Proteomic identification of brainstem cytosolic proteins in a neuropathic pain model

• Published in: Brain research. Molecular brain research. Vol. 128; no. 2; pp. 193 - 200
• Main Authors: Alzate, Oscar, Hussain, Syed-Rehan A., Goettl, Virginia M., Tewari, Arun K., Madiai, Francesca, Stephens, Robert L., Hackshaw, Kevin V.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 28.09.2004; Elsevier
• Subjects: 2-DE; Allodynia; Animals; Biological and medical sciences; Brain Stem - pathology; Cranial nerves. Spinal roots. Peripheral nerves. Autonomic nervous system. Gustation. Olfaction; Cytosol - metabolism; Disease Models, Animal; Electrophoresis, Gel, Two-Dimensional - methods; Fundamental and applied biological sciences. Psychology; Gas Chromatography-Mass Spectrometry - methods; Ligation - methods; MALDI-TOF; Male; Medical sciences; Nervous system (semeiology, syndromes); Neuralgia - metabolism; Neurology; Nociception; Pain Measurement - methods; Peptide Mapping - methods; Proteins - metabolism; Proteomics - methods; Rats; Rats, Sprague-Dawley; Somesthesis and somesthetic pathways (proprioception, exteroception, nociception); interoception; electrolocation. Sensory receptors; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Spinal nerve ligation; Spinal Nerves - metabolism; Vertebrates: nervous system and sense organs; Spinal nerve ligation; Nociception; Allodynia; Sensory systems; 2-DE; MALDI-TOF; Nervous system diseases; Brain stem; Central nervous system; Peripheral neuropathy; Spinal nerve; Encephalon; Proteomics; Models; Peripheral nerve disease
• ISSN: 0169-328X; 1872-6941
• DOI: 10.1016/j.molbrainres.2004.06.037

Neuropathic pain involves co-regulation of many genes and their translational products in both peripheral and central nervous system. We used proteomics approaches to investigate expressional changes in cytosolic protein levels in rat brainstem tissues following ligation of lumbar 5 and 6 (L5, L6) spinal nerves, which generates a model of peripheral neuropathic pain (NP). Proteins from brainstem tissue homogenates of NP and SHAM animals were fractionated by two-dimensional (2-DE) gel electrophoresis to produce a high-resolution map of the brainstem soluble proteins. Proteins showing altered expression levels between NP and SHAM were selected. Isolated proteins were in-gel trypsin-digested and the resulting peptides were analyzed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Using the mass spectrometric data, we were able to identify 17 proteins of interest through searches of the Swiss-Prot and NCBi nonredundant protein sequence database. Several of the identified proteins, including fatty acid binding protein-brain (FABP-B), major histocompatibility complex (MHC) class 1, T-cell receptor (TCR) alpha chain, and interleukin-1 (IL-1), showed significantly higher levels in the NP rat brainstem. Proteomic analysis has identified several proteins with differential expression levels in NP as compared to SHAM. However, the function of the proteins identified is postulated; therefore, further experiments are required to determine the true role of each protein in NP.