Isolation of RNA from Laser-Microdissected Tissue for Comparison of KLF Transcription in Intra- versus Extra-cranial Vascular Smooth Muscle
Gustafson, Tyler Joseph
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Large intracranial arteries are unique compared to conduit vessels in the extracranial vasculature. They differ in resistance, blood flow regulation, reactivity to vasostimuli, and the accumulation of atherosclerotic plaque formation. Large cerebral vessels are atheroprotective, while conduits are atheroprone. Central to the observed differences are the vascular smooth muscle cells (VSMCs). Unique transcriptional activity in cerebral VSMCs may be responsible for the atheroprotective nature of the intracranial vasculature. Krüppel-Like Factor (KLF) 4 helps promote cellular de-differentiation, while KLF5 facilitates proliferation. Working together, the two KLFs may facilitate the early onset of atherosclerosis in the aorta, relative to the cerebral vasculature. The current study aimed to address whether or not those KLFs were partially responsible for the observed differences in the two types of arteries. Laser microdissection was used to isolate VSMCs from the aorta and basilar artery to measure KLF abundance using rt-qPCR.