Behavioral Pain Testing
Objective: Assessment of pain-like behavioral changes including hypersensitivity to tactile or cold stimuli following peripheral nerve injury, and evaluation of caspase inhibitor treatment effects on neuropathic pain
Gather these items before starting the experiment. Check off items as you prepare.
Equipment1
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Materials1
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Protocol Steps
Peripheral nerve injury induction
Three distinct partial peripheral nerve lesions were performed: spared nerve injury, chronic constriction, and spinal nerve ligation
Note: Multiple nerve injury models were used to induce transsynaptic apoptosis in the superficial dorsal horn
View evidence from paper
“We show that transsynaptic apoptosis is induced in the superficial dorsal horn (laminas I-III) of the spinal cord by three distinct partial peripheral nerve lesions: spared nerve injury, chronic constriction, and spinal nerve ligation”
Assessment of pain-like behavioral changes
Evaluation of hypersensitivity to tactile or cold stimuli following peripheral nerve injury
Note: Behavioral testing measured pain-like responses to tactile and cold stimuli
View evidence from paper
“Partial peripheral nerve injury results in pain-like behavioral changes characterized by hypersensitivity to tactile or cold stimuli”
Treatment with caspase inhibitor
Administration of zVAD caspase inhibitor to block apoptosis
Note: zVAD has no intrinsic analgesic properties but attenuates neuropathic pain-like syndrome
View evidence from paper
“Treatment with zVAD, which has no intrinsic analgesic properties, attenuates this neuropathic pain-like syndrome”
Neuronal loss quantification
Stereological analysis performed four weeks after spared nerve injury to determine cumulative loss of dorsal horn neurons
Note: Loss of dorsal horn neurons exceeded 20% at this timepoint
View evidence from paper
“Four weeks after spared nerve injury, the cumulative loss of dorsal horn neurons, determined by stereological analysis, is >20%”
Electrophysiological assessment
Measurement of inhibitory postsynaptic currents in lamina II neurons to assess GABAergic transmission
Note: Marked decrease in inhibitory postsynaptic currents coincided with apoptosis induction
View evidence from paper
“a marked decrease in inhibitory postsynaptic currents of lamina II neurons coincides with the induction of apoptosis”