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Using FM4-64 FX to Lable Transport Vesicles of Rat Vascular Smooth Muscle Cells
Jiang Jun1*, Wang Yong2, Li Tao3, Nie Lixia1
1Department of Vascular & Thyroid Surgery, the First Affiliated Hospital of Sichuan Medical University, Luzhou 646000, China; 2The Third Department of Surgery, Xuyong People’s Hospital, Luzhou 646000, China; 3Cardiovascular Institute, Sichuan Medical University, Luzhou 646000, China
Abstract: This study aims to investigate vascular smooth muscle cells (VSMCs) vesicle transport by endocytic vesicles labeling. Rat VSMCs from the thoracic aortic were cultivated in vitro. VSMCs were stimulated with angiotensin II (Ang II) and incubated with FM4-64 FX shortly. Then VSMCs were fixed with paraformaldehyde and marked with angiotensin II type 1 receptor (AT1R) antibody by immunohistochemistry. With Ang II stimulation， VSMCs rapid formed endocytic vesicles and with it， AT1R was transported into the cytoplasm. Presence of an angiotensin receptor blocker (ARB) inhibited the number of endocytic vesicles formation and less AT1R entered the cytoplasm. Macromolecules transport of VSMCs can be illustrated by labeling the intracellular vesicles with FM4-64 FX dye. With this method， early endocytosis of VSMCs when the external environment changed can be investigated.
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