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Gene expression profiling in the human middle cerebral artery after cerebral ischemia
1Department of Experimental Vascular Research, Institution of Medicine, Lund University, Lund, Sweden. petter.vikman@med.lu.se
Abstract:
We have investigated the gene expression in human middle cerebral artery (MCA) after ischemia. Ischemic stroke affects the perfusion in the affected area and experimental cerebral ischemia results in upregulation of vasopressor receptors in the MCA leading to the ischemic area. We obtained human MCA samples distributing to the ischemic area, 7-10 days post-stroke. The gene expression was examined with real-time polymerase chain reaction (PCR) and microarray, proteins were studied with immunohistochemistry. We investigated genes previously shown to be upregulated in animal models of cerebral ischemia (e.g. ET(A), ET(B), AT1, AT2, and 5-HT(2A/1B/1D)). Their mRNA expression was increased compared with controls, consistent with findings in experimental stroke. Immunohistochemistry showed upregulation of the receptors localized on the smooth muscle cells. The gene expression was profiled with microarray and seven genes chosen for further investigation with real-time PCR; ELK3, LY64, Metallothionin IG, POU3F4, Actin alpha2, RhoA and smoothelin. Six of these were regulated the same way when confirming array expression with real-time PCR. Gene expression studies in the human MCA leading to the ischemic region is similar to that seen after MCA occlusion in rats. We found new genes that support the dynamic changes that occur in the MCA distributing to the ischemic region.
Insights
Gene expression in the human middle cerebral artery (MCA) changes after ischemic stroke. Key vasopressor receptors and new genes are upregulated in the MCA supplying the affected brain region.
Area of Science:
- Neuroscience
- Molecular Biology
- Vascular Biology
Background:
- Ischemic stroke impairs blood flow, leading to changes in cerebral arteries.
- Experimental models show altered vasopressor receptor expression in the middle cerebral artery (MCA) after ischemia.
Purpose of the Study:
- To investigate gene expression changes in the human MCA supplying ischemic brain regions.
- To identify specific genes and receptors upregulated post-stroke in the MCA.
Main Methods:
- Human MCA samples were collected 7-10 days after stroke.
- Gene expression analyzed using microarray and real-time PCR.
- Protein localization studied via immunohistochemistry.
Main Results:
- mRNA expression of vasopressor receptors (ET(A), ET(B), AT1, AT2, 5-HT(2A/1B/1D)) was increased in MCA samples from stroke patients.
- Receptor upregulation was confirmed on smooth muscle cells.
- Microarray identified seven candidate genes; six were validated by real-time PCR, including ELK3, LY64, and RhoA.
- Findings in human MCA mirror those in rat models of cerebral ischemia.
Conclusions:
- Human MCA gene expression changes after ischemic stroke are consistent with animal models.
- Upregulation of specific vasopressor receptors and novel genes in the MCA supports dynamic vascular adaptations to ischemia.