scholarly journals ATP metabolizing enzymes ENPP1, 2 and 3 are localized in sensory neurons of rat dorsal root ganglion

2018 ◽  
Author(s):  
Kentaro Nishida ◽  
Yuka Nomura ◽  
Kanako Kawamori ◽  
Akihiro Ohishi ◽  
Kazuki Nagasawa
Keyword(s):  
Dorsal Root Ganglion ◽  
Sensory Neurons ◽  
Dorsal Root ◽  
Adenine Nucleotide ◽  
Expression Profiles ◽  
Critical Role ◽  
Immunohistochemical Analysis ◽  
Uptake System ◽  
Nucleoside Transporter ◽  
Drg Neurons

In dorsal root ganglion (DRG) neurons, ATP is an important neurotransmitter in nociceptive signaling through P2 receptors (P2Rs) such as P2X2/3R, and adenosine is also involved in anti-nociceptive signaling through adenosine A1R. Thus, the clearance system for adenine nucleotide/nucleoside plays a critical role in regulation of nociceptive signaling, but there is little information on it, especially ectoenzyme expression profiles in DRG. In this study, we examined expression and localization of ecto-nucleotide pyrophosphatase/phosphodiesterases (ENPPs), by which ATP is metabolized to AMP, in rat DRG. The mRNA expression levels of ENPP2 were greater than those of ENPP1 and ENPP3 in rat DRGs. On immunohistochemical analysis, ENPP1, 2 and 3 were found in soma of DRG neurons. Immunopositive rate of ENPP3 was greater than that of ENPP1 and ENPP2 in all DRG neurons. ENPP3, as compared with ENPP1 and ENPP2, was expressed mainly by isolectin B4-positive cells, and slightly by neurofilament 200-positive ones. In this way, the expression profile of ENPP1, 2 and 3 was different in DRGs, and they were mainly expressed in small/medium-sized DRG neurons. Moreover, ENPP1-, 2- and 3-immunoreactivities were colocalized with P2X2R, P2X3R and prostatic acid phosphatase (PAP), as an ectoenzyme for metabolism from AMP to adenosine. Additionally, PAP-immunoreactivity was colocalized with equilibrative nucleoside transporter (ENT) 1, as an adenosine uptake system. These results suggest that the clearance system consisted of ENPPs, PAP and ENT1 plays an important role in regulation of nociceptive signaling in sensory neurons.

Dorsal Root Ganglion Neuron Types and Their Functional Specialization

2018 ◽  
pp. 127-155 ◽  
Author(s):  
Edward C. Emery ◽  
Patrik Ernfors
Keyword(s):  
Chronic Pain ◽  
Dorsal Root Ganglion ◽  
Sensory Neurons ◽  
Dorsal Root ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Drg Neurons ◽  
Low Threshold

Primary sensory neurons of the dorsal root ganglion (DRG) respond and relay sensations that are felt, such as those for touch, pain, temperature, itch, and more. The ability to discriminate between the various types of stimuli is reflected by the existence of specialized DRG neurons tuned to respond to specific stimuli. Because of this, a comprehensive classification of DRG neurons is critical for determining exactly how somatosensation works and for providing insights into cell types involved during chronic pain. This article reviews the recent advances in unbiased classification of molecular types of DRG neurons in the perspective of known functions as well as predicted functions based on gene expression profiles. The data show that sensory neurons are organized in a basal structure of three cold-sensitive neuron types, five mechano-heat sensitive nociceptor types, four A-Low threshold mechanoreceptor types, five itch-mechano-heat–sensitive nociceptor types and a single C–low-threshold mechanoreceptor type with a strong relation between molecular neuron types and functional types. As a general feature, each neuron type displays a unique and predicable response profile; at the same time, most neuron types convey multiple modalities and intensities. Therefore, sensation is likely determined by the summation of ensembles of active primary afferent types. The new classification scheme will be instructive in determining the exact cellular and molecular mechanisms underlying somatosensation, facilitating the development of rational strategies to identify causes for chronic pain.

scholarly journals Cytotoxic Effect of Commercially Available Methylprednisolone Acetate with and without Reduced Preservatives on Dorsal Root Ganglion Sensory Neurons in Rats

2014 ◽  
Vol 5;17 (5;9) ◽  
pp. E609-E618
Author(s):  
Nebojsa N. Knezevic
Keyword(s):  
Polyethylene Glycol ◽  
Dorsal Root Ganglion ◽  
Sensory Neurons ◽  
Dorsal Root ◽  
Cytotoxic Effect ◽  
Caspase 3 ◽  
Tunel Assay ◽  
Methylprednisolone Acetate ◽  
Drg Neurons ◽  
Isolated Rat

Background: Epidural and intrathecal injections of methylprednisolone acetate (MPA) have become the most commonly performed interventional procedures in the United States and worldwide in the last 2 decades. However neuraxial MPA injection has been dogged by controversy regarding the presence of different additives used in commercially prepared glucocorticoids. We previously showed that MPA could be rendered 85% free of polyethylene glycol (PEG) by a simple physical separation of elements in the suspension. Objective: The objective of the present study was to explore a possible cytotoxic effect of commercially available MPA (with intact or reduced preservatives) on rat sensory neurons. Methods: We exposed primary dissociated rat dorsal root ganglia (DRG) sensory neurons to commercially available MPA for 24 hours with either the standard (commercial) concentration of preservatives or to different fractions following separation (MPA suspension whose preservative concentration had been reduced, or fractions containing higher concentrations of preservatives). Cells were stained with the TUNEL assay kit to detect apoptotic cells and images were taken on the Bio-Rad Laser Sharp-2000 system. We also detected expression of caspase-3, as an indicator of apoptosis in cell lysates. Results: We exposed sensory neurons from rat DRG to different concentrations of MPA from the original commercially prepared vial. TUNEL assay showed dose-related responses and increased percentages of apoptotic cells with increasing concentrations of MPA. Increased concentrations of MPA caused 1.5 – 2 times higher caspase-3 expression in DRG sensory neurons than in control cells (ANOVA, P = 0.001). Our results showed that MPA with reduced preservatives caused significantly less apoptosis observed with TUNEL assay labeling (P < 0.001) and caspase-3 immunoblotting (P ≤ 0.001) than in neurons exposed to MPA from a commercially prepared vial or “clear phase” that contained higher concentrations of preservatives. Even though MPA with reduced preservatives caused 12.5% more apoptosis in DRG sensory neurons than in control cells, post hoc analysis showed no differences between these 2 groups. Limitations: Our data was collected from in vitro isolated rat DRG neurons. There is a possibility that in vivo neurons have different extents of vulnerability compared to isolated neurons. Conclusions: Results of the present study identified a cytotoxic effect of commercially available MPA with preservatives or with a “clear phase” containing higher concentrations of preservatives on primary isolated rat DRG sensory neurons. This was shown by TUNEL positive assay and by increased caspase-3 expression as one of the final executing steps in apoptotic pathways in DRG neurons. However, our results showed no statistically significant difference between the control cells (salinetreated) and cells treated with MPA with reduced concentrations of preservatives, pointing out that either PEG or myristylgamma-picolinium chloride (MGPC) or their combination have harmful effects on these cells. Reduction of concentrations of preservatives from commercially available MPA suspensions by using the simple method of inverting vials for 2 hours could be considered useful in clinical practice to enhance the safety of this depot steroid when injected neuraxially. Key words: Methylprednisolone acetate, preservatives, dorsal root ganglion sensory neurons, cytotoxic effect, polyethylene glycol, myristylgamma-picolinium chloride

scholarly journals Spatial transcriptomics reveals unique molecular fingerprints of human nociceptors

2021 ◽  
Author(s):  
Diana Tavares-Ferreira ◽  
Stephanie Shiers ◽  
Pradipta R. Ray ◽  
Andi Wangzhou ◽  
Vivekanand Jeevakumar ◽  
...  
Keyword(s):  
Sex Differences ◽  
Sensory Neurons ◽  
Dorsal Root ◽  
Expression Profiles ◽  
Organ Donors ◽  
Drg Neurons ◽  
Pain Mechanisms ◽  
Molecular Fingerprints ◽  
Sensory Disorders

AbstractSingle-cell transcriptomics on mouse nociceptors has transformed our understanding of pain mechanisms. Equivalent information from human nociceptors is lacking. We used spatial transcriptomics to molecularly characterize transcriptomes of single dorsal root ganglion (DRG) neurons from 8 organ donors. We identified 10 clusters of human sensory neurons, 6 of which are C nociceptors, 1 Aβ nociceptor, 1 Aδ, and 2 Aβ subtypes. These neuron subtypes have distinct expression profiles from rodents and non-human primates and we identify new markers for each of these subtypes that can be applied broadly in human studies. We also identify sex differences, including a marked increase in CALCA expression in female putative itch nociceptors. Our data open the door to new pain targets and unparalleled molecular characterization of clinical sensory disorders.One Sentence SummaryThree A-fiber mechanoreceptor and seven nociceptor subtypes are identified, revealing sex differences and unique aspects of human DRG neurons.

Interaction of opioids and membrane potential to modulate Ca2+ channels in rat dorsal root ganglion neurons

1995 ◽  
Vol 73 (5) ◽  
pp. 1793-1798 ◽  
Author(s):  
M. D. Womack ◽  
E. W. McCleskey
Keyword(s):  
Dorsal Root Ganglion ◽  
Sensory Neurons ◽  
Action Potentials ◽  
Dorsal Root ◽  
Dorsal Root Ganglion Neurons ◽  
High Threshold ◽  
Partial Recovery ◽  
Drg Neurons ◽  
Ganglion Neurons ◽  
Ca2 Channels

1. Using patch-clamp methods, we show that brief prepulses to very positive voltages increase (facilitate) the amplitude of current through Ca2+ channels during a subsequent test pulse in some, but not all, dorsal root ganglion (DRG) sensory neurons. The amplitude of this facilitated current generally increases when the Ca2+ channels are inhibited by activation of the mu-opioid receptor. 2. The facilitated current is blocked by omega-conotoxin GVIA, activates in the range of high-threshold Ca2+ channels, and inactivates at relatively negative holding voltages. Thus facilitated current passes through N-type Ca2+ channels, the same channels that are inhibited by opioids and control neurotransmitter release in sensory neurons. 3. Although maximal facilitation occurs only at unphysiologically high membrane potentials (above +100 mV), some facilitation is seen after prepulses to voltages reached during action potentials. After return to the holding potential, facilitation persists for hundreds of milliseconds, considerably longer than in other neurons. Brief trains of pulses designed to mimic action potentials caused small facilitation (19% of maximal) in a fraction (8 of 24) of opioid-inhibited neurons. 4. We conclude that 1) prepulses to extremely positive voltages can cause partial recovery of Ca2+ channels inhibited by opioids; and 2) small, but detectable, facilitation is also seen after physiological stimulation in some DRG neurons. Facilitation, largely considered a biophysical epiphenomenon because of the extreme voltages used to induce it, appears to be physiologically relevant during opioid inhibition of Ca2+ channels in DRG neurons.

scholarly journals TNF-α acutely enhances acid-sensing ion channel currents in rat dorsal root ganglion neurons via a p38 MAPK pathway

2021 ◽  
Author(s):  
Wang-Ping Hu ◽  
Shuang Wei ◽  
Ying Jin ◽  
Ting-Ting Liu ◽  
Chun-Yu Qiu
Keyword(s):  
Ion Channels ◽  
Dorsal Root Ganglion ◽  
P38 Mapk ◽  
Sensory Neurons ◽  
Functional Activity ◽  
Dorsal Root ◽  
Mapk Pathway ◽  
Drg Neurons ◽  
P38 Mapk Pathway ◽  
Tnf Α

Abstract Background Tumor necrosis factor-α (TNF-α) is a pro-inflammatory cytokine involved in pain processing and hypersensitivity. It regulates not only the expression of a variety of inflammatory mediators but also the functional activity of some ion channels. Acid-sensing ion channels (ASICs), as key sensors for extracellular protons, are expressed in nociceptive sensory neurons and contribute to pain signaling caused by tissue acidosis. It is still unclear whether TNF-α has an effect on functional activity of ASICs. Herein, we reported that a brief exposure of TNF-α acutely sensitized ASICs in rat dorsal root ganglion (DRG) neurons. Methods Electrophysiological experiments were performed on neurons from rat DRG. Nociceptive behaviour was induced by acetic acid in rats. Results A brief (5min) application of TNF-α rapidly enhanced ASIC-mediated currents in rat DRG neurons. TNF-α (0.1-10 ng/ml) dose-dependently increased the proton-evoked ASIC currents with an EC 50 value of 1.96 ± 0.15 ng/ml. TNF-α shifted the concentration-response curve of proton upwards with a maximal current response increase of 42.34 ± 7.89%. In current-clamp recording, an acute application of TNF-α also significantly increased acid-evoked firing in rat DRG neurons. The rapid enhancement of ASIC-mediated electrophysiological activity by TNF-α was prevented by p38 mitogen-activated protein kinase (MAPK) inhibitor SB202190, but not by non-selective cyclooxygenase inhibitor indomethacin, suggesting that p38 MAPK is necessary for this enhancement. Behaviorally, TNF-α exacerbated acid-induced nociceptive behaviors in rats via activation of local p38 MAPK pathway. Conclusions These results suggest that TNF-α rapidly enhanced ASIC-mediated functional activity via a p38 MAPK pathway, which revealed a novel peripheral mechanism underlying TNF-α involvement in rapid hyperalgesia by sensitizing ASICs in primary sensory neurons.

Long-term IL-1β exposure causes subpopulation-dependent alterations in rat dorsal root ganglion neuron excitability

2012 ◽  
Vol 107 (6) ◽  
pp. 1586-1597 ◽  
Author(s):  
Patrick L. Stemkowski ◽  
Peter A. Smith
Keyword(s):  
Neuropathic Pain ◽  
Dorsal Root Ganglion ◽  
Sensory Neurons ◽  
Dorsal Root ◽  
Large Diameter ◽  
Small Diameter ◽  
Defined Medium ◽  
Drg Neurons ◽  
Enriched Cultures

The effect of interleukin-1β (IL-1β) on the electrical properties of sensory neurons was assessed at levels and exposure times comparable to those found in animal models of neuropathic pain. Experiments involved whole cell current-clamp recordings from rat dorsal root ganglion (DRG) neurons in defined-medium, neuron-enriched cultures. Five- to six-day exposure to 100 pM IL-1β produced subpopulation-dependent effects on DRG neurons. These included an increase in the excitability of medium-diameter and small-diameter isolectin B4 (IB4)-positive neurons that was comparable to that found after peripheral nerve injury. By contrast, a reduction in excitability was observed in large-diameter neurons, while no effect was found in small-diameter IB4-negative neurons. Further characterization of changes in medium and small IB4-positive neurons revealed that some, but not all, effects of IL-1β were mediated through its receptor, IL-1RI. Although the acute actions of IL-1β on sensory neurons have been well studied and related to acute and/or inflammatory pain, the present study shows how sensory neurons respond to long-term cytokine exposure. Such effects are relevant to understanding processes that contribute to the onset of neuropathic pain.

DRASIC Contributes to pH-Gated Currents in Large Dorsal Root Ganglion Sensory Neurons by Forming Heteromultimeric Channels

2002 ◽  
Vol 87 (6) ◽  
pp. 2835-2843 ◽  
Author(s):  
Jinghui Xie ◽  
Margaret P. Price ◽  
Allan L. Berger ◽  
Michael J. Welsh
Keyword(s):  
Dorsal Root Ganglion ◽  
Ion Channel ◽  
Sensory Neurons ◽  
Molecular Basis ◽  
Dorsal Root ◽  
Large Diameter ◽  
Wild Type ◽  
Drg Neurons ◽  
Whole Cell ◽  
Enhanced Sensitivity

For many years it has been observed that extracellular acid activates transient cation currents in large-diameter mechanosensory dorsal root ganglion (DRG) neurons. However, the molecular basis of these currents has not been known. Large DRG neurons express the dorsal root acid sensing ion channel (DRASIC), suggesting that DRASIC might contribute to H+-gated DRG currents. To test this, we examined whole cell currents in large DRG neurons from mice in which the DRASIC gene had been disrupted. We found that DRASIC null neurons retained H+-gated currents, indicating that DRASIC alone was not required for the currents. However, without DRASIC, the properties of the currents changed substantially as compared with wild-type neurons. In DRASIC –/– neurons, the rate of current desensitization in the continued presence of an acid stimulus slowed dramatically. H+-gated currents in DRASIC null neurons showed a decreased sensitivity to pH and an enhanced sensitivity to amiloride. The loss of DRASIC also altered but did not abolish the current potentiation generated by FMRF-related peptides. These data indicate that the DRASIC subunit makes an important contribution to H+-gated currents in large DRG sensory neurons. The results also suggest that related acid-activated DEG/ENaC channel subunits contribute with DRASIC to form heteromultimeric acid-activated channels.

GDNF and NGF Reverse Changes in Repriming of TTX-Sensitive Na+ Currents Following Axotomy of Dorsal Root Ganglion Neurons

2002 ◽  
Vol 88 (2) ◽  
pp. 650-658 ◽  
Author(s):  
Andreas Leffler ◽  
Theodore R. Cummins ◽  
Sulayman D. Dib-Hajj ◽  
William N. Hormuzdiar ◽  
Joel A. Black ◽  
...  
Keyword(s):  
Dorsal Root Ganglion ◽  
Sodium Channel ◽  
Sensory Neurons ◽  
Dorsal Root ◽  
Neuronal Excitability ◽  
Dorsal Root Ganglion Neurons ◽  
Current Profile ◽  
Drg Neurons ◽  
Α Subunit ◽  
Ganglion Neurons

Uninjured C-type rat dorsal root ganglion (DRG) neurons predominantly express slowly inactivating TTX-resistant (TTX-R) and slowly repriming TTX-sensitive (TTX-S) Na+ currents. After peripheral axotomy, TTX-R current density is reduced and rapidly repriming TTX-S currents emerge and predominate. The change in TTX-S repriming kinetics is paralleled by an increase in the level of transcripts and protein for the Nav1.3 sodium channel α-subunit, which is known to exhibit rapid repriming. Changes in Na+current profile and kinetics in DRG neurons may substantially alter neuronal excitability and could contribute to some states of chronic pain associated with injury of sensory neurons. In the present study, we asked whether glial-derived neurotrophic factor (GDNF) and nerve growth factor (NGF), which have been shown to prevent some axotomy-induced changes such as the loss of TTX-R Na+ current expression in DRG neurons, can ameliorate the axotomy-induced change in TTX-S Na+ current repriming kinetics. We show that intrathecally administered GDNF and NGF, delivered individually, can partially reverse the effect of axotomy on the repriming kinetics of TTX-S Na+ currents. When GDNF and NGF were co-administered, the repriming kinetics were fully rescued. We observed parallel effects of GDNF and NGF on the Nav1.3 sodium channel transcript levels in axotomized DRG. Both GDNF and NGF were able to partially reverse the axotomy-induced increase in Nav1.3 mRNA, with GDNF plus NGF producing the largest effect. Our data indicate that both GDNF and NGF can partially reverse an important effect of axotomy on the electrogenic properties of sensory neurons and that their effect is additive.

Expression of Galanin in Rat Primary Sensory Afferents After Moxibustion to the Skin

1992 ◽  
Vol 20 (02) ◽  
pp. 103-114 ◽  
Author(s):  
Hitoshi Kashiba ◽  
Ayahiko Nishigori ◽  
Yoshihiro Ueda
Keyword(s):  
Substance P ◽  
Dorsal Root Ganglion ◽  
Sensory Neurons ◽  
Untreated Control ◽  
Dorsal Root ◽  
Primary Sensory Neurons ◽  
Drg Neurons ◽  
Sensory Afferents ◽  
Retrograde Tracer ◽  
Control Skin

We examined the effects of moxibustion on primary sensory neurons in the skin of rats using immunocytochemistry combined with a fluorescent retrograde tracer dye, fluoro gold (FG). Galanin-like immunoreactive (IR) fibers were often observed in the dermis of treated skin at 18 hours after moxibustion, while such fibers were rarely detected in untreated (control) skin. Moreover, most of galanin-IR fibers also displayed substance P(SP)-like immunoreactivity. About 20-30% of the dorsal root ganglion (DRG) neurons labeled when FG was injected intradermally into the moxibustion-treated skin showed galanin-like immunoreactivity, while the proportion of FG-labeled neurons with such immunoreactivity was < 10% in control DRGs. These results show that moxibustion induced galanin expression by primary sensory neurons containing SP. The possible functions of this peptide are discussed in relation to the effects of moxibustion.

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