Publications using TDT

Non-rapid eye movement (NREM) sleep is not uniform but characterized by brief intrusions of wake-like brain activity and increased muscle tonus known as micro-arousals or brief awakenings. Although micro-arousals are an inherent feature of human and animal sleep, knowledge about the neural correlates of micro-arousals is sparse. We here developed an algorithm for automatic detection of micro-arousals based on EMG activity and used it to analyse the associated laminar neural activity in the motor cortex in mice. Our analysis showed that short micro-arousals with a duration below 5 s were associated with decreased cortical firing, while longer micro-arousals with a duration of 5–10 s were associated with increased firing in a similar manner to transitions to wakefulness. Analysis of single-channel firing showed that some channels exhibited increased activity immediately prior to micro-arousals, while others exhibited decreased activity. Slow wave activity (SWA, 1–4 Hz) immediately after micro-arousals was tightly correlated with sleep pressure and even surpassed average SWA levels during NREM sleep in sleep deprived animals. This study provides new insights into the neural mechanisms associated with micro-arousals and identifies a new link between sleep architecture and sleep homeostasis.

Animals synchronize their movements with external rhythms to coordinate perception and action, but the neural population mechanisms that allow them to attend and then initiate and sustain these rhythms remain unclear. Using recordings from the medial premotor cortex (MPC) of two macaque monkeys, we investigated neural dynamics during an attend‐then‐synchronize tapping task with visual metronomes. We found low‐dimensional neural manifolds that captured neural population trajectories. During the attention epoch, trajectories exhibited increasing amplitude and oscillatory strength with the successive stimuli, which is consistent with a dynamic system using a resonant mechanism. Then, the transition from perception to tapping synchronization was marked by a reliable shift into a distinct manifold subspace with rotatory dynamics, enabling accurate decoding of the switch in behavior. In addition, correct tapping trials were characterized by a more robust oscillatory structure not seen in incorrect trials. These findings demonstrate that the geometry and coherence of MPC neural trajectories encode different perceptual and motor aspects of tapping synchronization.

Noise exposure is a known cause of hearing loss, and only a few effective preventive drugs are available. Therefore, in this study, we aimed to investigate the protective effects of brimonidine on noise‐induced inner ear hearing impairment in mice and explore its underlying mechanisms and long‐term outcomes. Mice were randomly divided into control, noise exposure, and brimonidine groups. A 62‐week follow‐up was conducted after noise exposure. Brimonidine inhibited the noise‐induced increase in inner ear glutamate concentration and downregulated inflammatory factors and immunoglobulins. Brimonidine decreased glutaminase and VGLUT2/3 expression and reduced glutamate synthesis and vesicle transport without affecting its clearance, thereby decreasing glutamate excitotoxicity and protecting synapses and spiral ganglion neurons long term. Mice exposed to noise could temporarily restore their hearing thresholds; however, their auditory function in old age remained significantly worse than those that received brimonidine‐mediated cochlear protection in youth. These findings highlight the importance of enhancing noise protection from an early age.

Tissue inflammation or nerve injury at the periphery can cause chronic pain. Although the spinal-cord-projecting neurons in the rostral ventromedial medulla (RVMSC neurons) can promote pain chronification1-4, the pathway by which peripheral injury signals drive these neurons is poorly understood1-3,5. Here we report a circuit loop that extends from the spinal cord to the ventral posterolateral thalamus and posterior complex of the thalamus, proceeds to the primary somatosensory cortex and returns to the spinal cord via the lateral superior colliculus, which in turn connects to μ-opioid-receptor-expressing RVMSC neurons. Silencing any node along this multisynaptic circuit has minimal effects on nociception in healthy mice, but can eliminate mechanical hypersensitization and restore normal nociceptive response thresholds in mouse models of inflammatory and neuropathic pain. In healthy mice, repetitive-but not acute-activation of each node in this circuit is sufficient to cause robust chronic mechanical hypersensitization. Our findings reveal a spino-brain-spinal cord circuit loop that links ascending and descending pathways and specifically drives chronic mechanical pain. This could enable the identification of cellular targets for treating chronic pain.

Developing safe and effective pain medications is an ongoing challenge for human health. Agonists for the µ-opioid receptor (MOR) are essential pain medications, but their high intrinsic efficacy also induces adverse side effects, including respiratory depression, constipation, tolerance, dependence, withdrawal and addiction 1–7 . Strategies to limit adverse effects traditionally include developing MOR agonists that have low intrinsic efficacy or that preferentially activate G-protein signalling over β-arrestin signalling 8 . Here we identify a novel MOR agonist with supramaximal intrinsic efficacy and a unique pharmacological profile that produced effective analgesia in rodents with minimal adverse effects. N -desethyl-fluornitrazene (DFNZ) was derived from a class of synthetic benzimidazole opioids called nitazenes. DFNZ has impaired brain penetrance, a unique spatiotemporal MOR cellular signalling profile, and diminished efficacy at the MOR–galanin 1 receptor (GAL1) heteromer. DFNZ does not induce respiratory depression, tolerance or MOR downregulation after repeated exposure. Compared with other MOR agonists, DFNZ has limited effects on dopamine neurotransmission in nucleus accumbens and weaker reinforcing effects in the drug self-administration procedure. These results provide novel insights about MOR and nitazene pharmacology, have important implications for pain and addiction treatment, and challenge the prevailing dogma that high-efficacy MOR agonists cannot constitute safe and effective therapeutic agents.

Approximately 50% of hereditary deafness cases and most age-related hearing loss result from cochlear hair cell dysfunction. Adeno-associated virus (AAV)-mediated gene therapy offers a promising strategy for hearing restoration. However, the cochlea's complex cellular composition poses a significant challenge for targeted gene delivery into hair cells. Here, we establish a cross-species screening platform for cochlear AAV tools, applicable from rodents to large animals. Using this platform, we identify human promoters that specifically target inner hair cells, outer hair cells, or all hair cells in mice. The hair cell-specific coProB2 promoter is further validated in Bama miniature pigs and cynomolgus monkeys, achieving specific transgene expression in hair cells without auditory toxicity. Importantly, coProB2-driven gene therapies in DFNB79 and DFNB9 mouse models restore auditory function to near wild-type levels. Thus, this study establishes a cross-species-compatible, hair cell-specific AAV system, providing a versatile toolbox for cochlear hair cell gene therapy.

Glucose-dependent insulinotropic polypeptide (GIP) is a gut-derived incretin hormone, and pharmacologic modulation of central GIP receptors (GIPRs) improves energy homeostasis and prevents conditioned taste avoidance (CTA). However, the mechanisms by which GIPR signaling impact food intake and aversion are incompletely understood. Here, we show that GIPR agonism abrogates the aversive and enhances the anorexigenic effects of the pro-inflammatory cytokine interleukin-1β (IL-1β). Aversion-encoding parabrachial calcitonin gene-related peptide (CGRP) neurons were required for IL-1β-induced CTA but not anorexia. Moreover, systemic IL-1β increased CGRP neural activity in vivo, and this was significantly attenuated by co-administration of a GIPR agonist. By contrast, GIPR in the dorsal vagal complex was required for the acute anorectic effect of GIPR agonism but not its anti-aversive effect. Taken together, our data suggest that GIPR agonism reduces food intake and prevents aversion via distinct circuits and that GIPR agonism may represent an effective approach to alleviate inflammation-induced aversion.

Background: Genetic diagnosis plays a critical role in enhancing the identification and clinical management of pediatric hearing loss, ultimately improving both physiological outcomes and social-emotional development. As one of the most prevalent sensory disorders characterized by substantial genetic heterogeneity, hearing loss requires the implementation of efficient diagnostic approaches to inform targeted clinical interventions and personalized care strategies. Methods: Here, we recruited 3353 Han Chinese children with nonsyndromic hearing loss, including 598 multiplex cases with family history. We employed SNPscan, targeted panel sequencing (TPS), and whole-exome sequencing (WES) to delineate the mutation landscape. Findings: Utilizing a tiered genomic testing strategy, we performed SNPscan for all 3353 participants, TPS for 675 cases, and WES for 204 multiplex cases. We achieved an overall diagnostic yield of 48.3% (1620/3353) in this cohort. Our analysis identified 87 previously unreported variants across 24 genes, including a de novo FOXI1 mutation (c.479_481del) linked to Mondini malformation and enlarged vestibular aqueduct, validated through knock-in mouse models. Additionally, we discovered NEU4 as a candidate gene related to hearing loss, with knockout mice and cellular models demonstrating its role in auditory dysfunction via disrupted neuraminidase activity and axonal development. These findings enrich the spectrum of deafness-related genes and provide a new theoretical foundation for diagnosing hearing loss. Interpretation: This study not only expands the genetic spectrum of hearing loss but also underscores the clinical utility of advanced sequencing in guiding precision therapies and early syndromic surveillance. This study offers a detailed characterization of the genetic landscape of pathogenic variants associated with hearing loss in the Han Chinese population, which will contribute to the design of a cost-effective genetic screening scheme, and also provides valuable insights that contribute to the global understanding of the genetics of hearing loss. Funding: This work was supported by the Key Program of National Natural Science Foundation of China, the National Key Research and Development Program of China, the National Natural Science Foundation of China and the Major Fundamental Research Program of the Natural Science Foundation of Shandong Province, China.

Noise exposures causing transient hearing loss were previously considered benign. However, recent work has revealed that temporary noise-induced threshold shifts may be associated with long-lasting cochlear histopathology. One such effect is cochlear synaptopathy, i.e. changes to the afferent synapse between inner hair cells and auditory nerve fibers. Noise-induced synaptopathy has been extensively characterized in several rodent models, and temporal bone studies suggest similar age-related changes in humans. However, it remains unclear how noise-induced temporary threshold shifts affect cochlear structures in humans and nonhuman primates, which show greater resistance to noise exposure than other animals. Additionally, the long-term sequelae of temporary threshold shifts are largely unknown. Here, we characterized the effects of a noise exposure causing temporary threshold shifts on cochlear histopathology in macaque monkeys at long post-exposure survival times. Overall, cochlear histopathology was variable across subjects, similar to the variable susceptibility observed in humans. At 2 and 10 months post-exposure, macaques had no significant loss of hair cells, inner hair cell synapses, or cholinergic efferent innervation. However, both inner and outer hair cell ribbons showed significant enlargement. Together, these findings provide insight into the cochlear effects of single-exposure temporary threshold shifts in nonhuman primates.

Objective. Low-intensity transcranial focused ultrasound (tFUS) offers high spatial specificity and deep brain penetration, representing a promising non-invasive approach for modulating brain activity and behavior. Although emerging studies indicate that tFUS can modulate pain-related behaviors in rodents and humans, the underlying network-level and cellular mechanisms remain unknown. This study investigates the effects of tFUS neuromodulation on inhibitory neural circuits in a humanized mouse model of chronic pain, integrating electrophysiological, molecular, and histological analyses across a cohort of 50 animals, including wild-type controls. Approach. A 128-element random array transducer was used to precisely target the pain-processing brain circuit, while a non-invasive and flexible 30-channel electroencephalography electrode was applied to record local evoked responses, topographical activity, and global excitation and inhibition dynamics, which were further validated by optogenetics experiments. Cellular-level modulation and safety outcomes were evaluated through blinded histological examination. Main results. We found that tFUS produced robust modulation of local and global brain activity, characterized by suppression of local theta oscillations and enhancement of network-level inhibitory dynamics. These electrophysiological patterns aligned with those observed during optogenetic activation of parvalbumin (PV) interneurons. Immunohistochemistry further showed significant increases in inhibitory neuronal markers, including elevated expressions of glutamate decarboxylase 67 and PV. Blinded histological assessment confirmed the absence of tissue damage, supporting the safety of the stimulation paradigm. Significance. These findings demonstrated that tFUS stimulation non-invasively engages PV GABAergic inhibitory circuits in a chronic pain mouse model, providing mechanistic insight and supporting its development as a precise and safe neuromodulation technology for clinical translation.

Implantable biohybrid systems with computer-controlled actuation offer the capacity to modulate biological forces, but require biocompatible, self-sustaining, and scalable actuators. Engineering biological muscles can fulfill this need. However, muscle fatigue limits the fundamental capabilities of muscle-actuated systems. Here we present a fatigue-resistant myoneural actuator (MNA) with engineered recruitment biophysics in a rodent model. The MNA is based on manipulating native axonal composition through sensory reinnervation. This regenerative approach establishes functional neuromuscular junctions and redirects volitional control to computer control via nerve stimulation while maintaining self-sustainability. Compared to native muscles without the myoneural manipulation, fatigue resistance is augmented by 260%. Furthermore, we demonstrate closed-loop control with reversible neural isolation of the actuator, preventing unintended neural signaling to the central nervous system during operation. To illustrate the potential of the MNA technology, we present a biohybrid neuroprosthetic interface and a biohybrid organ system capable of modulating neural afferents and organ mechanics, respectively. Our framework demonstrates augmented biological muscle actuation while maintaining inherent tissue properties, bridging the technological gap for implantable biohybrid systems.

Age-related hearing loss (ARHL) is a rapidly growing public health concern, affecting two-thirds of adults over 65 years old, with no effective therapeutics available. As the aging population grows at an unprecedented rate, the burden of ARHL will only increase. The causes of ARHL are multifactorial, but an understudied major contributor is glial dysfunction. The auditory nerve (AN) conducts sound from the cochlea to the brainstem and holds a diverse population of immune cells and myelinating glia. As the AN fibers bundle together within the cochlea to project to the brainstem, they are first myelinated by Schwann cells in the peripheral AN, then myelinated by oligodendrocytes in the central AN. The region where myelination shifts from Schwann cells to oligodendrocytes is the glial transition zone (GTZ), located in the cochlear modiolus, creating a unique biological niche. While central-peripheral interfaces are recognized in other cranial nerves, the AN GTZ is understudied. This region integrates the peripheral and central microenvironments within the confined bony cochlea, positioning it as a niche for glial dysfunction in pathological conditions, such as aging. We hypothesize that the GTZ is a site of enhanced glial dysfunction contributing to age-related AN demyelination, an important contributor to ARHL. We evaluated this in an ARHL mouse model combining RNA-sequencing, quantitative immunohistochemistry, and 3D high-resolution imaging. We examined the AN GTZ from human temporal bone donors. RNA-sequencing of the AN revealed age-associated increases in abnormal myelination/glial function and inflammation. There was a significant age-dependent increase in Iba1 + macrophages/microglia, with accumulation at the AN GTZ, and an increase in cellular volume and surface area, suggesting greater age-related activation. Macrophages/microglia contained significantly more internalized myelin debris in the AN (peripheral, central, and GTZ) with aging. More importantly, we found structurally intact myelin within macrophages/microglia only at the GTZ, suggesting a unique microenvironment at the GTZ altering phagocytic activity in aging. Together, our data suggest that the GTZ, a previously unrecognized central-peripheral interface, is a critical site of immune-glial interactions and especially vulnerable to age-related demyelination and neuroinflammation. This study highlights the GTZ as a potential target for preserving AN myelination and mitigating ARHL.

Aminoglycoside antibiotics remain crucial for the treatment of severe infections caused by multidrug-resistant bacteria; however, their clinical use is limited by the risk of irreversible hearing loss. Previous studies have primarily attributed aminoglycoside-induced hearing loss to cochlear hair cell loss and subsequent spiral ganglion degeneration, while the contribution of the stria vascularis remains unclear. In the present study, we demonstrate that gentamicin administration induces apoptosis of stria vascularis pericytes, leading to stria vascularis dysfunction and consequent hearing loss in mice. Mechanistically, gentamicin exposure activated macrophages within the stria vascularis and significantly increased local expression of tumor necrosis factor-α (TNF-α). In vitro, we found that TNF-α triggered stria vascularis pericyte apoptosis via activation of the NF-κB signaling pathway. Importantly, in vivo pharmacological blockade of TNF-α with infliximab (IFX) or inhibition of NF-κB signaling with JSH-23 effectively preserved pericyte survival, attenuated stria vascularis damage, and mitigated gentamicin-induced hearing loss in mice. Collectively, these findings uncover TNF-α/NF-κB-dependent vascular-inflammatory mechanism underlying gentamicin ototoxicity and identify stria vascularis pericytes as a novel and druggable therapeutic target for the prevention of gentamicin-induced hearing loss.

Endocannabinoid (eCB) signaling is a key regulator of reward-related dopaminergic signaling, particularly in response to drugs of abuse, such as cocaine. To date, our understanding of this mechanism has primarily been limited to male subjects. Prior work establishes that female cocaine users have more adverse outcomes, and female rats show greater sensitivity to cannabinoid type 1 receptor (CB1R) regulation of cocaine self-administration. Therefore, we hypothesize that female rats exhibit enhanced eCB regulation of cocaine-evoked dopamine (DA). We used in vivo fiber photometry recording of the dopamine biosensor, dLight 1.3b, in the nucleus accumbens medial shell (NAc ms ) in response to cocaine in male and female rats. Rats were pretreated with cannabinoid-targeting drugs to investigate the effects of CB1R inactivation or augmentation of the eCB 2-AG on cocaine-evoked DA. Our results revealed that CB1R inactivation attenuates cocaine-evoked DA in male and female rats, but females showed enhanced sensitivity for CB1R regulation of cocaine-evoked DA. Cocaine-evoked DA was enhanced by augmenting 2-AG levels, and females again showed increased sensitivity to this manipulation. Finally, females show greater cocaine-evoked DA when in a non-estrous cycle compared to estrous, reinforcing that estrous cycle is a determinant of cocaine-evoked DA. These data indicate that females show enhanced eCB regulation of cocaine-evoked DA signaling, underscoring the importance of sex as a biological variable in our understanding of endocannabinoid regulation of drug reward. Highlights CB1R inactivation attenuates cocaine-evoked DA in NAc ms , preferentially in females 2-AG augmentation via MAGL inhibition enhances cocaine-evoked DA, with female bias Estrous phase modulates the dopamine response to a high dose of cocaine in females Male and female rats show similar baseline DA and locomotor responses to cocaine

Age-related hearing loss (ARHL), or presbycusis, is one of the most prevalent sensory deficits in older adults and has been increasingly implicated in cognitive decline and dementia. This study characterizes ARHL in a rhesus macaque model by combining histological, physiological, and cognitive assessments. Aged macaques exhibited progressive cochlear degeneration, with marked outer hair cell loss at mid-to-high frequencies, elevated auditory thresholds, reduced distortion product otoacoustic emissions, and impaired auditory brainstem responses including amplitude reduction, latency prolongation, and diminished temporal precision. Despite modest reductions in inner hair cell ribbon synapse counts, hypertrophic changes were observed. These auditory deficits correlated with subtle impairments in visual working memory, as measured by a delayed match-to-sample task, underscoring a potential sensory-cognitive link. By capturing cross-domain aging markers in a translationally relevant primate model, this work lays a foundation for mechanistic studies and therapeutic interventions targeting both hearing and cognition in aging populations.

Background Peripheral hearing loss is associated with auditory hallucinations and increased risk for psychotic disorder, particularly in genetically vulnerable individuals. Moreover, both hearing loss and schizophrenia disrupt auditory temporal acuity, a sensitive measure of auditory brain function. Here, we used mouse models of hearing loss and schizophrenia to reveal how neural mechanisms of auditory cortical temporal acuity depend on sensory and genetic risk factors for psychosis. Methods We quantified auditory cortical temporal acuity in mice with or without hearing loss and with or without the 22q11.2 deletion — one of the strongest known genetic risk factors for schizophrenia. Cortical single-unit and population activity were recorded in awake mice (N = 23) during presentations of loudness-adjusted gap-in-noise stimuli with varying durations of silent gap, which are commonly used to assess auditory temporal acuity in humans. Results Both hearing loss and the 22q11.2 deletion disrupted auditory cortical temporal acuity, but through distinct mechanisms. Hearing loss broadly degraded temporal acuity at the level of single-unit responses and neural population activity. In contrast, the 22q11.2 deletion selectively impaired gap duration thresholds in regular-spiking (putative excitatory) but not fast-spiking (putative inhibitory) neurons. Mice with comorbid hearing loss and 22q11.2 deletion exhibited both abnormalities in auditory cortical temporal acuity. Conclusions Sensory and genetic risk factors for psychosis can disrupt auditory cortical temporal acuity via distinct mechanisms that remain partially dissociable even under comorbid conditions. These findings underscore the importance of accounting for hearing loss comorbidity when interpreting auditory cortical dysfunction in psychotic disorder.

The development of motor control in primary motor cortex (M1) requires both movements and neural activity. In rats, cortical motor control first appears around postnatal day (P) 25, prior to which movements are generated by subcortical motor nuclei such as the red nucleus (RN). While these subcortically generated movements are thought to provide the activity that guides the development of M1, the specific movements associated with M1 activity and their somatotopic and temporal precision remain unknown. Here, we performed acute electrophysiological recordings of neural activity in the forelimb region of M1 of P12–24 rats as they cycled between sleep and wake and compared M1 and RN activity in P24 rats. At every age, M1 neurons exhibited somatopically precise activity during REM sleep twitches, along with strong activity during wake movements. From P12 to P24, the proportion of neurons exhibiting twitch-related activity decreased, twitch-related activity became more temporally refined, and a larger fraction of spikes occurred before movement onset. At P24, M1 showed less premovement activity than RN. Further, in contrast to the non-selective activity seen in M1, some RN neurons showed movement-selective activity patterns during wake, firing only during particular wake movements. These findings reveal that movement-related activity in M1 is somatotopically precise by P12 and temporally precise by P24. But M1 still lacks the strong premovement activity and selectivity characteristic of RN, suggesting that at P24, subcortical outputs remain the main drivers of M1’s movement-related activity.

Male bushcrickets produce complex species-specific songs to attract mating partners. Since the neuronal processing underlying song-pattern detection is not fully understood, we analysed extracellularly recorded song-induced spike responses and local field potentials across the first stage of auditory processing in the prothoracic ganglion of Mecopoda elongata. We analysed the influence of the temporal structure and of any change in amplitude of spiking and local field potentials. We presented the complete chirp and compared the neuronal response pattern with those responses evoked individually by each pulse of the chirp. Our data revealed: (i) the complete chirp leads to a transient summation in local field potentials over the time course of the chirp, and (ii) significantly fewer action potentials occur during the chirp presentation than to the individually presented pulses. Our data provide insights into the question of how an acoustical signal can be used to extract those features from sensory information that are important for auditory behaviour, such as chorus formation, species recognition, or phonotaxis.

Male parenting behavior is highly conserved and activated only after offspring are born. Here we show that the behavioral switch from infanticide in virgin male mice to caregiving in wild-type sires is associated with enhanced expression and activation of Transient receptor potential channel 5 (Trpc5) in estrogen receptor α (Esr1)-expressing neurons in the hypothalamic medial preoptic area (MPOA). While selective deletion of Trpc5 from Esr1 MPOA neurons diminishes paternal behavior in sires, overexpression of Trpc5 in these neurons converts the otherwise infanticidal virgin males to exhibit care to pups. Mechanistically, Trpc5-dependent changes in neuronal excitability underlie fatherhood-associated Esr1 MPOA neuron plasticity. Notably, Trpc5 overexpression also enhances escape behavior and elicits exploratory diving behavior, suggesting that these neurons control a broader adaptive parenting response in males. These findings establish a Trpc5-dependent MPOA neural signal as a critical regulator of paternal behavior.

Every moment, a spotlight of attention roams our visual field to enhance detection of salient stimuli. While recent work suggests how the brain selects salient relevant stimuli (attentional focus), it is unknown if there are mechanisms that modulate attention to suit fluctuating target salience or internal needs (attentional strength). Here, using optical recordings and perturbations in mice performing a cued detection task, we report that rapid dynamics in both dorsal raphe (DR) activity and cortical serotonin (5HT) release selectively regulate attentional strength. In contrast to these rapid DR-5HT dynamics, we found previously reported reward-associated and slower dynamics were both unrelated to attention. Brief optogenetic DR-5HT suppression increased attentional strength and performance, whereas optogenetic excitation reduced attentional strength and performance. A computational model suggests DR-5HT’s inverse effects on attention arise from control of cortical suppression and divisive normalization. Collectively, our results demonstrate that the brain independently controls attentional strength and focus and defines rapid DR-5HT neuromodulation as a new regulator of attentional strength.