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Transcriptomics and proteomics analyses of the PACAP38 influenced ischemic brain in permanent middle cerebral artery occlusion model mice
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نویسنده
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hori m. ,nakamachi t. ,rakwal r. ,shibato j. ,ogawa t. ,aiuchi t. ,tsuruyama t. ,tamaki k. ,shioda s.
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منبع
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journal of neuroinflammation - 2012 - دوره : 9 - شماره : 0
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چکیده
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Introduction: the neuropeptide pituitary adenylate cyclase-activating polypeptide (pacap) is considered to be a potential therapeutic agent for prevention of cerebral ischemia. ischemia is a most common cause of death after heart attack and cancer causing major negative social and economic consequences. this study was designed to investigate the effect of pacap38 injection intracerebroventrically in a mouse model of permanent middle cerebral artery occlusion (pmcao) along with corresponding sham control that used 0.9% saline injection.methods: ischemic and non-ischemic brain tissues were sampled at 6 and 24 hours post-treatment. following behavioral analyses to confirm whether the ischemia has occurred,we investigated the genome-wide changes in gene and protein expression using dna microarray chip (4x44k,agilent) and two-dimensional gel electrophoresis (2-dge) coupled with matrix assisted laser desorption/ionization-time of flight-mass spectrometry (maldi-tof-ms),respectively. western blotting and immunofluorescent staining were also used to further examine the identified protein factor.results: our results revealed numerous changes in the transcriptome of ischemic hemisphere (ipsilateral) treated with pacap38 compared to the saline-injected sham control hemisphere (contralateral). previously known (such as the interleukin family) and novel (gabra6,crtam) genes were identified under pacap influence. in parallel,2-dge analysis revealed a highly expressed protein spot in the ischemic hemisphere that was identified as dihydropyrimidinase-related protein 2 (dpyl2). the dpyl2,also known as crmp2,is a marker for the axonal growth and nerve development. interestingly,pacap treatment slightly increased its abundance (by 2-dge and immunostaining) at 6 h but not at 24 h in the ischemic hemisphere,suggesting pacap activates neuronal defense mechanism early on.conclusions: this study provides a detailed inventory of pacap influenced gene expressions and protein targets in mice ischemic brain,and suggests new targets for thereaupetic interventions. © 2012 hori et al.; licensee biomed central ltd.
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کلیدواژه
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CRMP2; Crtam; DNA microarray; Gabra6; Il6; Ischemia; Neuroprotection; PACAP
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آدرس
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department of forensic medicine and molecular pathology,school of medicine,kyoto university,kyoto,606-8315,japan,department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555, Japan, department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555,japan,department of center for biotechnology,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555, Japan, department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555,japan,graduate school of life and environmental sciences,university of tsukuba,tsukuba,305-8572, Japan, department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555, Japan, department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555, Japan, department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555, Japan, department of forensic medicine and molecular pathology,school of medicine,kyoto university,kyoto,606-8315, Japan, department of forensic medicine and molecular pathology,school of medicine,kyoto university,kyoto,606-8315, Japan, department of anatomy i,school of medicine,showa university,1-5-8 hatanodai,shinagawa,tokyo,142-8555, Japan
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Authors
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