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Published on: September 12, 2018
The expression of osteopontin is increased in vessels with blood-brain barrier impairment
1Department of Anaesthesiology and Emergency Medicine, Faculty of Medicine, Kagawa University, Kagawa, Japan.
Insights
Osteopontin expression significantly increases in blood-brain barrier-damaged hippocampal vessels of stroke-prone spontaneously hypertensive rats. This suggests osteopontin plays a role in blood-brain barrier dysfunction in hypertension.
Area of Science:
- Neuroscience
- Cardiovascular Research
- Molecular Biology
Background:
- Stroke-prone spontaneously hypertensive rats (SHRSP) exhibit blood-brain barrier (BBB) dysfunction in hippocampal vessels.
- Investigating gene expression changes in these damaged vessels is crucial for understanding hypertension-related neurological damage.
Purpose of the Study:
- To identify specific genes and proteins altered in BBB-damaged hippocampal vessels of SHRSP.
- To elucidate the role of identified molecules in the pathogenesis of BBB impairment.
Main Methods:
- Microarray analysis of microdissected hippocampal vascular tissue from SHRSP and Wistar-Kyoto (WKY) rats.
- Validation using real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemistry.
Main Results:
- Osteopontin demonstrated the highest gene expression ratio between SHRSP and WKY rats among 8435 genes.
- Osteopontin gene and protein expression were significantly elevated in the hippocampi of SHRSP, particularly in perivascular macrophages/microglial cells.
- Increased osteopontin was localized around hippocampal fissures in SHRSP.
Conclusions:
- Osteopontin expression is upregulated in BBB-damaged hippocampal vessels of hypertensive SHRSP.
- These findings suggest a potential role for osteopontin in the development and progression of blood-brain barrier dysfunction associated with hypertension.
Aims:
We previously reported that the blood-brain barrier (BBB) function was deteriorated in vessels located along hippocampal fissures in stroke-prone spontaneously hypertensive rats (SHRSP). In this study, we examined changes of gene expression in the BBB-damaged vessels of SHRSP.
Methods:
Vascular samples were microdissected from the hippocampi of SHRSP and Wistar-Kyoto (WKY) as a control and the difference in gene expression between the BBB-damaged vessels in SHRSP and vessels without BBB damage in WKY was examined by a microarray. The differences in gene and protein expression between brain tissues in the two strains of rats were examined using real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry.
Results:
The microarray assay revealed that the ratio of osteopontin gene expression in the vascular tissue of the hippocampi of SHRSP to that of WKY was the highest among 8435 genes. Real-time RT-PCR analysis revealed that the gene expression of osteopontin was significantly increased in the hippocampal samples of SHRSP compared with that in the hippocampal samples of WKY rats or with that in the cortical samples of SHRSP. Immunohistochemical and Western blot analyses showed that the osteopontin protein expression was seen in perivascular ED1-positive macrophages/microglial cells located around hippocampal fissures and significantly increased in the hippocampi of SHRSP compared with that of WKY.
Conclusions:
These findings indicate that the expression of osteopontin is increased in BBB-damaged vessels in hypertensive SHRSP compared with that in vessels without BBB impairment in WKY rats, suggesting a role for osteopontin in BBB function.
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