Proteomic identification of brainstem cytosolic proteins in a neuropathic pain model.
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.
Alzate
Oscar
O
Neuroproteomic Center, Department of Neurobiology, Duke University Medical Center, Durham, NC, USA.
Hussain
Syed-Rehan A
SR
Goettl
Virginia M
VM
Tewari
Arun K
AK
Madiai
Francesca
F
Stephens
Robert L
RL
Hackshaw
Kevin V
KV
D000818:Animals; D001933:Brain Stem / Q000473:pathology*; D003600:Cytosol / Q000378:metabolism; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D008026:Ligation / Q000379:methods; D008297:Male; D009437:Neuralgia / Q000378:metabolism*; D010147:Pain Measurement / Q000379:methods; D010449:Peptide Mapping / Q000379:methods; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013127:Spinal Nerves / Q000378:metabolism*
D011506:Proteins