Spared Nerve Injury
Objective: To assess transsynaptic apoptosis in the dorsal horn following spared nerve injury and evaluate resulting neuropathic pain-like behavioral changes, including hypersensitivity to tactile or cold stimuli
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Materials1
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Protocol Steps
Spared Nerve Injury Surgery
Perform spared nerve injury as a partial peripheral nerve lesion to induce transsynaptic apoptosis in the dorsal horn
Note: This is one of three distinct partial peripheral nerve lesions tested; others include chronic constriction and spinal nerve ligation
View evidence from paper
“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”
Assess Neuronal Loss via Stereological Analysis
Determine cumulative loss of dorsal horn neurons using stereological analysis at four weeks post-injury
Note: Cumulative loss of dorsal horn neurons exceeds 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%”
Measure Inhibitory Postsynaptic Currents
Record inhibitory postsynaptic currents from lamina II neurons to assess GABAergic transmission changes
Note: A marked decrease in inhibitory postsynaptic currents coincides with apoptosis induction
View evidence from paper
“a marked decrease in inhibitory postsynaptic currents of lamina II neurons coincides with the induction of apoptosis”
Administer Caspase Inhibitor Treatment
Treat animals with zVAD caspase inhibitor to block apoptosis
Note: zVAD has no intrinsic analgesic properties but prevents loss of GABAergic interneurons and reduction of inhibitory currents
View evidence from paper
“Treatment with zVAD, which has no intrinsic analgesic properties, attenuates this neuropathic pain-like syndrome”
Assess Neuropathic Pain-like Behavioral Changes
Evaluate pain-like behavioral changes characterized by hypersensitivity to tactile or cold stimuli
Note: Behavioral testing assesses response to both 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”