Focal Cerebral Ischemia Induces Decrease of Astrocytic Phosphoprotein PEA-15 in Brain Tissue and HT22 Cells

Phil-Ok Koh

doi:10.5625/lar.2010.26.3.311

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Koh: Focal Cerebral Ischemia Induces Decrease of Astrocytic Phosphoprotein PEA-15 in Brain Tissue and HT22 Cells

Original Article

Lab. Anim. Res. 2010;26(3):311-314.

Published online: 17 September 2010

DOI: https://doi.org/10.5625/lar.2010.26.3.311

Focal Cerebral Ischemia Induces Decrease of Astrocytic Phosphoprotein PEA-15 in Brain Tissue and HT22 Cells

Phil-Ok Koh^∗

Department of Anatomy, College of Veterinary Medicine and Research Institute of Life Sciences, Gyeongsang National University, Jinju, Korea

^∗Corresponding author: Phil Ok Koh, Department of Anatomy, College of Veterinary Medicine, Gyeongsang National University, 900 Gajwa-dong, Jinju, Gyeongnam 660-701, Korea Tel: +82-55-751-5809 Fax: +82-55-751-5803 E-mail: pokoh@gnu.ac.kr

Received 13 July 2010 Revised 31 July 2010 Accepted 7 September 2010

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

PEA-15 is a small phosphoprotein (15 kDa) that is enriched in brain astrocytes. PEA-15 acts as an important modulator of cellular function including apoptosis and signal integration. This study investigated the expression of PEA-15 in focal cerebral ischemic injury. Cerebral ischemia was surgically induced in adult male rats by middle cerebral artery occlusion (MCAO), and brains were collected 24 hr after MCAO. A proteomic approach demonstrated decreases of PEA-15 protein spots in MCAO-operated animals in comparison to sham-operated animals. Western blot analysis clearly demonstrated that MCAO induces decreases in PEA-15 levels. We previously showed that glutamate toxicity induces cell death in a hippocampus-derived cell line (HT22). Glutamate exposure induces decreases of PEA-15 levels in HT22 cells. The results of this study suggest that focal cerebral ischemia induces cell death through downregulation of PEA-15 protein.

Keywords: HT22 cells, middle cerebral artery occlusion, PEA-15

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Figure 1.

PEA-15 protein spots identified by MALDI-TOF. Arrows indicate the protein spots. The intensity of spots was measured using PDQuest software. The ratio of intensity is described as spots intensity of MCAO-operated animal to spots intensity of sham-operated animal. Data are shown as mean± S.E.M. ∗P<0.05 (vs. Sham). Mw and IP indicate molecular weight and isoelectrical point, respectively.

Figure 2.

Western blot analysis of PEA-15 in the cerebral cortex from sham-operated and MCAO-operated animals. Each lane represents an individual experimental animal. Densitometric analysis of PEA-15 levels is represented as intensity of PEA-15 to intensity of actin. Data are shown as mean±S.E.M. ∗P<0.05 (vs. Sham).

Figure 3.

Western blot analysis of PEA-15 in HT22 cells. Vehicle or glutamate (5 mM) was exposed to HT22 cells for 24 hr. Each lane represents an individual experimental animal. Densitometric analysis of PEA-15 levels is represented as intensity of PEA-15 to intensity of actin. Data are shown as mean±S.E.M. ∗P<0.05 (vs. Vehicle).

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