Changes In Neuronal Activity Research Articles

Weak representation of awake/sleep states by local field potentials in aged mice

Senescence affects various aspects of sleep, and it remains unclear how sleep-related neuronal network activity is altered by senescence. Here, we recorded local field potential signals from multiple brain regions covering the forebrain in young (10-week-old) and aged (2-year-old) mice. Interregional LFP correlations across these brain regions could not detect pronounced differences between awake and sleep states in both young and aged mice. Multivariate analyses with machine learning algorithms with uniform manifold approximation and projection and robust continuous clustering demonstrated that LFP correlational patterns at multiple frequency bands, ranging from delta to high gamma bands, in aged mice less represented awake/sleep states than those in young mice. By housing aged mice in an enriched environment, the LFP patterns were changed to more precisely represent awake/sleep states. Our results demonstrate senescence-induced changes in neuronal activity at the network level and provide insight into the prevention of pathological symptoms associated with sleep disturbance in senescence.

Low-Intensity Transcranial Ultrasound Stimulation: Mechanisms of Action and Rationale for Future Applications in Movement Disorders.

Low-intensity transcranial ultrasound stimulation (TUS) is a novel non-invasive brain stimulation technique that uses acoustic energy to induce changes in neuronal activity. However, although low-intensity TUS is a promising neuromodulation tool, it has been poorly studied as compared to other methods, i.e., transcranial magnetic and electrical stimulation. In this article, we first focus on experimental studies in animals and humans aimed at explaining its mechanisms of action. We then highlight possible applications of TUS in movement disorders, particularly in patients with parkinsonism, dystonia, and tremor. Finally, we highlight the knowledge gaps and possible limitations that currently limit potential TUS applications in movement disorders. Clarifying the potential role of TUS in movement disorders may further promote studies with therapeutic perspectives in this field.

Age-Dependent FOSB/ΔFOSB Response to Acute and Chronic Stress in the Extended Amygdala, Hypothalamic Paraventricular, Habenular, Centrally-Projecting Edinger-Westphal, and Dorsal Raphe Nuclei in Male Rats.

FOS proteins are early-responding gene products that contribute to the formation of activator protein-1. Several acute and chronic stimuli lead to Fos gene expression, accompanied by an increase of nuclear FOS, which appears to decline with aging. FOSB is another marker to detect acute cellular response, while ΔFOSB mirrors long-lasting changes in neuronal activity upon chronic stress. The notion that the occurrence of stress-related mood disorders shows some age dependence suggests that the brain’s stress sensitivity is also a function of age. To study age-dependent stress vulnerability at the immediate-early gene level, we aimed to describe how the course of aging affects the neural responses of FOSB/ΔFOSB in the acute restraint stress (ARS), and chronic variable mild stress (CVMS) in male rats. Fourteen brain areas [central, medial, basolateral (BLA) amygdala; dorsolateral- (BNSTdl), oval- (BNSTov), dorsomedial-, ventral- (BNSTv), and fusiform- (BNSTfu) divisions of the bed nucleus of the stria terminalis; medial and lateral habenula, hypothalamic paraventricular nucleus (PVN), centrally-projecting Edinger-Westphal nucleus, dorsal raphe nucleus, barrel field of somatosensory cortex (S1)] were examined in the course of aging. Eight age groups [1-month-old (M), 1.5 M, 2 M, 3 M, 6 M, 12 M, 18 M, and 24 M] of rats were exposed to a single ARS vs. controls. In addition, rats in six age groups (2, 3, 6, 12, 18, and 24 M) were subjected to CVMS. The FOSB/ΔFOSB immunoreactivity (IR) was a function of age in both controls, ARS- and CVMS-exposed rats. ARS increased the FOSB/ΔFOSB in all nuclei (except in BLA), but only BNSTfu, BNSTv, and PVN reacted throughout the examined lifespan. The CVMS did not increase the FOSB/ΔFOSB in BLA, BNSTov, BNSTdl, and S1. PVN showed a constantly maintained FOSB/ΔFOSB IR during the examined life period. The maximum stress-evoked FOSB/ΔFOSB signal was detected at 2–3 M periods in the ARS- and at 6 M, 18 M in CVMS- model. Corresponding to our previous observations on FOS, the FOSB/ΔFOSB response to stress decreased with age in most of the examined nuclei. Only the PVN exerted a sustained age-independent FOSB/ΔFOSB, which may reflect the long-lasting adaptation response and plasticity of neurons that maintain the hypothalamus-pituitary-adrenal axis response throughout the lifespan.

Orexin A enhances neuronal synchronization in adult rat hypothalamic culture: A model to study hypothalamic function

BackgroundThe regulation of sleep/wake behavior and energy homeostasis is maintained in part by the lateral hypothalamic (LH) neuropeptide orexin A (OXA, hypocretin). A reduction in orexin signaling is associated with sleep disorders and obesity, whereas higher LH orexin signaling and sensitivity promotes obesity resistance. Similarly, dysregulation of hypothalamic neural networks is associated with onset of age‐related diseases, including obesity and several neurological diseases. Despite the increase in obesity with aging, and that adult populations are the subjects in the majority of pharmaceutical and obesity studies, conventional models for neuronal networks utilize embryonic neural cultures rather than adult neurons. Synchronous activity, a feature of normal brain function, and a measure of functional connectivity, describes correlated changes in neuronal activity between neurons, Synchronization determines the nature of the final output from a given neural structure. Neural synchrony is altered by behavioral perturbations, in embryonic neurons obtained from obesity‐resistant rats, and following application of OXA onto embryonic hypothalamic cultures. Synchronous network dynamics in adult hypothalamic neurons remain largely undescribed.MethodsWe established an adult rat hypothalamic culture in multi‐electrode‐array (MEA) dishes and recorded the field potentials. Then we studied the effect of exogenous OXA on network synchronization of these adult hypothalamic cultures. In addition, we studied the wake promoting effects of OXA in vivo when directly injected into the rostral lateral hypothalamus (rLH). To study the wake promoting effects of LH OXA, rats were implanted with a radiotelemetric transmitter and EEG/EMG electrodes to record vigilance states (F40‐EET, Data Sciences International [DSI], St. Paul, MN), and a cannula targeting the rLH. Following surgical recovery, either OXA (250 pmol/0.5 μl) or saline was infused into the rLH (single injection) and sleep/wake states were measured for 2h post‐injection.ResultsThe results show that adult hypothalamic cultures are viable for nearly 3 months in vitro, good quality MEA recordings can be obtained from these cultures in vitro, and finally, that cultured adult hypothalamus is responsive to OXA. In addition, LH administration of OXA enhanced wakefulness in rats, indicating that OXA enhances wakefulness for up to 2h post‐injection.ConclusionsThese results support that adult rat hypothalamic cultures could be used as a model to study the neural mechanisms underlying obesity, and that obesity resistance is associated with increased hypothalamus synchronization. Increased wake following rLH OXA suggest the possibility that OXA promotes wake partly by promoting neural synchrony in the hypothalamus.

Hypoxia Augments TRPM3‐mediated Calcium Influx in Vagal Sensory Neurons

Exposure to low systemic oxygen (hypoxia, Hx) elevates blood pressure, heart rate and respiration via reflex sensory and central nervous system activation. We have shown transient receptor potential (TRP) vanilloid channels contribute to afferent‐evoked activation of the brainstem nucleus tractus solitarii (nTS) following chronic Hx. Short sustained Hx exposure (10% O2, 24hr) in rats also enhances nTS activity, although the mechanisms are not completely understood. Recent evidence from others shows TRP melastatin 3 (TRPM3) channels contribute to afferent activity and nTS glutamate signaling, but their role in afferent signaling in Hx is unknown. We hypothesize that TRPM3 increases afferent neuronal activity in nTS‐projecting nodose ganglia (NG) neurons, and its contribution is elevated following Hx. Rats were exposed to either in vivo normal room air (normoxia) or 24‐hr of 10% O2 (Hx). A subset of NG neurons from normoxic rats were exposed to in vitro incubation for 24‐hr in 21% or 1% O2, 5% CO2. Intracellular calcium (Ca2+) of dissociated NG neurons was monitored via Fura‐2 fluorescence imaging. TRPM3 was activated via 1‐min bath application of Pregnenolone sulfate (Preg, 30 and 100 µM) or CIM0216 (5 ‐ 20 µM). The increase in cellular fluorescence to Preg or CIM0216 was normalized to that of its prior baseline fluorescence (ΔF/F%). NG neurons were also exposed to vehicle (Veh) alone and their Ca2+ ΔF/F% responses were used as the threshold for TRPM3 responders under comparable timeframes. RNAScope identified TRPM3 transcripts in the normoxic NG. Veh alone did not alter Ca2+ from their initial baseline (ΔF/F%, less than 102.5% increase across all preparations). Activation of TRPM3 via Preg (30 and 100 µM) elevated Ca2+ in 51% and 55% of NG neurons by a similar magnitude [120.4 ± 3.8% and 116.6 ± 3.0% (mean ± SEM), respectively]. CIM0216 also augmented Ca2+ with the greatest response occurring during 20 µM (~56% of NG neurons, 115.6 ± 3.1% elevation). Preg responses were significantly reduced by pre‐application of the TRPM3 antagonist ononetin (20 µM; Preg, 125.9 ± 7.7% vs. ononetin + Preg, 101.7 ± 0.6%), confirming drug specificity. Elevated Ca2+ by 30 µM Preg was also eliminated by bathing cells in 0 mM Ca2+ (reduced to 100.9 ± 0.4%) suggesting Ca2+ elevation occurs via opening of membrane‐bound TRPM3. In NG neurons isolated from Hx‐exposed rats, the Preg‐induced elevation of Ca2+ was significantly greater than their paired normoxic‐exposed rats (134.1 ± 4.2% vs 118.1 ± 3.9%), although the number of responders were comparable (40% vs 43%). To determine if the increase in TRPM3 Ca2+ responses to in vivo Hx was due to the reduction of oxygen or the reflex‐induced change in neuronal activity, cells were isolated then incubated in either 21% or 1% O2 for 24 hrs. Following 24 hr in vitro Hx, 30 µM Preg enhanced Ca2+ more than in 24 hr normoxic controls (115.3 ± 2.2% vs 108.1 ± 0.1%). The proportion of Preg responders was similar between groups (58% vs 61%). Altogether, these results suggest a Hx specific increase in TRPM3 function, driven by the influx of extracellular calcium.

Fadu head and neck squamous cell carcinoma induces hyperexcitability of primary sensory neurons in an in vitro coculture model.

Currently, cancer pain is viewed as a process orchestrated by the release of pronociceptive molecules and the invasion of neural structures, referred to as perineural invasion (PNI). Cancer pain resulting from PNI is well-documented, but the mechanisms leading to peripheral sensitization because of tumor growth are not fully known. A retrospective study was used to examine how the use of anti-inflammatory medications affected preoperative pain in patients with oral squamous cell carcinoma cancer. We then used an in vitro coculture model in which dorsal root ganglion (DRG) neurons were incubated together with Fadu human head and neck squamous cell carcinoma cancer cells to explore how cancer cells affect the electrical membrane properties of sensory neurons. We found that inflammation contributes to preoperative pain in patients with oral squamous cell carcinoma. After coculture with Fadu human head and neck squamous cell carcinoma cancer cells, we identified markers of inflammation in coculture media and found evidence of neuronal sensitization, including spontaneous activity, reduced current thresholds, depolarized resting membrane potential, and enhanced responses to current stimulation in human and rat DRG neurons. In rats, these effects were influenced by sex and age: neurons from young adult female rats were resistant to changes in neuronal activity, in contrast to neurons from older adult female rats or male rats of either age group. Pro-inflammatory substances released in cancer cell-DRG coculture promoted neuronal hyperexcitability and may contribute to cancer pain after PNI, and these effects may differ across age groups and sexes.

Molecular and cellular mechanisms for differential effects of chronic social isolation stress in males and females.

Chronic social isolation stress during adolescence induces susceptibility for neuropsychiatric disorders. Here we show that 5-week post-weaning isolation stress induces sex-specific behavioral abnormalities and neuronal activity changes in the prefrontal cortex (PFC), basal lateral amygdala (BLA), and ventral tegmental area (VTA). Chemogenetic manipulation, optogenetic recording, and in vivo calcium imaging identify that the PFC to BLA pathway is causally linked to heightened aggression in stressed males, and the PFC to VTA pathway is causally linked to social withdrawal in stressed females. Isolation stress induces genome-wide transcriptional alterations in a region-specific manner. Particularly, the upregulated genes in BLA of stressed males are under the control of activated transcription factor CREB, and CREB inhibition in BLA normalizes gene expression and reverses aggressive behaviors. On the other hand, neuropeptide Hcrt (Hypocretin/Orexin) is among the top-ranking downregulated genes in VTA of stressed females, and Orexin-A treatment rescues social withdrawal. These results have revealed molecular mechanisms and potential therapeutic targets for stress-related mental illness.

CO2 signaling mediates neurovascular coupling in the cerebral cortex

Neurovascular coupling is a fundamental brain mechanism that regulates local cerebral blood flow (CBF) in response to changes in neuronal activity. Functional imaging techniques are commonly used to record these changes in CBF as a proxy of neuronal activity to study the human brain. However, the mechanisms of neurovascular coupling remain incompletely understood. Here we show in experimental animal models (laboratory rats and mice) that the neuronal activity-dependent increases in local CBF in the somatosensory cortex are prevented by saturation of the CO2-sensitive vasodilatory brain mechanism with surplus of exogenous CO2 or disruption of brain CO2/HCO3− transport by genetic knockdown of electrogenic sodium-bicarbonate cotransporter 1 (NBCe1) expression in astrocytes. A systematic review of the literature data shows that CO2 and increased neuronal activity recruit the same vasodilatory signaling pathways. These results and analysis suggest that CO2 mediates signaling between neurons and the cerebral vasculature to regulate brain blood flow in accord with changes in the neuronal activity.

Leptin in the Commissural Nucleus of the Tractus Solitarius (cNTS) and Anoxic Stimulus in the Carotid Body Chemoreceptors Increases cNTS Leptin Signaling Receptor and Brain Glucose Retention in Rats.

Background and Objectives: The commissural nucleus of the tractus solitarius (cNTS) not only responds to glucose levels directly, but also receives afferent signals from the liver, and from the carotid chemoreceptors (CChR). In addition, leptin, through its receptors in the cNTS, regulates food intake, body weight, blood glucose levels, and brain glucose retention (BGR). These leptin effects on cNTS are thought to be mediated through the sympathetic–adrenal system. How these different sources of information converging in the NTS regulate blood glucose levels and brain glucose retention remains largely unknown. The goal of the present study was to determine whether the local administration of leptin in cNTS alone, or after local anoxic stimulation using sodium cyanide (NaCN) in the carotid sinus, modifies the expression of leptin Ob-Rb and of c-Fos mRNA. We also investigated how leptin, alone, or in combination with carotid sinus stimulation, affected brain glucose retention. Materials and Methods: The experiments were carried out in anesthetized male Wistar rats artificially ventilated to maintain homeostatic values for pO2, pCO2, and pH. We had four groups: (a) experimental 1, leptin infusion in cNTS and NaCN in the isolated carotid sinus (ICS; n = 10); (b) experimental 2, leptin infusion in cNTS and saline in the ICS (n = 10); (c) control 1, artificial cerebrospinal fluid (aCSF) in cNTS and NaCN in the ICS (n = 10); (d) control 2, aCSF in cNTS and saline in the ICS (n = 10). Results: Leptin in cNTS, preceded by NaCN in the ICS increased BGR and leptin Ob-Rb mRNA receptor expression, with no significant increases in c-Fos mRNA in the NTSc. Conclusions: Leptin in the cNTS enhances brain glucose retention induced by an anoxic stimulus in the carotid chemoreceptors, through an increase in Ob-Rb receptors, without persistent changes in neuronal activation.

Features of Brain Rhythms During the Visual Memorizing Task and Auditory Stimuli Using Electroencephalography

Introduction: Brain rhythms are usually measured using electroencephalography (EEG) to detect changes in neuronal activity associated with specific human activity and behavior. This research determines the effectiveness of auditory stimulation for visual memory based on the features of brain rhythm. Methods: The research design involved three main stages: data acquisition, data processing, and data analysis. Sixty university students were selected as participants. They are inquired to memorize and recalled the visual memory assessment under exposure to Mozart’s Sonata music and white noise stimulation with EEG recording. The data acquisition was obtained using 10-20 electrode placement of EEG imaging technique. The wavelet transform approach was used to process the EEG dataset, and the features were statistically analyzed using a t-test. Results: Participants were found to memorize better under the white noise stimulation with a mean score of 64.0%. Based on statistical analysis, it was shown that the stimuli had a significant impact on the EEG voltage trend and relative rhythm power. The relative alpha, gamma, and beta power were higher when exposed to white noise, indicating that alertness and sensory information processing were improved. Conclusion: Therefore, it showed that memorizing visual memory tasks under white noise auditory stimulation had improved participants memory due to activation of certain brain rhythms at specific locations.

Activation of LH GABAergic inputs counteracts fasting-induced changes in tVTA/RMTG neurons.

Dopamine neurons in the ventral tegmental area (VTA) are strongly innervated by GABAergic neurons in the 'tail of the VTA' (tVTA), also known as the rostralmedial tegmental nucleus (RMTg). Disinhibition of dopamine neurons through firing of the GABAergic neurons projecting from the lateral hypothalamus (LH) leads to reward seeking and consumption through dopamine release in the nucleus accumbens. VTA dopamine neurons respond to changes in motivational state, yet less is known about whether tVTA/RMTg GABAergic neurons or the LH GABAergic neurons that project to them are also affected by changes in motivational state, such as fasting. An acute 16h overnight fast decreased the excitability of tVTA/RMTg GABAergic neurons of male and female mice. In addition, fasting decreased synaptic strength at LH GABA to tVTA/RMTg GABAergic synapses, indicated by reduced amplitude of optically evoked currents, decreased readily releasable pool (RRP) size and replenishment. Optical stimulation of LH GABA terminals suppressed evoked action potentials of tVTA/RMTg GABAergic neurons in unfasted mice, but this effect decreased following fasting. Furthermore, during fasting, LH GABA inputs to tVTA/RMTg neurons maintained functional connectivity during depolarization, as depolarization block was reduced following fasting. Taken together, inhibitory synaptic transmission from LH GABA inputs onto tVTA/RMTg GABAergic neurons decreases following fasting; however, ability to functionally inhibit tVTA/RMTg GABAergic neurons is preserved, allowing for possible disinhibition of dopamine neurons and subsequent foraging. KEY POINTS: While dopamine neuronal activity changes with motivational state, it is unknown if fasting influences tVTA/RMTg GABAergic neurons, a major inhibitory input to ventral tegmental area (VTA) dopamine neurons. In unfasted mice, there were sex differences in inhibitory synaptic transmission onto tVTA/RMTg GABAergic neurons. Activation of lateral hypothalamus (LH) GABAergic neurons decreases firing of tVTA/RMTg GABAergic neurons through a monosynaptic input. An acute fast decreases the excitability of tVTA/RMTg GABAergic neurons. An acute fast decreases inhibitory synaptic transmission of the LH GABA input to tVTA/RMTg GABAergic neurons in both male and female mice.

An increase in spontaneous activity mediates visual habituation.

SUMMARYThe cerebral cortex is spontaneously active, but the function of this ongoing activity remains unclear. To test whether spontaneous activity encodes learned experiences, we measured the response of neuronal populations in mouse primary visual cortex with chronic two-photon calcium imaging during visual habituation to a specific oriented stimulus. We find that, during habituation, spontaneous activity increases in neurons across the full range of orientation selectivity, eventually matching that of evoked levels. This increase in spontaneous activity robustly correlates with the degree of habituation. Moreover, boosting spontaneous activity with two-photon optogenetic stimulation to the levels of visually evoked activity accelerates habituation. Our study shows that cortical spontaneous activity is linked to habituation, and we propose that habituation unfolds by minimizing the difference between spontaneous and stimulus-evoked activity levels. We conclude that baseline spontaneous activity could gate incoming sensory information to the cortex based on the learned experience of the animal.

DNA damage and repair in the visual center in the rhesus monkey model of glaucoma

To study the DNA damage and repair methods of visual central neurons in a glaucoma model, a rhesus monkey chronic glaucoma model was established by laser induction, and changes in intraocular pressure (IOP), the optic cup fundus, the thickness of the retinal nerve fiber layer and the diameter of the optic nerve were evaluated. After a sufficient period of time, the model was euthanized, and the lateral geniculate body, primary visual cortex (V1 region) and secondary visual cortex (V2 region) were removed. Through immunofluorescence, ELISA and western blotting assays, the expressions of 8-hydroxyguanosine (8-OHG), a biomarker of oxidative stress, and γH2AX, a marker of DNA double-strand breaks, in the neurons of the LGN, V1 and V2 in the glaucoma model were higher than those of the control group (P < 0.05). The expression of key DNA repair proteins Ku80, Mre11, PCNA, DNA ligase IV and APE1 antibodies in the LGN, V1 and V2 of the glaucoma model was higher than that of the control group (P < 0.05), and in the positive TUNEL cells, the levels of cleaved caspase 3, Beclin 1 and LC3B-II/LC3B-I were significantly increased in the LGN of the glaucoma model (P < 0.05), but there was no significant positive expression in the V1 and V2 regions of the glaucoma model compared with the normal control group (P > 0.05). Transmission electron microscopy also showed that apoptotic bodies and autolysosomes (changes in neuronal apoptosis and autophagy activation) appeared in some neurons of the LGN in glaucoma, but there were no significant abnormal changes in the V1 and V2 regions of glaucoma or in any specimens in the normal group. In terms of neuron counting, the number of neurons in the LGN of the glaucoma model was lower than that in the normal control group (P < 0.05), but there was no significant difference in the number of neurons in the V1 and V2 regions between the two groups (P > 0.05). Similarly, the expression of glial cells in the LGN, V1 and V2 of the glaucoma model was higher than that in the control group (P < 0.05). Therefore, the results showed that DNA oxidative damage and various repair processes occurred in neurons of the LGN, V1 and V2 of the glaucoma model, and finally, LGN neurons died in the glaucoma model.

Deep brain stimulation of the anterior nuclei of the thalamus can alleviate seizure severity and induce hippocampal GABAergic neuronal changes in a pilocarpine-induced epileptic mouse brain.

Deep brain stimulation (DBS) of the anterior nucleus of the thalamus (ANT) has been widely used as an effective treatment for refractory temporal lobe epilepsy. Despite its promising clinical outcome, the exact mechanism of how ANT-DBS alleviates seizure severity has not been fully understood, especially at the cellular level. To assess effects of DBS, the present study examined electroencephalography (EEG) signals and locomotor behavior changes and conducted immunohistochemical analyses to examine changes in neuronal activity, number of neurons, and neurogenesis of inhibitory neurons in different hippocampal subregions. ANT-DBS alleviated seizure activity, abnormal locomotor behaviors, reduced theta-band, increased gamma-band EEG power in the interictal state, and increased the number of neurons in the dentate gyrus (DG). The number of parvalbumin- and somatostatin-expressing inhibitory neurons was recovered to the level in DG and CA1 of naïve mice. Notably, BrdU-positive inhibitory neurons were increased. In conclusion, ANT-DBS not only could reduce the number of seizures, but also could induce neuronal changes in the hippocampus, which is a key region involved in chronic epileptogenesis. Importantly, our results suggest that ANT-DBS may lead to hippocampal subregion-specific cellular recovery of GABAergic inhibitory neurons.

Environmental enrichment recruits activin A to recalibrate neural activity in mouse hippocampus.

The TGF-β family member activin A modulates neural underpinnings of cognitive and affective functions in an activity-dependent fashion. We have previously shown that exploration of a novel and enriched environment (EE) strongly enhanced activin signaling. Whereas the many beneficial effects of EE are amply documented, the underlying mechanisms remain largely elusive. Here, we examined the hypothesis that EE recruits activin to regulate synaptic plasticity in a coordinated, cognition-promoting manner. Elevated activin levels after EE enhanced CA1 pyramidal cell excitability, facilitated synaptic transmission, and promoted long-term potentiation. These EE-induced changes were largely absent in mice expressing a dominant-negative mutant of activin receptor IB. We then interrogated the impact of activin on network oscillations and functional connectivity, using high-speed Ca 2+ imaging to study spike routing within networks formed by dissociated primary hippocampal cultures. Activin facilitated Ca2+ signaling, enhanced the network strength, and shortened the weighted characteristic path length. In the slice preparation, activin promoted theta oscillations during cholinergic stimulation. Thus, we advance activin as an activity-dependent and very early molecular effector that translates behavioral stimuli experienced during EE exposure into a set of synchronized changes in neuronal excitability, synaptic plasticity, and network activity that are all tuned to improve cognitive functions.

Differential Activation of Pain Circuitry Neuron Populations in a Mouse Model of Spinal Cord Injury-Induced Neuropathic Pain

Neuropathic pain (NP) is one of the most common and debilitating comorbidities of spinal cord injury (SCI). Current therapies are often ineffective due in part to an incomplete understanding of underlying pathogenic mechanisms. In particular, it remains unclear how SCI leads to dysfunction in the excitability of nociceptive circuitry. The immediate early gene c-Fos has long been used in pain processing locations as a marker of neuronal activation. We employed a mouse reporter line with fos-promoter driven Cre-recombinase to define neuronal activity changes in relevant pain circuitry locations following cervical spinal cord level (C)5/6 contusion (using both females and males), a SCI model that results in multiple forms of persistent NP-related behavior. SCI significantly increased activation of cervical dorsal horn (DH) projection neurons, as well as induced a selective reduction in the activation of a specific DH projection neuron subpopulation that innervates the periaqueductal gray (PAG), an important brain region involved in descending inhibitory modulation of DH pain transmission. SCI also increased the activation of both protein kinase C (PKC)γ and calretinin excitatory DH interneuron populations. Interestingly, SCI promoted a significant decrease in the activation selectively of neuronal nitric oxide synthase (nNOS)-expressing inhibitory interneurons of cervical DH. In addition, SCI altered activation of various supraspinal neuron populations associated with pain processing, including a large increase in thalamus and a significant decrease in PAG. These findings reveal a complex and diverse set of SCI-induced neuron activity changes across the pain circuitry neuraxis. Moving forward, these results can be used to inform therapeutic targeting of defined neuronal populations in NP.SIGNIFICANCE STATEMENT Neuropathic pain (NP) is one of the most common and highly debilitating comorbidities of spinal cord injury (SCI). Unfortunately, current therapies are often ineffective due in part to an incomplete understanding of underlying pathogenic mechanisms. In particular, it remains unclear how SCI leads to dysfunction in excitability of nociceptive circuitry. Using a FosTRAP2 reporter mouse line in a model of SCI-induced NP, we show SCI alters activation of a number of important interneuron and projection neuron populations across relevant spinal cord and brain locations of the pain circuitry neuraxis. These data suggest a role for maladaptive plasticity involving specific subpopulations of neurons and circuits in driving SCI-induced chronic pain. Moving forward, these results can be used to inform therapeutic targeting of defined neuronal populations in NP.

Changes in the excitability of primary hippocampal neurons following exposure to 3.0 GHz radiofrequency electromagnetic fields

Exposures to radiofrequency electromagnetic fields (RF-EMFs, 100 kHz to 6 GHz) have been associated with both positive and negative effects on cognitive behavior. To elucidate the mechanism of RF-EMF interaction, a few studies have examined its impact on neuronal activity and synaptic plasticity. However, there is still a need for additional basic research that further our understanding of the underlying mechanisms of RF-EMFs on the neuronal system. The present study investigated changes in neuronal activity and synaptic transmission following a 60-min exposure to 3.0 GHz RF-EMF at a low dose (specific absorption rate (SAR) < 1 W/kg). We showed that RF-EMF exposure decreased the amplitude of action potential (AP), depolarized neuronal resting membrane potential (MP), and increased neuronal excitability and synaptic transmission in cultured primary hippocampal neurons (PHNs). The results show that RF-EMF exposure can alter neuronal activity and highlight that more investigations should be performed to fully explore the RF-EMF effects and mechanisms.

The Origin of Abnormal Beta Oscillations in the Parkinsonian Corticobasal Ganglia Circuits.

Parkinson's disease (PD) is a neurodegenerative brain disorder associated with motor and nonmotor symptoms. Exaggerated beta band (15–30 Hz) neuronal oscillations are widely observed in corticobasal ganglia (BG) circuits during parkinsonism. Abnormal beta oscillations have been linked to motor symptoms of PD, but their exact relationship is poorly understood. Nevertheless, reduction of beta oscillations can induce therapeutic effects in PD patients. While it is widely believed that the external globus pallidus (GPe) and subthalamic nucleus (STN) are jointly responsible for abnormal rhythmogenesis in the parkinsonian BG, the role of other cortico-BG circuits cannot be ignored. To shed light on the origin of abnormal beta oscillations in PD, here we review changes of neuronal activity observed in experimental PD models and discuss how the cortex and different BG nuclei cooperate to generate and stabilize abnormal beta oscillations during parkinsonism. This may provide further insights into the complex relationship between abnormal beta oscillations and motor dysfunction in PD, which is crucial for potential target-specific therapeutic interventions in PD patients.

Morpho-Electric Properties and Diversity of Oxytocin Neurons in Paraventricular Nucleus of Hypothalamus in Female and Male Mice.

Oxytocin (OXT) neurons in paraventricular nucleus of hypothalamus (PVN) are involved in modulating multiple functions, including social, maternal, feeding, and emotional related behaviors. PVN OXT neurons are canonically classified into magnocellular (Magno) and parvocellular (Parvo) subtypes. However, morpho-electric properties and the diversity of PVN OXT neurons are not well investigated. In this study, we profiled the morpho-electric properties of PVN OXT neurons by combining transgenic mice, electrophysiological recording, morphologic reconstruction, and unsupervised clustering analyses. Total 224 PVN OXT neurons from 23 mice were recorded and used for analyses in this study, and 29 morpho-electric parameters were measured. Magno and Parvo OXT neurons have prominent differences in their morpho-electric features, and PVN OXT neurons in male and female mice share similar neuronal properties. Some morpho-electric features of PVN OXT neurons, especially Magno neurons, exhibit significant diverse changes along the rostral-caudal axis. Furthermore, we find that PVN OXT neurons are classified into at least six subtypes based on their morpho-electric properties via unsupervised clustering. Only one Magno-Parvo mixed subtype in posterior PVN subregion, but not the other five subtypes, showed significant neuronal activity change in different feeding conditions. Our study supports the diversity of PVN OXT neurons and subtle neuron classification will promote excavating the functions of oxytocinergic system.SIGNIFICANCE STATEMENT Oxytocin (OXT) is well known for its function in labor induction, but it also plays multiple roles in social, feeding, and emotional behaviors via modulating different brain regions. Paraventricular nucleus of hypothalamus (PVN) OXT neurons are traditionally classified into magnocellular and parvocellular. However, functional and single-cell transcriptomic studies indicate that OXT neurons should be further classified. Here, we thoroughly investigated the morpho-electric properties and spatial distribution of PVN OXT neurons, and find that OXT neurons have at least six subtypes based on their morpho-electric features. Among these six subtypes, only one magnocellular-parvocellular mixed subtype, which are distributed in the posterior PVN subregion, change their activities with different feeding states. Our study uncovers the diversity of PVN OXT neurons and suggests the necessary of subtle neuronal classification.

Identification of Brain Regions Activated by Sevoflurane and Propofol and Regional Changes in Gene Expression.

General anesthetics have different efficacies and side effect incidences based on their mechanism of action. However, detailed comparative studies of anesthetics are incomplete. In this study, target brain regions and gene expression changes in these brain regions were determined for sevoflurane and propofol to understand the mechanisms that cause differences among anesthetics. Rats were anesthetized with sevoflurane or propofol for 1 hr, and brain regions with anesthesia-induced changes in neuronal activity were examined by immunohistochemistry and in situ hybridization for c-Fos. Among the identified target brain regions, gene expression analysis was performed in the habenula, the solitary nucleus and the medial vestibular nucleus from laser microdissected samples. Genes altered by sevoflurane and propofol were different and included genes involved in the incidence of postoperative nausea and vomiting and emergence agitation, such as Egr1 and Gad2. GO enrichment analysis showed that the altered genes tended to be evenly distributed in all functional category. The detailed profiles of target brain regions and induced gene expression changes of sevoflurane and propofol in this study will provide a basis for analyzing the effects of each anesthetic agent and the risk of adverse events.