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Protocol Steps
Start here. The step list is optimized for running the experiment, with direct vendor links available inline when you need to source a cited item.
Spinal Nerve Ligation Surgery
Perform unilateral lumbar spinal nerve ligation on rats to create an in vivo model of neuropathy
Note: This is the primary surgical intervention to induce neuropathic pain and allodynia in the animal model
View evidence from paper
“we used an in vivo rat model of neuropathy, unilateral lumbar spinal nerve ligation (SNL), to characterize the distribution of α2δ-1”
Characterize α2δ-1 Distribution
Examine the distribution of α2δ-1 in DRG neurons at light- and electron-microscopic levels on the ligated side
Note: Analysis includes endoplasmic reticulum of DRG somata, intracellular vesicular structures within axons, and plasma membrane of presynaptic terminals in dorsal horn superficial layers
View evidence from paper
“to characterize the distribution of α2δ-1 in DRG neurons, both at the light- and electron-microscopic level”
Chronic Pregabalin Treatment
Administer chronic pregabalin (PGB) treatment to SNL animals at a dose that alleviates allodynia
Note: Treatment dose selected based on its ability to alleviate allodynia symptoms
View evidence from paper
“Chronic PGB treatment of SNL animals, at a dose that alleviated allodynia”
Assess α2δ-1 Changes in Spinal Cord
Measure elevation of α2δ-1 in the spinal cord and ascending axon tracts following chronic pregabalin treatment
Note: Pregabalin markedly reduced the elevation of α2δ-1 in these regions
View evidence from paper
“markedly reduced the elevation of α2δ-1 in the spinal cord and ascending axon tracts”
Assess α2δ-1 Changes in DRGs
Measure α2δ-1 mRNA and protein upregulation in dorsal root ganglia following chronic pregabalin treatment
Note: Pregabalin had no effect on the upregulation of α2δ-1 mRNA and protein in DRGs
View evidence from paper
“it had no effect on the upregulation of α2δ-1 mRNA and protein in DRGs”
In Vitro Analysis of Pregabalin Effects
Conduct in vitro experiments to determine pregabalin's effects on plasma membrane expression of α2δ-1 and endocytosis
Note: Pregabalin reduced plasma membrane expression of α2δ-1 without affecting endocytosis
View evidence from paper
“In vitro, PGB reduced plasma membrane expression of α2δ-1 without affecting endocytosis”