Spinal Nerve Ligation
Objective: To assess neuropathic pain-like behavioral changes induced by spinal nerve ligation and evaluate the role of caspase-dependent apoptosis in dorsal horn neurons
This is a Spinal Nerve Ligation protocol using Not explicitly stated in provided text as the model organism. The procedure involves 6 procedural steps, 1 equipment items, 1 materials. Extracted from a 2005 paper published in Journal of Neuroscience.
Model and subjects
Not explicitly stated in provided text • Not explicitly stated in provided text • unknown • Not explicitly stated in provided text • Not explicitly stated in provided text
Study window
Estimated timing pending
Core workflow
Perform spinal nerve ligation • Allow time for apoptosis induction • Administer caspase inhibitor treatment
Primary readouts
- Cumulative loss of dorsal horn neurons (>20% at four weeks)
- Loss of GABAergic inhibitory interneurons
- Reduction of inhibitory postsynaptic currents in lamina II neurons
- Hypersensitivity to tactile stimuli
Key equipment and reagents
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Protocol Steps
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Perform spinal nerve ligation
Conduct partial peripheral nerve lesion procedure using spinal nerve ligation technique
Note: This is one of three distinct partial peripheral nerve lesion models used in the study
View evidence from paper
“three distinct partial peripheral nerve lesions: spared nerve injury, chronic constriction, and spinal nerve ligation”
Allow time for apoptosis induction
Wait for transsynaptic apoptosis to be induced in superficial dorsal horn laminas I-III
Note: Apoptosis is slow in onset and persists for several weeks
View evidence from paper
“a caspase-dependent degeneration of dorsal horn neurons that is slow in onset and persists for several weeks”
Administer caspase inhibitor treatment
Treat animals with zVAD caspase inhibitor to block apoptosis
Note: zVAD has no intrinsic analgesic properties
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 measures response 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”
Perform stereological analysis of dorsal horn neurons
Quantitatively analyze cumulative loss of dorsal horn neurons at four weeks post-injury
Note: Analysis determines >20% cumulative neuron loss after spared nerve injury
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
Assess inhibitory postsynaptic currents in lamina II neurons to evaluate GABAergic transmission
Note: Marked decrease in inhibitory 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”