Semi-Quantitative Analyses of Hippocampal Heat Shock Protein-70 Expression Based on the Duration of Ischemia and the Volume of Cerebral Infarction in Mice.
10.3340/jkns.2014.55.6.307
- Author:
Jong Il CHOI
1
;
Sang Dae KIM
;
Se Hoon KIM
;
Dong Jun LIM
;
Sung Kon HA
Author Information
1. Department of Neurosurgery, Ansan Hospital, Korea University Medical Center, Korea University College of Medicine, Ansan, Korea. sungkha@yahoo.com
- Publication Type:Original Article
- Keywords:
Cerebral infarction volume;
Heat shock protein;
Occlusion time
- MeSH:
Animals;
Blotting, Western;
Brain Ischemia;
Cerebral Infarction*;
Enzyme-Linked Immunosorbent Assay;
Heat-Shock Proteins;
Hippocampus;
Hot Temperature*;
HSP70 Heat-Shock Proteins;
Infarction;
Ischemia*;
Mice*;
Middle Cerebral Artery;
RNA, Messenger;
Shock*;
Stroke
- From:Journal of Korean Neurosurgical Society
2014;55(6):307-312
- CountryRepublic of Korea
- Language:English
-
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+/-7.31 mm3 [30-min group+/-standard deviation (SD)] and 38.06+/-9.53 mm3 (60-min group+/-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.
