Modeling cerebral ischemia in neuroproteomics.
Protein changes induced by traumatic or ischemic brain injury can serve as diagnostic markers as well as therapeutic targets for neuroprotection. The focus of this chapter is to provide a representative overview of preclinical brain injury and proteomics analysis protocols for evaluation and discovery of novel biomarkers. Detailed surgical procedures have been provided for inducing MCAo and implantation of chronic indwelling cannulas for drug delivery. Sample collection and tissue processing techniques for collection of blood, CSF, and brain are also described including standard biochemical methodology for the proteomic analysis of these tissues.The dynamics of proteomic analysis is a multistep process comprising sample preparation, separation, quantification, and identification of proteins. Our approach is to separate proteins first by two-dimensional gel electrophoresis according to charge and molecular mass. Proteins are then fragmented and analyzed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Identification of proteins can be achieved by comparing the mass-to-charge data to protein sequences in respective databases.
Dave
Jitendra R
JR
Department of Applied Neurobiology, Walter Reed Army Institute of Research, Silver Spring, MD, USA. jit.dave@na.amedd.army.mil
Williams
Anthony J
AJ
Yao
Changping
C
Lu
X-C May
XC
Tortella
Frank C
FC
D000818:Animals; D001921:Brain / Q000473:pathology / Q000502:physiology; D002545:Brain Ischemia / Q000503:physiopathology*; D006801:Humans; D020244:Infarction, Middle Cerebral Artery; D009419:Nerve Tissue Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D051381:Rats
D009419:Nerve Tissue Proteins; D020543:Proteome