Musculoskeletal pain resulting from tissue ischemia with reperfusion is a major health issue that affects millions of people in the United States. Peripheral ischemia/ reperfusion occurs in blood disorders like sickle cell disease, and in cardiovascular disorders such as peripheral vascular disease. Ischemia/ reperfusion are also thought to be the underlying cause of complex regional pain syndrome. While much is known about the functional properties and plasticity of cutaneous nociceptors following peripheral injuries and how these fibers contribute to pain, relatively little is known about the functional properties of group III and IV muscle afferents and their role in muscle pain development. The major goal of this proposal is to determine the molecular mechanisms of muscle afferent sensitization that may underlie muscle pain during ischemia and after tissue reperfusion. We hypothesize that these distinct phases cause differential changes in heat, mechanical and chemo-sensitivity in muscle afferents, which are mediated by upregulation of purinergic receptors during ischemia and acid sensing ion channels after reperfusion leading to muscle pain. In order to increase our knowledge of muscle afferents, we developed a novel ex vivo forepaw muscle, median & ulnar nerves, dorsal root ganglion (DRG), spinal cord recording preparation that enables us to comprehensively phenotype these afferents in mouse. We are also able to analyze the central anatomy, and the neurochemical or molecular phenotypes of these afferents using combinations of ex vivo recording with immunocytochemical and single cell RT-PCR analyses.
In Specific Aim 1, we will determine if upregulation of purinergic receptors, P2Y1 and P2X5, regulate the observed changes in heat and chemosensitivity in muscle afferents, respectively during ischemia using in vivo siRNA-mediated knockdown of these genes in single peripheral nerves in conjunction with ex vivo recording preparations. Next, in Specific Aim 2, we will utilize a similar approach to SA1 except we will determine if upregulation of ASIC1 and ASIC3 regulate the novel changes in mechanical and metabolite responses in muscle afferents, respectively after transient ischemia with tissue reperfusion. Finally, in Specific Aim 3, we will determine if upregulation of P2Y1 and P2X5 regulate muscle pain during ischemia while upregulation of ASIC1 and ASIC3 regulate muscle pain after reperfusion by analyzing the effects of receptor knockdown on recognized muscle pain behavior tests between these two phases. This study will enable us to characterize the changes in both non-nociceptive and nociceptive muscle afferents after ischemia/ reperfusion and identify unique mechanisms associated with muscle afferent sensitization that underlie muscle pain development. This may lead to the formulation of more appropriate treatments for musculoskeletal pain associated with ischemia/ reperfusion that target the proper pain receptor(s) or primary afferent subpopulation(s).

Public Health Relevance

The results of the proposed experiments will significantly enhance our understanding of the molecular mechanisms underlying sensory neuron responses and hopefully provide novel information for use in the development of new avenues of research and therapies for musculoskeletal pain and/ or altered cardiovascular reflexes associated with ischemia/ reperfusion. The information obtained from these novel studies will allow us to better understand the functional implications of sensitization in specific subpopulations of muscle sensory neurons and the role they play in the development of pain states.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
4R01AR064551-04
Application #
9132597
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Witter, James
Project Start
2013-09-17
Project End
2018-08-31
Budget Start
2016-09-01
Budget End
2017-08-31
Support Year
4
Fiscal Year
2016
Total Cost
$325,125
Indirect Cost
$112,625
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
He, Xuelian; Zhang, Liguo; Queme, Luis F et al. (2018) A histone deacetylase 3-dependent pathway delimits peripheral myelin growth and functional regeneration. Nat Med 24:338-351
Ross, Jessica L; Queme, Luis F; Lamb, Jordan E et al. (2018) Interleukin 1? inhibition contributes to the antinociceptive effects of voluntary exercise on ischemia/reperfusion-induced hypersensitivity. Pain 159:380-392
Coover, Robert A; Healy, Tabitha E; Guo, Li et al. (2018) Tonic ATP-mediated growth suppression in peripheral nerve glia requires arrestin-PP2 and is evaded in NF1. Acta Neuropathol Commun 6:127
Ross, Jessica L; Queme, Luis F; Lamb, Jordan E et al. (2018) Sex differences in primary muscle afferent sensitization following ischemia and reperfusion injury. Biol Sex Differ 9:2
Lu, Peilin; Hudgins, Renita C; Liu, Xiaohua et al. (2017) Upregulation of P2Y1 in neonatal nociceptors regulates heat and mechanical sensitization during cutaneous inflammation. Mol Pain 13:1744806917730255
Li, Zhuo; Peng, Yanyan; Hufnagel, Robert B et al. (2017) Loss of SLC25A46 causes neurodegeneration by affecting mitochondrial dynamics and energy production in mice. Hum Mol Genet 26:3776-3791
Queme, Luis F; Ross, Jessica L; Jankowski, Michael P (2017) Peripheral Mechanisms of Ischemic Myalgia. Front Cell Neurosci 11:419
Ross, Jessica L; Queme, Luis F; Cohen, Elysia R et al. (2016) Muscle IL1? Drives Ischemic Myalgia via ASIC3-Mediated Sensory Neuron Sensitization. J Neurosci 36:6857-71
Wu, Lai Man Natalie; Wang, Jincheng; Conidi, Andrea et al. (2016) Zeb2 recruits HDAC-NuRD to inhibit Notch and controls Schwann cell differentiation and remyelination. Nat Neurosci 19:1060-72
Queme, Luis F; Ross, Jessica L; Lu, Peilin et al. (2016) Dual Modulation of Nociception and Cardiovascular Reflexes during Peripheral Ischemia through P2Y1 Receptor-Dependent Sensitization of Muscle Afferents. J Neurosci 36:19-30

Showing the most recent 10 out of 12 publications