Reconstructing rodent brain signals during euthanasia with eigensystem realization algorithm (ERA).

We accurately reconstruct the Local Field Potential time series obtained from anesthetized and awake rats, both before and during CO 2 euthanasia. We apply the Eigensystem Realization Algorithm to identify an underlying linear dynamical system capable of generating the observed data. Time series exhibiting more intricate dynamics typically lead to systems of higher dimensions, offering a means to assess the complexity of the brain throughout various phases of the experiment. Our results indicate that anesthetized brains possess complexity levels similar to awake brains before CO 2 administration. This resemblance undergoes significant changes following euthanization, as signals from the awake brain display a more resilient complexity profile, implying a state of heightened neuronal activity or a last fight response during the euthanasia process. In contrast, anesthetized brains seem to enter a more subdued state early on. Our data-driven techniques can likely be applied to a broader range of electrophysiological recording modalities.

The Fibromyalgia Pain Experience: A Scoping Review of the Preclinical Evidence for Replication and Treatment of the Affective and Cognitive Pain Dimensions.

Fibromyalgia is a chronic, widespread pain disorder that is strongly represented across the affective and cognitive dimensions of pain, given that the underlying pathophysiology of the disorder is yet to be identified. These affective and cognitive deficits are crucial to understanding and treating the fibromyalgia pain experience as a whole but replicating this multidimensionality on a preclinical level is challenging. To understand the underlying mechanisms, animal models are used. In this scoping review, we evaluate the current primary animal models of fibromyalgia regarding their translational relevance within the affective and cognitive pain realms, as well as summarize treatments that have been identified preclinically for attenuating these deficits.

Multi-region local field potential signatures and brain coherence alternations in response to nitroglycerin-induced migraine attacks.

OBJECTIVE: To decipher the underlying mechanisms of nitroglycerin (NTG)-induced migraine electrophysiologically. BACKGROUND: Migraine is a recurrent primary headache disorder with moderate to severe disability; however, the pathophysiology is not fully understood. Consequently, safe and effective therapies to alleviate migraine headaches are limited. Local field potential (LFP) recording, as a neurophysiological tool, has been widely utilized to investigate combined neuronal activity. METHODS: We recorded LFP changes simultaneously from the anterior cingulate cortex, posterior nucleus of the thalamus, trigeminal ganglion, and primary visual cortex after NTG injection in both anesthetized and freely moving rats. Additionally, brain coherence was processed, and light-aversive behavior measurements were implemented. RESULTS: Significant elevations of LFP powers with various response patterns for the delta, theta, alpha, beta, and gamma bands following NTG injection were detected in both anesthetized and freely moving rats; however, a surge of coherence alternations was exclusively observed in freely moving rats after NTG injection. CONCLUSION: The multi-region LFP signatures and brain coherence alternations in response to NTG-induced migraine attacks were determined. Furthermore, the results of behavior measurements in the freely moving group indicated that NTG induced the phenomenon of photophobia in our study. All these findings offer novel insights into the interpretation of migraine mechanisms and related treatments.

Multi-region local field potential signatures in response to the formalin-induced inflammatory stimulus in male rats.

Pain can be ignited by noxious chemical (e.g., acid), mechanical (e.g., pressure), and thermal (e.g., heat) stimuli and generated by the activation of sensory neurons and their axonal terminals called nociceptors in the periphery. Nociceptive information transmitted from the periphery is projected to the central nervous system (thalamus, somatosensory cortex, insular, anterior cingulate cortex, amygdala, periaqueductal grey, prefrontal cortex, etc.) to generate a unified experience of pain. Local field potential (LFP) recording is one of the neurophysiological tools to investigate the combined neuronal activity, ranging from several hundred micrometers to a few millimeters (radius), located around the embedded electrode. The advantage of recording LFP is that it provides stable simultaneous activities in various brain regions in response to external stimuli. In this study, differential LFP activities from the contralateral anterior cingulate cortex (ACC), ventral tegmental area (VTA), and bilateral amygdala in response to peripheral noxious formalin injection were recorded in anesthetized male rats. The results indicated increased power of delta, theta, alpha, beta, and gamma bands in the ACC and amygdala but no change of gamma-band in the right amygdala. Within the VTA, intensities of the delta, theta, and beta bands were only enhanced significantly after formalin injection. It was found that the connectivity (i.t. the coherence) among these brain regions reduced significantly under the formalin-induced nociception, which suggests a significant interruption within the brain. With further study, it will sort out the key combination of structures that will serve as the signature for pain state.

Evidence and explanation for the involvement of the nucleus accumbens in pain processing.

The nucleus accumbens (NAc) is a subcortical brain structure known primarily for its roles in pleasure, reward, and addiction. Despite less focus on the NAc in pain research, it also plays a large role in the mediation of pain and is effective as a source of analgesia. Evidence for this involvement lies in the NAc's cortical connections, functions, pharmacology, and therapeutic targeting. The NAc projects to and receives information from notable pain structures, such as the prefrontal cortex, anterior cingulate cortex, periaqueductal gray, habenula, thalamus, etc. Additionally, the NAc and other pain-modulating structures share functions involving opioid regulation and motivational and emotional processing, which each work beyond simply the rewarding experience of pain offset. Pharmacologically speaking, the NAc responds heavily to painful stimuli, due to its high density of µ opioid receptors and the activation of several different neurotransmitter systems in the NAc, such as opioids, dopamine, calcitonin gene-related peptide, γ-aminobutyric acid, glutamate, and substance P, each of which have been shown to elicit analgesic effects. In both preclinical and clinical models, deep brain stimulation of the NAc has elicited successful analgesia. The multi-functional NAc is important in motivational behavior, and the motivation for avoiding pain is just as important to survival as the motivation for seeking pleasure. It is possible, then, that the NAc must be involved in both pleasure and pain in order to help determine the motivational salience of positive and negative events.

A model of pain behaviors in freely moving rats generated by controllable electrical stimulation of the peripheral nerve.

BACKGROUND: Neuropathic pain patients have described experiencing unprovoked, intermittent pain attacks with shooting, stabbing, and burning qualities. Rodent models used in previous literature usually only involve acute exposure, and/or are unable to manipulate the stimulation intensity in vivo by the experimenter during an experiment. NEW METHOD: This paper describes a method to induce controllable pain behaviors in rodents using a wireless portable electronic device that can be manipulated within the course of an experiment. A stimulating electrode was implanted at the L5 spinal nerve location in Sprague-Dawley rats and our custom-built wireless stimulating device was attached to deliver variable stimulation in freely moving animals (50 Hz, 0.5 V; 100 Hz, 1 V). RESULTS: Implantation itself did not induce hypersensitivity as measured by the mechanical paw withdrawal threshold test. Observation of pain behaviors (paw elevation and licking) indicated that high stimulation intensity yielded a significant increase in pain behaviors. Even further, high intensity stimulation resulted in a behavioral "wind-up" of pain behaviors that persisted into the resting period when no stimulation was applied. COMPARISON WITH EXISTING METHODS AND CONCLUSIONS: This method can be used to study pain behaviors in a controllable way in freely moving rodents in comparison to existing models that are acute and/or are unable to manipulate the stimulation intensity in vivo.

Role for the Ventral Posterior Medial/Posterior Lateral Thalamus and Anterior Cingulate Cortex in Affective/Motivation Pain Induced by Varicella Zoster Virus.

Varicella zoster virus (VZV) infects the face and can result in chronic, debilitating pain. The mechanism for this pain is unknown and current treatment is often not effective, thus investigations into the pain pathway become vital. Pain itself is multidimensional, consisting of sensory and affective experiences. One of the primary brain substrates for transmitting sensory signals in the face is the ventral posterior medial/posterior lateral thalamus (VPM/VPL). In addition, the anterior cingulate cortex (ACC) has been shown to be vital in the affective experience of pain, so investigating both of these areas in freely behaving animals was completed to address the role of the brain in VZV-induced pain. Our lab has developed a place escape avoidance paradigm (PEAP) to measure VZV-induced affective pain in the orofacial region of the rat. Using this assay as a measure of the affective pain experience a significant response was observed after VZV injection into the whisker pad and after VZV infusion into the trigeminal ganglion. Local field potentials (LFPs) are the summed electrical current from a group of neurons. LFP in both the VPM/VPL and ACC was attenuated in VZV injected rats after inhibition of neuronal activity. This inhibition of VPM/VPL neurons was accomplished using a designer receptor exclusively activated by a designer drug (DREADD). Immunostaining showed that cells within the VPM/VPL expressed thalamic glutamatergic vesicle transporter-2, NeuN and DREADD suggesting inhibition occurred primarily in excitable neurons. From these results we conclude: (1) that VZV associated pain does not involve a mechanism exclusive to the peripheral nerve terminals, and (2) can be controlled, in part, by excitatory neurons within the VPM/VPL that potentially modulate the affective experience by altering activity in the ACC.

Inflammatory pain by carrageenan recruits low-frequency local field potential changes in the anterior cingulate cortex.

The anterior cingulate cortex (ACC) has been extensively cited as a key area for processing pain affect. While local field potential (LFP) studies in other fields have yielded a great deal of information about neural oscillations, there is a poverty in the pain literature about the neural LFP profile related to pain, particularly in freely moving animals. In this study, we revealed the LFP profile in the ACC in freely moving rats during carrageenan inflammation. Mechanical allodynia was recorded before and after unilateral injection of carrageenan/saline in the left hindpaw. LFP activity in the ACC was recorded at baseline, after injection, and after injection with mechanical stimulation to the paw using a von Frey filament. This study uniquely reveals that carrageenan injection significantly recruited ACC LFP activity in delta, theta, and alpha bands (0-13Hz). Application of von Frey mechanical stimulation to the carrageenan-injected paw resulted in a significant increase in delta, theta, and alpha bands over and above what was recruited by carrageenan alone and further expanded the LFP range to additionally include beta activity (13-30Hz). Taken together, these data reveal significant changes in the lowest-frequency activities in the LFP range during painful inflammation, which merit attention. LFP is a powerful window to reveal wide-range, integrated synaptic processing by low-frequency cellular events during behavior. Information about LFP during pain broadens the scope of our understanding of pain mechanisms, our greatest resource for designing management approaches.

Local field potentials in the ventral tegmental area during cocaine-induced locomotor activation: Measurements in freely moving rats.

The ventral tegmental area (VTA) has been established as a critical nucleus for processing behavioral changes that occur during psychostimulant use. Although it is known that cocaine induced locomotor activity is initiated in the VTA, not much is known about the electrical activity in real time. The use of our custom-designed wireless module for recording local field potential (LFP) activity provides an opportunity to confirm and identify changes in neuronal activity within the VTA of freely moving rats. The purpose of this study was to investigate the changes in VTA LFP activity in real time that underlie cocaine induced changes in locomotor behavior. Recording electrodes were implanted in the VTA of rats. Locomotor behavior and LFP activity were simultaneously recorded at baseline, and after saline and cocaine injections. Results indicate that cocaine treatment caused increases in both locomotor behavior and LFP activity in the VTA. Specifically, LFP activity was highest during the first 30 min following the cocaine injection and was most robust in Delta and Theta frequency bands; indicating the role of low frequency VTA activity in the initiation of acute stimulant-induced locomotor behavior. Our results suggest that LFP recording in freely moving animals can be used in the future to provide valuable information pertaining to drug induced changes in neural activity.

A digital wireless system for closed-loop inhibition of nociceptive signals.

Neurostimulation of the spinal cord or brain has been used to inhibit nociceptive signals in pain management applications. Nevertheless, most of the current neurostimulation models are based on open-loop system designs. There is a lack of closed-loop systems for neurostimulation in research with small freely-moving animals and in future clinical applications. Based on our previously developed analog wireless system for closed-loop neurostimulation, a digital wireless system with real-time feedback between recorder and stimulator modules has been developed to achieve multi-channel communication. The wireless system includes a wearable recording module, a wearable stimulation module and a transceiver connected to a computer for real-time and off-line data processing, display and storage. To validate our system, wide dynamic range neurons in the spinal cord dorsal horn have been recorded from anesthetized rats in response to graded mechanical stimuli (brush, pressure and pinch) applied in the hind paw. The identified nociceptive signals were used to automatically trigger electrical stimulation at the periaqueductal gray in real time to inhibit their own activities by the closed-loop design. Our digital wireless closed-loop system has provided a simplified and efficient method for further study of pain processing in freely-moving animals and potential clinical application in patients.

A closed loop feedback system for automatic detection and inhibition of mechano-nociceptive neural activity.

Clinical studies have shown that spinal or cerebral neurostimulation can significantly relieve pain. Current neurostimulators work in an open loop; hence, their efficacy depends on the patient's or physician's comprehension of pain. We have proposed and developed a real-time automatic recognition program with signal processing functions to detect action potentials. By using a wireless neurorecording module, spinal neuronal responses to mechanical stimuli (brush, pressure, and pinch) applied to rats' hind paws were recorded. Nociceptive spinal responses were detected and suppressed by our automated module through delivering electrical stimulation to the periaqueductal gray (PAG). The interspike intervals (ISIs) of the fired action potentials were used to distinguish among the three different mechanical stimuli. Our system was able to detect the neuronal activity intensities and deliver trigger signals to the neurostimulator according to a pre-set threshold in a closed-loop feedback configuration, thereby suppressing excessive activity in spinal cord dorsal horn neurons.

Detection of thermal pain in rodents through wireless electrocorticography.

In an effort to detect pain in an objective way, Electrocorticography (ECoG) signals were acquired from male Sprague-Dawley rats in response to thermally induced pain. A wearable, wireless multichannel system was utilized to acquire signals from freely-behaving animals during the experiments. ECoG signals were recorded before (baseline) and during the heat exposure for which animals withdrew their paws in response to the painful feeling. Analysis of the signals revealed a clear, high-amplitude peak at the moment of the paw withdrawal across all four recording channels in each test. Analysis in the frequency domain found the peaks coincided with an abrupt increase of delta rhythms (under 4 Hz). In the baseline, heating, and post-withdrawal segments, these rhythms were relatively low, indicating that the sharp increase in delta activity might be associated with pain. Theta, alpha, beta, and gamma rhythms were also measured, but no significant differences were found between each phase of the signals. These preliminary results are promising; however, more animal models will need to be tested to provide statistically significant results with high confidence.

Inhibition of spinal cord dorsal horn neuronal activity by electrical stimulation of the cerebellar cortex.

The cerebellum plays a major role in not only modulating motor activity, but also contributing to other functions, including nociception. The intermediate hemisphere of the cerebellum receives sensory input from the limbs. With the extensive connection between the cerebellum to brain-stem structures and cerebral cortex, it is possible that the cerebellum may facilitate the descending system to modulate spinal dorsal horn activity. This study provided the first evidence to support this hypothesis. Thirty-one wide-dynamic-range neurons from the left lumbar and 27 from the right lumbar spinal dorsal horn were recorded in response to graded mechanical stimulation (brush, pressure, and pinch) at the hind paws. Electrical stimulation of the cerebellar cortex of the left intermediate hemisphere significantly reduced spinal cord dorsal horn neuron-evoked responses bilaterally in response to peripheral high-intensity mechanical stimuli. It is concluded that the cerebellum may play a potential antinociceptive role, probably through activating descending inhibitory pathways indirectly.

Differential contribution of electrically evoked dorsal root reflexes to peripheral vasodilatation and plasma extravasation.

BACKGROUND: Dorsal root reflexes (DRRs) are antidromic activities traveling along the primary afferent fibers, which can be generated by peripheral stimulation or central stimulation. DRRs are thought to be involved in the generation of neurogenic inflammation, as indicated by plasma extravasation and vasodilatation. The hypothesis of this study was that electrical stimulation of the central stump of a cut dorsal root would lead to generation of DRRs, resulting in plasma extravasation and vasodilatation. METHODS: Sprague-Dawley rats were prepared to expose spinal cord and L4-L6 dorsal roots under pentobarbital general anesthesia. Electrical stimulation of either intact, proximal or distal, cut dorsal roots was applied while plasma extravasation or blood perfusion of the hindpaw was recorded. RESULTS: While stimulation of the peripheral stump of a dorsal root elicited plasma extravasation, electrical stimulation of the central stump of a cut dorsal root generated significant DRRs, but failed to induce plasma extravasation. However, stimulation of the central stump induced a significant increase in blood perfusion. CONCLUSIONS: It is suggested that DRRs are involved in vasodilatation but not plasma extravasation in neurogenic inflammation in normal animals.

Recognition and inhibition of dorsal horn nociceptive signals within a closed-loop system.

We implemented an integrated system that can acquire neuronal signals from spinal cord dorsal horn neurons, wirelessly transmit the signals to a computer, and recognize the nociceptive signals from three different mechanical stimuli (brush, pressure and pinch). Positive peak detection method was chosen to distinguish between signal spikes. The inter spike intervals (ISIs) were calculated from the identified action potentials (APs) and fed into a numerical array called cluster. When the sum of the ISIs in the cluster reached a critical level, the program recognized the recorded signals as nociceptive inputs. The user has the flexibility to tune both the cluster size and critical threshold for individual's need to reach optimization in pain signal recognition. The program was integrated with a wireless neurostimulator to form a feedback loop to recognize and inhibit nociceptive signals.

Contributions of dorsal root reflex and axonal reflex to formalin-induced inflammation.

The dorsal root reflex (DRR) and the axonal reflex (AR) are antidromic activities in primary afferents and are involved in neurogenic inflammation. DRRs and/or ARs lead to release of neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). CGRP causes blood vessels to dilate leading to an increase in blood perfusion, whereas SP causes plasma extravasation, leading to edema. Both DRR and AR can be evoked by noxious stimuli. The goal of this study was to determine the role of DRR and AR in neurogenic inflammation by examining the blood perfusion (BP) change in hindpaws in response to formalin injection (an acute inflammatory agent). Laser Doppler images were collected simultaneously in both hindpaws in anesthetized rats to determine the level of BP. Local lidocaine was applied to the left sciatic nerve to block both orthodromic signals and antidromic DRRs without affecting ARs. All rats then received a subcutaneous formalin injection to the left hindpaw. Our results showed that (1) the mean BP of the left paw increased significantly following formalin injection, with or without lidocaine; (2) application of lidocaine in the left sciatic nerve alone significantly increased BP ipsilaterally; (3) formalin injection following lidocaine application significantly increased BP more than the group without lidocaine; and (4) there was delayed significant BP increase in the right (contralateral) hindpaw following formalin injection with or without lidocaine. It is concluded that ARs play a more important role than DRRs in formalin-induced neurogenic inflammation.

A combined wireless neural stimulating and recording system for study of pain processing.

Clinical studies have shown that spinal or cortical neurostimulation could significantly improve pain relief. The currently available stimulators, however, are used only to generate specific electrical signals without the knowledge of physiologically responses caused from the stimulation. We thus propose a new system that can adaptively generate the optimized stimulating signals base on the correlated neuron activities. This new method could significantly improve the efficiency of neurostimulation for pain relief. We have developed an integrated wireless recording and stimulating system to transmit the neuronal signals and to activate the stimulator over the wireless link. A wearable prototype has been developed consisting of amplifiers, wireless modules and a microcontroller remotely controlled by a Labview program in a computer to generate desired stimulating pulses. The components were assembled on a board with a size of 2.5 cm x 5 cm to be carried by a rat. To validate our system, lumbar spinal cord dorsal horn neuron activities of anesthetized rats have been recorded in responses to various types of peripheral graded mechanical stimuli. The stimulation at the periaqueductal gray and anterior cingulate cortex with different combinations of electrical parameters showed a comparable inhibition of spinal cord dorsal horns activities in response to the mechanical stimuli. The Labview program was also used to monitor the neuronal activities and automatically activate the stimulator with designated pulses. Our wireless system has provided an opportunity for further study of pain processing with closed-loop stimulation paradigm in a potential new pain relief method.

Electrical stimulation of the primary somatosensory cortex inhibits spinal dorsal horn neuron activity.

Cortical stimulation has been demonstrated to alleviate certain pain conditions. The aim of this study was to determine the responses of the spinal cord dorsal horn neurons to stimulation of the primary somatosensory cortex (SSC). We hypothesized that direct stimulation of the SSC will inhibit the activity of spinal dorsal horn neurons by activating the descending inhibitory system. Thirty-four wide dynamic range spinal dorsal horn neurons were recorded in response to graded mechanical stimulation (brush, pressure, and pinch) at their respective receptive fields while a stepwise electrical stimulation (300 Hz, 0.1 ms, at 10, 20, and 30 V) was applied in the SSC through a bipolar tungsten electrode. The responses to brush at control, 10 V, 20 V, 30 V, and recovery were 16.0 +/- 2.3, 15.8 +/- 2.2, 14.6 +/- 1.8, 14.8 +/- 2.0, and 17.0 +/- 2.2 spikes/s, respectively. The responses to pressure at control, 10 V, 20 V, 30 V, and recovery were 44.7 +/- 5.5, 37.0 +/- 5.6, 29.5 +/- 4.8, 31.6 +/- 5.2, and 43.2 +/- 5.7 spikes/s, respectively. The responses to pinch at control, 10 V, 20 V, 30 V, and recovery were 58.1 +/- 7.0, 42.9 +/- 5.5, 34.8 +/- 3.9, 34.6 +/- 4.4, and 52.6 +/- 6.0 spikes/s, respectively. Significant decreases of the dorsal horn neuronal responses to pressure and pinch were observed during SSC stimulation. It is concluded that electrical stimulation of the SSC produces transient inhibition of the responses of spinal cord dorsal horn neurons to higher intensity mechanical stimuli without affecting innocuous stimuli.

Dorsal horn neuron response patterns to graded heat stimuli in the rat.

Sensory input from various receptors in the periphery first becomes integrated in the spinal cord dorsal horn. The response of the spinal cord dorsal horn neurons to mechanical stimuli are classified as low threshold, high threshold, and wide dynamic range neurons. However, the response pattern of deep dorsal horn cells to heat has not been well described. In this study, the response of the spinal cord dorsal horn neurons to graded heat stimuli were characterized in 147 neurons in rats by extracellular single cell recording. After a differentiable cell was identified, the Peltier heat stimulator was applied to the receptive field and the base temperature was set at 30 degrees C. The heat stimulus was delivered for 10 s from 37-51 degrees C in 2 degrees C increments, with an inter-stimulus interval of 30 s. Out of the 147 neurons, five statistically distinguishable response patterns were identified by latent class cluster analysis. It is concluded that variation of temperature may account for the observed results and indicate functionally different subsets of heat-responsive cells in the deep dorsal horn.

Spinal dorsal horn neuron response to mechanical stimuli is decreased by electrical stimulation of the primary motor cortex.

Motor cortex stimulation (MCS) has been used clinically as a tool for the control for central post-stroke pain and neuropathic facial pain. The underlying mechanisms involved in the antinociceptive effect of MCS are not clearly understood. We hypothesize that the antinociceptive effect is through the modulation of the spinal dorsal horn neuron activity. Thirty-two wide dynamic range spinal dorsal horn neurons were recorded, in response to graded mechanical stimulation (brush, pressure, and pinch) at their respective receptive fields, while a stepwise electrical stimulation was applied simultaneously in the motor cortex. The responses to brush at control, 10 V, 20 V, and 30 V, and recovery were 11.5+/-1.6, 12.1+/-2.6, 11.1+/-2.2, 10.5+/-2.1, and 13.2+/-2.5 spikes/s, respectively. The responses to pressure at control, 10 V, 20 V, and 30 V, and recovery were 33.2+/-6.1, 22.9+/-5.3, 20.5+/-5.0, 17.3+/-3.8, and 27.0+/-4.0 spikes/s, respectively. The responses to pinch at control, 10 V, 20 V, and 30 V, and recovery were 37.2+/-6.4, 26.3+/-4.7, 25.9+/-4.7, 22.5+/-4.3, and 35.0+/-6.2 spikes/s, respectively. It is concluded that, in the rat, electrical stimulation of the motor cortex produces significant transient inhibition of the responses of spinal cord dorsal horn neurons to higher intensity mechanical stimuli without affecting their response to an innocuous stimulus.