open-source-real-time-closed-loop-electrical-threshold-tracking-for

Open-Source Real-Time Closed-Loop Electrical Threshold Tracking for Translational Pain Research

Nociceptors are a class of primary afferent neurons that signal potentially harmful noxious stimuli. An increase in nociceptor excitability occurs in ...

University of Bristol

Rheumatoid arthritis (RA) and rat models of RA exhibit symmetrical mirror-image spread. Many studies have sought to understand the underlying mechanisms and have reported contralateral effects that are manifested in many different forms. It is now well accepted that neurogenic mechanisms contribute to the symmetrical spread of inflammation. However, very few investigators have directly assessed changes in contralateral nerve function and there is a paucity of data. In the present study our aim was to investigate whether there are changes, in particular in the nervous system but also in the vascular system contralateral to an inflamed rat knee joint, that might precede overt inflammation and symmetrical spread. Three to five days following Complete Freund's Adjuvant (CFA) injection we found spontaneous antidromic (away from the CNS) activity in the homologous sensory nerve contralateral to the inflamed joint. Antidromic activity of this nature is known to result in the peripheral release of pro-inflammatory and vasoactive neuropeptides. Importantly, this activity was modulated by systemic analgesic treatment. Furthermore, levels of Evans blue dye extravasation were significantly increased in the joint contralateral to inflammation, indicating altered vascular function. These data suggest that contralateral increases in sensory neural activity and vascular function may account for the symmetrical spread of RA, and that early analgesic treatment may prevent or delay the spread of this debilitating disease.

Sensory nerves have altered function contralateral to a monoarthritis and may contribute to the symmetrical spread of inflammation.

Inflammatory hypersensitivity is characterized by behavioural reductions in withdrawal thresholds to noxious stimuli. Although cutaneous primary afferent neurones are known to have lowered thermal thresholds in inflammation, whether their mechanical thresholds are altered remains controversial. The transient receptor potential channel A1 (TRPA1) is a receptor localized to putative nociceptive neurones and is implicated in mechanical and thermal nociception. Herein, we examined changes in the properties of single primary afferents in normal and acutely inflamed rats and determined whether specific nociceptive properties, particularly mechanical thresholds, are altered in the subpopulation of afferents that responded to the TRPA1 agonist cinnamaldehyde (TRPA1-positive afferents). TRPA1-positive afferents in normal animals belonged to the mechanonociceptive populations, many of which also responded to heat or capsaicin but only a few of which responded to cold. In acute inflammation, a greater proportion of afferents responded to cinnamaldehyde and an increased proportion of dorsal root ganglion neurones expressed TRPA1 protein. Functionally, in inflammation, TRPA1-positive afferents showed significantly reduced mechanical thresholds and enhanced activity to agonist stimulation. Inflammation altered thermal thresholds in both TRPA1-positive and TRPA1-negative afferents. Our data show that a subset of afferents is sensitized to mechanical stimulation by inflammation and that these afferents are defined by expression of TRPA1.

Inflammation reduces mechanical thresholds in a population of transient receptor potential channel A1-expressing nociceptors in the rat.

Thermal sensory testing in rodents informs human pain research. There are important differences in the methodology for delivering thermal stimuli to humans and rodents. This is particularly true in cold pain research. These differences confound extrapolation and de-value nociceptive tests in rodents.

A novel method for delivering ramped cooling reveals rat behaviours at innocuous and noxious temperatures: A comparative study of human psychophysics and rat behaviour.

C-mechanoreceptors in humans comprise a population of unmyelinated afferents exhibiting a wide range of mechanical sensitivities. C-mechanoreceptors are putatively divided into those signaling gentle touch (C-tactile afferents, CTs) and nociception (C-mechanosensitive nociceptors, CMs), giving rise to positive and negative affect, respectively. We sought to distinguish, compare, and contrast the properties of a population of human C-mechanoreceptors to see how fundamental the divisions between these putative subpopulations are. We used microneurography to record from individual afferents in humans and applied electrical and mechanical stimulation to their receptive fields. We show that C-mechanoreceptors can be distinguished unequivocally into two putative populations, comprising CTs and CMs, by electrically evoked spike latency changes (slowing). After both natural mechanical stimulation and repetitive electrical stimulation there was markedly less latency slowing in CTs compared with CMs. Electrical receptive field stimulation, which bypasses the receptor end organ, was most effective in classifying C-mechanoreceptors, as responses to mechanical receptive field stimulation overlapped somewhat, which may lead to misclassification. Furthermore, we report a subclass of low-threshold CM responding to gentle mechanical stimulation and a potential subclass of CT afferent displaying burst firing. We show that substantial differences exist in the mechanisms governing axonal conduction between CTs and CMs. We provide clear electrophysiological "signatures" (extent of latency slowing) that can be used in unequivocally identifying populations of C-mechanoreceptors in single-unit and multiunit microneurography studies and in translational animal research into affective touch. Additionally, these differential mechanisms may be pharmacologically targetable for separate modulation of positive and negative affective touch information. Human skin encodes a plethora of touch interactions, and affective tactile information is primarily signaled by slowly conducting C-mechanoreceptive afferents. We show that electrical stimulation of low-threshold C-tactile afferents produces markedly different patterns of activity compared with high-threshold C-mechanoreceptive nociceptors, although the populations overlap in their responses to mechanical stimulation. This fundamental distinction demonstrates a divergence in affective touch signaling from the first stage of sensory processing, having implications for the processing of interpersonal touch.

Optimal delineation of single C-tactile and C-nociceptive afferents in humans by latency slowing.

Microneurography is the only method for recording from single neurons in intact human nerves. It is challenging - requiring technical expertise, investment in specialised equipment and has sparse data yields.

Ultrasound-guided, open-source microneurography: Approaches to improve recordings from peripheral nerves in man.

Recordings of electrical activity in nerves have provided valuable insights into normal function and pathological behaviours of the nervous system. Current high-resolution techniques (e.g. teased fibre recordings) typically utilise electrodes with a single recording site, capturing the activity of a single isolated neuron per recording.

Multisite silicon probes enable simultaneous recording of spontaneous and evoked activity in multiple isolated C-fibres in rat saphenous nerve.

Pain is a complex polygenic trait whose common genetic underpinnings are relatively ill-defined due in part to challenges in measuring pain as a phenotype. Pain sensitivity can be quantified, but this is difficult to perform at the scale required for genome wide association studies (GWAS). Existing GWAS of pain have identified surprisingly few loci involved in nociceptor function which contrasts strongly with rare monogenic pain states. This suggests a lack of resolution with current techniques. We propose an adaptive methodology within a recall-by-genotype (RbG) framework using detailed phenotyping to screen minor alleles in a candidate 'nociceptor' gene in an attempt to estimate their genetic contribution to pain.

Article	Year
Sensory nerves have altered function contralateral to a monoarthritis and may contribute to the symmetrical spread of inflammation. The European journal of neuroscience\| PubMed ID: 17714187	2007
Inflammation reduces mechanical thresholds in a population of transient receptor potential channel A1-expressing nociceptors in the rat. The European journal of neuroscience\| PubMed ID: 18598259	2008
A novel method for delivering ramped cooling reveals rat behaviours at innocuous and noxious temperatures: A comparative study of human psychophysics and rat behaviour. Journal of neuroscience methods\| PubMed ID: 25857615	2015
Optimal delineation of single C-tactile and C-nociceptive afferents in humans by latency slowing. Journal of neurophysiology\| PubMed ID: 28123010	2017
Ultrasound-guided, open-source microneurography: Approaches to improve recordings from peripheral nerves in man. Clinical neurophysiology : official journal of the International Federation of Clinical Neurophysiology\| PubMed ID: 30107982	2018
Multisite silicon probes enable simultaneous recording of spontaneous and evoked activity in multiple isolated C-fibres in rat saphenous nerve. Journal of neuroscience methods\| PubMed ID: 34800543	2022
Evaluating the association of TRPA1 gene polymorphisms with pain sensitivity: a protocol for an adaptive recall by genotype study. BMC medical genomics\| PubMed ID: 35022050	2022

James P. Dunham

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