<i>In Vivo</i>Calcium Imaging Visualizes Incision-Induced Primary Afferent Sensitization and Its Amelioration by Capsaicin Pretreatment

Ishida, Hirotake; Zhang, Yan; Gomez, Ruben; Shannonhouse, John; Son, Hyeonwi; Banik, Ratan K.; Kim, Yu Shin

doi:10.1523/jneurosci.0457-21.2021

Cited by 11 publications

(8 citation statements)

References 61 publications

(65 reference statements)

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“…So far, few studies have reported on spontaneous activity in sensory neurons using calcium indicators in vivo . In comparison to our baseline data, two studies using a GCaMP3 transgenic mouse line reported much lower levels (0.13-0.19 %) of spontaneously active calcium transients 33,34 . This difference is likely because the overall acquisition rate in these recordings was a lot slower in comparison to ours (<0.2 Hz vs 3.65 Hz).…”

Section: Discussioncontrasting

confidence: 92%

Assessing spontaneous sensory neuron activity usingin vivocalcium imaging

Ingram

Chisholm

Wang

et al. 2023

Preprint

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Heightened spontaneous activity in sensory neurons is often reported in individuals living with chronic pain. It is possible to study this activity in rodents using electrophysiology, but these experiments require great skill and can be prone to bias. Here, we have examined whether in vivo calcium imaging with GCaMP6s can be used as an alternative approach. We show that spontaneously active calcium transients can be visualised in the fourth lumbar dorsal root ganglion (L4 DRG) via in vivo imaging in a mouse model of pain. Application of lidocaine to the nerve, between the inflamed site and the DRG, silenced spontaneous firing and revealed the true baseline level of calcium for spontaneously active neurons. We used this data to train a machine leaning algorithm to predict when a neuron is spontaneously active. We show that our algorithm is accurate in two different models of pain: intraplantar Complete Freund's Adjuvant and antigen-induced arthritis, with accuracies of 90.0% +/- 1.2 and 85.9 % +/-2.1, respectively, assessed against visual inspection by an experienced observer. The algorithm can also detect neuronal activity in imaging experiments generated in a different lab using a different microscope configuration (Accuracy = 94.0 % +/2.2). We provide a Google Colaboratory Notebook to allow anyone easy access to this novel tool, for assessment of peripheral neuron activity in their own calcium imaging setups.

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Section: Discussioncontrasting

confidence: 92%

Assessing spontaneous sensory neuron activity usingin vivocalcium imaging

Ingram

Chisholm

Wang

et al. 2023

Preprint

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“…The plantar incisional surgery was performed as previously reported [ 17 ]. Briefly, mice were anesthetized with sevoflurane delivered via a small nose cone.…”

Section: Methodsmentioning

confidence: 99%

Pretreatment of the ROS Inhibitor Phenyl-N-tert-butylnitrone Alleviates Sleep Deprivation-Induced Hyperalgesia by Suppressing Microglia Activation and NLRP3 Inflammasome Activity in the Spinal Dorsal Cord

et al. 2022

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Sleep deprivation, a common perioperative period health problem, causes ocular discomfort and affects postsurgical pain. However, the mechanism of sleep deprivation-induced increased pain sensitivity is elusive. This study aims to explore the role of ROS in sleep deprivation (SD)-induced hyperalgesia and the underlying mechanism. A 48-h continuous SD was performed prior to the hind paw incision pain modeling in mice. We measured ROS levels, microglial activation, DNA damage and protein levels of iNOS, NLRP3, p-P65 and P65 in mouse spinal dorsal cord. The involvement of ROS in SD-induced prolongation of postsurgical pain was further confirmed by intrathecal injection of ROS inhibitor, phenyl-N-tert-butylnitrone (PBN). Pretreatment of 48-h SD in mice significantly prolonged postsurgical pain recovery, manifesting as lowered paw withdrawal mechanical threshold and paw withdrawal thermal latency. It caused ROS increase and upregulation of iNOS on both Day 1 and 7 in mouse spinal dorsal cord. In addition, upregulation of NLRP3 and p-P65, microglial activation and DNA damage were observed in mice pretreated with 48-h SD prior to the incision. Notably, intrathecal injection of PBN significantly reversed the harmful effects of SD on postsurgical pain recovery, hyperalgesia, microglial activation and DNA damage via the NF-κB signaling pathway. Collectively, ROS increase is responsible for SD-induced hyperalgesia through activating microglial, triggering DNA damage and enhancing NLRP3 inflammasome activity in the spinal dorsal cord.

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“…Indeed, viability of primary DRG neurons as assessed as proportion of high K + responsive neurons did not differ between genotypes. Instead, capsaicin evoked calcium in ux was stronger in DRG neurons of Avil-ChR2 mice suggesting hypersensitivity of transient receptor potential TRPV1 channels, which is a biological correlate of heat pain 71,72 . Calcium imaging results thus agree with the behavioral tests of nociception and reveal the peripheral nociceptive sensitization at a biological level.…”

Section: Discussionmentioning

confidence: 98%

Optogenetic Early Life Pain leads to cortical hyperexcitability, nociceptive hypersensitivity and repetitive behavior

Tegeder

Vogel

Ueberbach

et al. 2022

Preprint

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Pain in early life may affect cortical development and risk of chronic pain. We developed an optogenetic Cre/loxP mouse model of "early-life-pain" (ELP) using mice with transgenic expression of channelrhodopsin-2 (ChR2) under control of the Advillin (Avil) promoter, that drives expression of ChR2 in peripheral somatosensory neurons. Avil-ChR2 (Cre+) and ChR2-flfl control mice were exposed to blue light in a chamber once daily from P1-P5 together with their Cre-negative mother. ELP caused cortical hyperexcitability at P8-9 as assessed via multi-electrode array recordings that coincided with reduced expression of synaptic genes (RNAseq) including Grin2b, neurexins, piccolo and voltage gated calcium and sodium channels which would agree with activity-dependent synaptic pruning. Young adult Avil-ChR2 mice presented with nociceptive hypersensitivity upon heat or mechanical stimulation, which did not resolve up until one year of age. The persistent "pain" phenotype was reflected by capsaicin hypersensitivity in primary sensory neurons of aged mice as assessed by calcium imaging. Avil-ChR2 mice had no disadvantages in Maze tests of anxiety, social interaction and spatial memory, and multiple dimensions of cognitive behavior in IntelliCages. But they stood out by a high rate of repetitive nosepokes, and repetitive corner returns irrespective of success, suggesting inflexibility. Hence, optogenetic early life pain leads to pain in adult mice and repetitive behavior but without cognitive or social deficits.

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In VivoCalcium Imaging Visualizes Incision-Induced Primary Afferent Sensitization and Its Amelioration by Capsaicin Pretreatment

Cited by 11 publications

References 61 publications

Assessing spontaneous sensory neuron activity usingin vivocalcium imaging

Assessing spontaneous sensory neuron activity usingin vivocalcium imaging

Pretreatment of the ROS Inhibitor Phenyl-N-tert-butylnitrone Alleviates Sleep Deprivation-Induced Hyperalgesia by Suppressing Microglia Activation and NLRP3 Inflammasome Activity in the Spinal Dorsal Cord

Optogenetic Early Life Pain leads to cortical hyperexcitability, nociceptive hypersensitivity and repetitive behavior

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