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Quantitative proteomic analysis of cellular nuclear protein TMT markers from rat spinal cord tissue samples in neuropathic pain model

ABSTRACT: To further reveal the mechanism of neuropathic pain, we simulated neuropathic pain by constructing a chronic constriction injury of the sciatic nerve (CCI) model, followed by quantitative proteomic analysis of rat spinal cord cell nuclei at 1 day, 7 days and sham-operated groups after CCI. A total of 3 samples from each group were used. A total of 5039 proteins were identified in this study, of which 4469 proteins contained quantitative information. Differential proteins were defined by a threshold of change >1.3-fold and t-test p-value <0.05.v

INSTRUMENT(S): Q Exactive

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Spinal Cord

SUBMITTER: Zhifeng Huang

LAB HEAD: Changsheng Huang

PROVIDER: PXD034960 | Pride | 2023-01-17

REPOSITORIES: Pride

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Project description:Not all patients with nerve injury develop neuropathic pain. The extent of nerve damage and age at the time of injury are two of the few risk factors identified to date. In addition, preclinical studies show that neuropathic pain variance is heritable. To define such factors further, we performed a large-scale gene profiling experiment which plotted global expression changes in the rat dorsal root ganglion in three peripheral neuropathic pain models. This resulted in the discovery that the potassium channel alpha subunit KCNS1, involved in neuronal excitability, is constitutively expressed in sensory neurons and markedly downregulated following nerve injury. KCNS1 was then characterized by an unbiased network analysis as a putative pain gene, a result confirmed by single nucleotide polymorphism association studies in humans. A common amino acid changing allele, the 'valine risk allele', was significantly associated with higher pain scores in five of six independent patient cohorts assayed (total of 1359 subjects). Risk allele prevalence is high, with 18-22% of the population homozygous, and an additional 50% heterozygous. At lower levels of nerve damage (lumbar back pain with disc herniation) association with greater pain outcome in homozygote patients is P = 0.003, increasing to P = 0.0001 for higher levels of nerve injury (limb amputation). The combined P-value for pain association in all six cohorts tested is 1.14 E-08. The risk profile of this marker is additive: two copies confer the most, one intermediate and none the least risk. Relative degrees of enhanced risk vary between cohorts, but for patients with lumbar back pain, they range between 2- and 3-fold. Although work still remains to define the potential role of this protein in the pathogenic process, here we present the KCNS1 allele rs734784 as one of the first prognostic indicators of chronic pain risk. Screening for this allele could help define those individuals prone to a transition to persistent pain, and thus requiring therapeutic strategies or lifestyle changes that minimize nerve injury. Microarrays were run on mRNA extracted from adult rat L4 and L5 DRGs cells after 3,7,21,40 hours after three different sciatic nerve lesions [Spared Nerve Injury (SNI); Chronic Constriction Injury (CCI); Spinal Nerve Ligation (Ch) with Sham controls (SH)].

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Quantitative proteomic analysis of cellular nuclear protein TMT markers from rat spinal cord tissue samples in neuropathic pain model

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