Gel-based proteomics of unilateral irradiated striatum after gamma knife surgery.
Gamma knife surgery (GKS) is used for the treatment of various brain disorders. The biological effects of focal gamma ray irradiation on targeted or surrounding areas in the brain are not well-known. In the present study, we evaluated protein expression changes in the unilateral irradiated (60 Gy) striatum in rat. Striata of irradiated and control brains were dissected 16 h post-irradiation for analysis by large-format two-dimensional gel electrophoresis (2-DGE). In parallel, we also examined the un-targeted contralateral striatum over the control for potential changes in proteins patterns that may have occurred due to the effects of irradiation to the unilateral striatum. A total of 17 reproducible and differentially expressed silver nitrate-stained protein spots in the irradiated striatum was detected on 2-D gel. Their subsequent analysis by tandem mass spectrometry (nESI-LC-MS/MS) resulted in the identification of 13 nonredundant proteins. Interestingly, out of these 13 changed proteins, 2 proteins were also detected in the contralateral striatum. Some of the significantly changed proteins identified were creatine kinase, protein disulfide isomerase A3 precursor (PDA3), and peroxiredoxin 2 (Prx2). Western analysis with anti-PDA3 and anti-Prx2 antibodies revealed 4 and 2 cross-reacting protein spots on 2-D gel blots. Interestingly, after GKS, in the irradiated and un-irradiated striata, these spots showed a shift toward the acidic side, suggesting post-translational modifications. Taken together, these results indicate that unilateral irradiation during GKS triggers molecular changes in the bilateral striata.
Hirano
Misato
M
Human Stress Signal Research Center (HSS), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba West, 16-1 Onogawa, Tsukuba 305-8569, Japan.
Rakwal
Randeep
R
Kouyama
Nobuo
N
Katayama
Yoko
Y
Hayashi
Motohiro
M
Shibato
Junko
J
Ogawa
Yoko
Y
Yoshida
Yasukazu
Y
Iwahashi
Hitoshi
H
Masuo
Yoshinori
Y
D000595:Amino Acid Sequence; D000818:Animals; D003342:Corpus Striatum / Q000737:chemistry* / Q000528:radiation effects* / Q000601:surgery; D003593:Cytoplasm / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism; D015870:Gene Expression / Q000187:drug effects; D008297:Male; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis; D040901:Proteomics; D016634:Radiosurgery; D051381:Rats; D011920:Rats, Inbred WF / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013482:Superoxide Dismutase / Q000032:analysis; D000072105:Superoxide Dismutase-1; D053719:Tandem Mass Spectrometry
D009419:Nerve Tissue Proteins; C000606290:SOD1 protein, human; C000606292:Sod1 protein, rat; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1