Experiments/Spinal Nerve Ligation

Source Paper

Blocking Caspase Activity Prevents Transsynaptic Neuronal Apoptosis and the Loss of Inhibition in Lamina II of the Dorsal Horn after Peripheral Nerve Injury

Joachim Scholz, Daniel C. Broom, Dong-Ho Youn, Charles D. Mills, Tatsuro Kohno et al.

Journal of Neuroscience • 2005

DOI PMC

Spinal Nerve Ligation

behavioralNot explicitly stated in provided textNot explicitly stated in provided text

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

Materials & Equipment Checklist

2 items

Gather these items before starting the experiment. Check off items as you prepare.

Equipment1

Stereological analysis equipment

Not explicitly stated • Not explicitly stated • Not explicitly stated • Not explicitly stated

Amazon

Materials1

Caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone

Not explicitly stated • Not explicitly stated • Not explicitly stated • Not explicitly stated

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Protocol Steps

Perform spinal nerve ligation

Conduct partial peripheral nerve lesion procedure using spinal nerve ligation technique

Not explicitly statedNot explicitly stated

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

Several weeks, with analysis at four weeks post-injuryNot explicitly stated

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

Not explicitly statedNot explicitly stated

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

Not explicitly statedNot explicitly stated

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

Four weeks post-injuryNot explicitly stated

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

Not explicitly statedNot explicitly stated

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”