic vocalizations (USVs) in various social contexts and affective states. The motor cortex is hypothesized to be involved in precisely controlling USVs through connections with critical regions of the brain for vocalization, such as the periaqueductal gray matter (PAG). However, it is unclear how neurons in the motor cortex are modulated during USVs. Moreover, the relationship between USV modulation of neurons and anatomical connections from the motor cortex to PAG is also not clearly understood. In this study, we first characterized the activity patterns of neurons in the primary and secondary motor cortices during emission of USVs in rats using large-scale electrophysiological recordings. We also examined the axonal projection of the motor cortex to PAG using retrograde labeling and identified two clusters of PAG-projecting neurons in the anterior and posterior parts of the motor cortex. The neural activity patterns around the emission of USVs differed between the anterior and posterior regions, which were divided based on the distribution of PAG-projecting neurons in the motor cortex. Furthermore, using optogenetic tagging, we recorded the USV modulation of PAG-projecting neurons in the posterior part of the motor cortex and found that they showed predominantly sustained excitatory responses during USVs. These results contribute to our understanding of the involvement of the motor cortex in the generation of USV at the neuronal and circuit levels.

Numerous studies have suggested that N-acetylcysteine (NAC), has the potential to suppress drug craving in people with substance use disorder and reduce drug-seeking behaviors in animals. The nucleus accumbens (NAc) plays a crucial role in the brain\'s reward system, with the nucleus accumbens core (NAcore) specifically implicated in compulsive drug seeking and relapse. In this study, we aimed to explore the impact of subchronic NAC administration during the extinction period and acute NAC administration on the electrical activity of NAcore neurons in response to a priming dose of morphine in rats subjected to extinction from morphine-induced place preference (CPP).We conducted single-unit recordings in anesthetized rats on the reinstatement day, following the establishment of morphine-induced conditioned place preference (7 mg/kg, s.c., 3 days), and subsequent drug-free extinction. In the subchronically NAC-treated groups, rats received daily injections of either NAC (50 mg/kg; i.p.) or saline during the extinction period. On the reinstatement day, we recorded the spontaneous activity of NAcore neurons for 15 min, administered a priming dose of morphine, and continued recording for an additional 45 min. While morphine excited most recorded neurons in saline-treated rats, it failed to alter firing rates in NAC-treated rats that had received NAC during the extinction period. For acutely NAC-treated animals, we recorded the baseline activity of NAcore neurons for 10 min before administering a single injection of either NAC (50 mg/kg; i.p.) or saline in rats with no treatment during the extinction. Following 30 min of recording and a priming dose of morphine (1 mg/kg, s.c.), the recording continued for an additional 30 min. The firing activity of NAcore neurons did not show significant changes after morphine or NAC injection. In conclusion, our findings emphasize that daily NAC administration during the extinction period significantly attenuates the morphine-induced increase in firing rates of NAcore neurons during the reinstatement of morphine CPP. However, acute NAC injection does not produce the same effect. These results suggest that modulating glutamate transmission through daily NAC during extinction may effectively inhibit the morphine place preference following the excitatory effects of morphine on NAcore neurons.

UNASSIGNED: Despite 6%-20% of the adult population suffering from tinnitus, there is no standard treatment for it. Placenta extract has been used for various therapeutic purposes, including hearing loss. Here, we evaluate the effect of a novel neuroprotective protein composition (NPPC) extract on electrophysiological and molecular changes in the medial geniculate body (MGB) of tinnitus-induced rats.
UNASSIGNED: To evaluate the protein analysis by western blot, the rats were divided into three groups: (1) saline group (intraperitoneal injection of 200 mg/kg saline twice a day for 28 consecutive days, (2) chronic Na-Sal group received sodium salicylate as in the first group, and (3) chronic treatment group (received salicylate 200 mg/kg twice daily for 2 weeks, followed by 0.4 mg NPPC daily from day 14 to day 28). Single-unit recordings were performed on a separate group that was treated as in group 4. Gap-prepulse inhibition of the acoustic startle (GPIAS) and pre-pulse inhibition (PPI) was performed to confirm tinnitus in all groups at the baseline, 14th and 28th days.
UNASSIGNED: Western blot analysis showed that the expression of γ-Aminobutyric acid Aα1 subunit (GABA Aα1), N-methyl-d-aspartate receptor subtype 2B (NR2B or NMDAR2B), α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors subunit GluR1 (GluR1), and α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors subunit GluR2 (GluR2) decreased after Na-Sal injection, while NPPC upregulated their expression. MGB units in rats with tinnitus showed decreased spontaneous firing rate, burst per minute, and a spike in a burst. After NPPC administration, neural activity patterns showed a significant positive effect of NPPC on tinnitus.
UNASSIGNED: NPPC can play an effective role in the treatment of tinnitus in salicylate-induced rats, and MGB is one of the brain areas involved in these processes.
UNASSIGNED: NA.

Behavioral neuroscience has long relied on in vivo electrophysiology to provide spatially and temporally precise answers to complex questions about the neural dynamics underlying sensory processing and action execution. Investigating the neural correlates of behavior can be challenging in freely behaving animals, especially when making inferences related to internal states that are temporally or conceptually ambiguous, such as decision-making or motivation. This necessitates careful creation of appropriate and rigorous controls and awareness of the many potential confounds when attributing neural signals to animal behavior. This article discusses fundamental considerations for the optimal design and interpretation of in vivo rodent electrophysiological recording experiments and focuses on the different optimization strategies required when investigating neural encoding of external stimuli versus free behavior. The first protocol offers suggestions specific to intracranial surgical implantation of multielectrode arrays. The second protocol delves into optimization strategies and tips useful for designing and interpreting recording experiments conducted in freely behaving rodents. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Surgical implantation of the multielectrode array Basic Protocol 2: Optimizing experimental design and parameters.

There are vast gaps in our understanding of the organization and operation of the human nervous system at the level of individual neurons and their networks. Here, we report reliable and robust acute multichannel recordings using planar microelectrode arrays (MEAs) implanted intracortically in awake brain surgery with open craniotomies that grant access to large parts of the cortical hemisphere. We obtained high-quality extracellular neuronal activity at the microcircuit, local field potential level and at the cellular, single-unit level. Recording from the parietal association cortex, a region rarely explored in human single-unit studies, we demonstrate applications on these complementary spatial scales and describe traveling waves of oscillatory activity as well as single-neuron and neuronal population responses during numerical cognition, including operations with uniquely human number symbols. Intraoperative MEA recordings are practicable and can be scaled up to explore cellular and microcircuit mechanisms of a wide range of human brain functions.

Current sub-perception spinal cord stimulation (SCS) is characterized by the use of high-frequency pulses to achieve paresthesia-free analgesic effects. High-frequency SCS demonstrates distinctive properties from paresthesia-based SCS, such as a longer time course to response, implying the existence of alternative mechanism(s) of action beyond gate control theory. We quantified the responses to SCS of single neurons within the superficial dorsal horn (SDH), a structure in close proximity to SCS electrodes, to investigate the mechanisms underlying high-frequency SCS in 62 urethane-anesthetized male rats. Sciatic nerve stimulation was delivered to isolate lumbar SDH neurons with evoked C-fiber activity. The evoked C-fiber activity before and after the application of SCS was compared to quantify the effects of SCS across stimulation intensity and stimulation duration at three different stimulation frequencies. We observed heterogeneous responses of SDH neurons which depended primarily on the type of unit. Low-threshold units with spontaneous activity, putatively inhibitory interneurons, tended to be facilitated by SCS while the other unit types were suppressed. The effects of SCS were more prominent with increased stimulation duration from 30 s to 30 m across frequencies. Our results highlight the importance of inhibitory interneurons in modulating local circuits of the SDH and the importance of local circuit contributions to the analgesic mechanisms of SCS.

This study was conducted in order to reveal the possibly lateralized processes in the avian nidopallium caudolaterale (NCL), a functional analogue to the mammalian prefrontal cortex, during a color discrimination task. Pigeons are known to be visually lateralized with a superiority of the left hemisphere/right eye for visual feature discriminations. While animals were working on a color discrimination task, we recorded single visuomotor neurons in left and right NCL. As expected, pigeons learned faster and responded more quickly when seeing the stimuli with their right eyes. Our electrophysiological recordings discovered several neuronal properties of NCL neurons that possibly contributed to this behavioral asymmetry. We found that the speed of stimulus encoding was identical between left and right NCL but action generation was different. Here, most left hemispheric NCL neurons reached their peak activities shortly before response execution. In contrast, the majority of right hemispheric neurons lagged behind and came too late to control the response. Thus, the left NCL dominated the animals\' behavior not by a higher efficacy of encoding, but by being faster in monopolizing the operant response. A further asymmetry concerned the hemisphere-specific integration of input from the contra- and ipsilateral eye. The left NCL was able to integrate and process visual input from the ipsilateral eye to a higher degree and thus achieved a more bilateral representation of two visual fields. We combine these novel findings with those from previous publications to come up with a working hypothesis that could explain how hemispheric asymmetries for visual feature discrimination in birds are realized by a sequential buildup of lateralized neuronal response properties in the avian forebrain.

Of all frontocortical subregions, the anterior cingulate cortex (ACC) has perhaps the most overlapping theories of function.1-3 Recording studies in rats, humans, and other primates have reported diverse neural responses that support many theories,4-12 yet nearly all these studies have in common tasks in which one event reliably predicts another. This leaves open the possibility that ACC represents associative pairing of events, independent of their overt biological significance. Sensory preconditioning13 provides an opportunity to test this. In the first phase, preconditioning, value-neutral sensory stimuli are paired (A→B). To test whether this was learned, subjects are given standard conditioning during which one of the previously neutral sensory cues is paired with a biologically meaningful outcome (B→outcome). During the final probe test, the neutral cue which was never paired with a biologically meaningful outcome is presented alone (A→) and will elicit a conditional response, suggesting that subjects had learned the associative structure during preconditioning and use that knowledge to infer presentation of the biologically relevant outcome (A→B→outcome). Inference-based responding demonstrates a fundamental property of model-based reasoning14,15 and requires learning of the associations between neutral stimuli before rewards are introduced.16-19 ACC neurons developed firing patterns that reflected the learning of sensory associations during preconditioning, even though no rewards were present. The strength of these correlates predicted rats\' ability to later mobilize and use that associative information during the probe test. These results demonstrate that clear biological significance is not necessary to produce correlates of learning in ACC.