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http://dx.doi.org/10.3340/jkns.2014.55.6.307

Semi-Quantitative Analyses of Hippocampal Heat Shock Protein-70 Expression Based on the Duration of Ischemia and the Volume of Cerebral Infarction in Mice  

Choi, Jong-Il (Department of Neurosurgery, Ansan Hospital, Korea University Medical Center, Korea University College of Medicine)
Kim, Sang-Dae (Department of Neurosurgery, Ansan Hospital, Korea University Medical Center, Korea University College of Medicine)
Kim, Se-Hoon (Department of Neurosurgery, Ansan Hospital, Korea University Medical Center, Korea University College of Medicine)
Lim, Dong-Jun (Department of Neurosurgery, Ansan Hospital, Korea University Medical Center, Korea University College of Medicine)
Ha, Sung-Kon (Department of Neurosurgery, Ansan Hospital, Korea University Medical Center, Korea University College of Medicine)
Publication Information
Journal of Korean Neurosurgical Society / v.55, no.6, 2014 , pp. 307-312 More about this Journal
Abstract
Objective : We investigated the expression of hippocampal heat shock protein 70 (HSP-70) infarction volume after different durations of experimental ischemic stroke in mice. Methods : Focal cerebral ischemia was induced in mice by occluding the middle cerebral artery with the modified intraluminal filament technique. Twenty-four hours after ischemia induction, both hippocampi were extracted for HSP-70 protein analyses. Slices from each hemisphere were stained with 2,3,5-triphenyltetrazolium chloride (2%), and infarction volumes were calculated. HSP-70 levels were evaluated using western blot and enzyme-linked immunosorbent assay (ELISA). HSP-70 subtype (hsp70.1, hspa1a, hspa1b) mRNA levels in the hippocampus were measured using reverse transcription-polymerase chain reaction (RT-PCR). Results : Cerebral infarctions were found ipsilateral to the occlusion in 10 mice exposed to transient ischemia (5 each in the 30-min and 60-min occlusion groups), whereas no focal infarctions were noted in any of the sham mice. The average infarct volumes of the 2 ischemic groups were $22.28{\pm}7.31mm^3$ [30-min group${\times}$standard deviation (SD)] and $38.06{\pm}9.53mm^3$ (60-min group${\times}$SD). Western blot analyses and ELISA showed that HSP-70 in hippocampal tissues increased in the infarction groups than in the sham group. However, differences in HSP-70 levels between the 2 infarction groups were statistically insignificant. Moreover, RT-PCR results demonstrated no relationship between the mRNA expression of HSP-70 subtypes and occlusion time or infarction volume. Conclusion : Our results indicated no significant difference in HSP-70 expression between the 30- and 60-min occlusion groups despite the statistical difference in infarction volumes. Furthermore, HSP-70 subtype mRNA expression was independent of both occlusion duration and cerebral infarction volume.
Keywords
Cerebral infarction volume; Heat shock protein; Occlusion time;
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