Source Paper
Activated Microglia Contribute to the Maintenance of Chronic Pain after Spinal Cord Injury
Bryan C. Hains, Stephen G. Waxman
Journal of Neuroscience • 2006
Source Paper
Bryan C. Hains, Stephen G. Waxman
Journal of Neuroscience • 2006
Traumatic spinal cord injury (SCI) results not only in motor impairment but also in chronic central pain, which can be refractory to conventional treatment approaches. It has been shown recently that in models of peripheral nerve injury, spinal cord microglia can become activated and contribute to development of pain. Considering their role in pain after peripheral injury, and because microglia are known to become activated after SCI, we tested the hypothesis that activated microglia contribute to chronic pain after SCI. In this study, adult male Sprague Dawley rats underwent T9 spinal cord contusion injury. Four weeks after injury, when lumbar dorsal horn multireceptive neurons became hyperresponsive and when behavioral nociceptive thresholds were decreased to both mechanical and thermal stimuli, intrathecal infusions of the microglial inhibitor minocycline were initiated. Electrophysiological experiments showed that minocycline rapidly attenuated hyperresponsiveness of lumbar dorsal horn neurons. Behavioral data showed that minocycline restored nociceptive thresholds, at which time spinal microglial cells assumed a quiescent morphological phenotype. Levels of phosphorylated-p38 were decreased in SCI animals receiving minocycline. Cessation of delivery of minocycline resulted in an immediate return of pain-related phenomena. These results suggest an important role for activated microglia in the maintenance of chronic central below-level pain after SCI and support the newly emerging role of non-neuronal immune cells as a contributing factor in post-SCI pain.
Objective: To test the hypothesis that activated microglia contribute to chronic pain after spinal cord injury by inducing T9 spinal cord contusion injury and evaluating the effects of microglial inhibition on pain-related behaviors and neuronal hyperresponsiveness
This is a Spinal Cord Contusion Injury protocol using rat as the model organism. The procedure involves 9 procedural steps, 4 equipment items, 1 materials. Extracted from a 2006 paper published in Journal of Neuroscience.
Model and subjects
rat • Sprague Dawley • male • adult
Study window
~4 week study window
Core workflow
Spinal cord contusion injury induction • Post-injury observation period • Baseline behavioral assessment
Primary readouts
Key equipment and reagents
Verified items
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Adult male Sprague Dawley rats underwent T9 spinal cord contusion injury to model traumatic spinal cord injury
Note: Injury performed at thoracic level 9 (T9)
“adult male Sprague Dawley rats underwent T9 spinal cord contusion injury”
Animals were observed for four weeks after injury to allow development of chronic pain phenotype and microglial activation
Note: At this timepoint, lumbar dorsal horn neurons became hyperresponsive and behavioral nociceptive thresholds were decreased
“Four weeks after injury, when lumbar dorsal horn multireceptive neurons became hyperresponsive and when behavioral nociceptive thresholds were decreased to both mechanical and thermal stimuli”
Measurement of nociceptive thresholds to mechanical and thermal stimuli before minocycline treatment initiation
Note: Establishes baseline pain-related behavior at 4 weeks post-injury
“behavioral nociceptive thresholds were decreased to both mechanical and thermal stimuli”
Intrathecal infusions of minocycline microglial inhibitor were initiated at four weeks post-injury
Note: Delivery method and dosage not specified in provided text
“intrathecal infusions of the microglial inhibitor minocycline were initiated”
Electrophysiological experiments conducted to measure lumbar dorsal horn neuronal responses during minocycline administration
Note: Minocycline rapidly attenuated hyperresponsiveness of lumbar dorsal horn neurons
“Electrophysiological experiments showed that minocycline rapidly attenuated hyperresponsiveness of lumbar dorsal horn neurons”
Measurement of nociceptive thresholds to mechanical and thermal stimuli during minocycline infusion
Note: Minocycline restored nociceptive thresholds to normal levels
“Behavioral data showed that minocycline restored nociceptive thresholds”
Examination of spinal microglial cell morphology during minocycline treatment
Note: Microglial cells assumed a quiescent morphological phenotype during minocycline treatment
“spinal microglial cells assumed a quiescent morphological phenotype”
Measurement of phosphorylated-p38 protein levels in spinal cord tissue during minocycline treatment
Note: Levels of phosphorylated-p38 were decreased in SCI animals receiving minocycline
“Levels of phosphorylated-p38 were decreased in SCI animals receiving minocycline”
Discontinuation of minocycline intrathecal infusion to assess reversibility of treatment effects
Note: Cessation resulted in immediate return of pain-related phenomena
“Cessation of delivery of minocycline resulted in an immediate return of pain-related phenomena”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
To test the hypothesis that activated microglia contribute to chronic pain after spinal cord injury by inducing T9 spinal cord contusion injury and evaluating the effects of microglial inhibition on pain-related behaviors and neuronal hyperresponsiveness
Objective
To test the hypothesis that activated microglia contribute to chronic pain after spinal cord injury by inducing T9 spinal cord contusion injury and evaluating the effects of microglial inhibition on pain-related behaviors and neuronal hyperresponsiveness
Subjects
From paperrat • Sprague Dawley • male • adult
Spinal cord contusion injury induction
Post-injury observation period (Four weeks)
Baseline behavioral assessment
Intrathecal minocycline infusion initiation
Nociceptive thresholds to mechanical stimuli
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Nociceptive thresholds to thermal stimuli
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Lumbar dorsal horn neuronal hyperresponsiveness (electrophysiological recording)
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Spinal microglial cell morphology (quiescent vs activated phenotype)
From paperNot specified in provided text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Nociceptive thresholds to mechanical stimuli
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Nociceptive thresholds to thermal stimuli
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Lumbar dorsal horn neuronal hyperresponsiveness (electrophysiological recording)
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Spinal microglial cell morphology (quiescent vs activated phenotype)
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
Not specified in provided text
Scoring or quantification
Quantify the primary readouts for this experiment: Nociceptive thresholds to mechanical stimuli; Nociceptive thresholds to thermal stimuli; Lumbar dorsal horn neuronal hyperresponsiveness (electrophysiological recording); Spinal microglial cell morphology (quiescent vs activated phenotype).
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Nociceptive thresholds to mechanical stimuli, Nociceptive thresholds to thermal stimuli, Lumbar dorsal horn neuronal hyperresponsiveness (electrophysiological recording), Spinal microglial cell morphology (quiescent vs activated phenotype).
Source links and direct wording from the methods section for validation and deeper review.
Citation
Bryan C. Hains et al. (2006). Activated Microglia Contribute to the Maintenance of Chronic Pain after Spinal Cord Injury. Journal of Neuroscience
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