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Heterogeneity of [Ca²⁺]_i signaling in intact rat aortic endothelium

Department of Physiology, National Cheng-Kung University Medical College, Tainan 701, Taiwan

¹Correspondence: Department of Physiology, College of Medicine, National Cheng-Kung University Medical College, #1, Ta-Hsiue Rd., Tainan 701, Taiwan. E-mail: jen{at}mail.ncku.edu.tw

Most existing knowledge about [Ca²⁺]_i signaling in vascular endothelium has been based on studies using endothelial cells cultured in vitro. To examine how endothelial cells behave in situ, we have developed a method to monitor single-cell [Ca²⁺]_i from Fura-2-loaded rat aortic segments. Fluorescence ratio images from large numbers of endothelial cells were acquired by using a flow chamber mounted on a dual-wavelength fluorescence microscope. Our results showed that either acetylcholine or histamine reversibly activated the vascular endothelium by eliciting M₃ or H₁ receptor-mediated [Ca²⁺]_i increases, respectively. The acetylcholine-evoked endothelial [Ca²⁺]_i elevation at the branch site (intercostal orifice) was much more pronounced than that at the non-branch area. However, endothelium at the branch site was relatively insensitive to histamine. Both acetylcholine-sensitive and histamine-sensitive endothelial cells were arranged in belts aligned along flow lines and were intercalated with each other. Data analyzed from 400 endothelial cells located at the non-branch site showed drastically heterogeneous [Ca²⁺]_i responses to a fixed concentration of either acetylcholine or histamine, differing by two orders of magnitude in individual cells. As a conclusion, vascular endothelial cells appear to have their own characteristic [Ca²⁺]_i ‘fingerprint’ to various agonists and they may function coordinately in situ.—Huang, T.-Y., Chu, T.-F., Chen, H.-i., Jen, C. J. Heterogeneity of [Ca²⁺]_i signaling in intact rat aortic endothelium.

Key Words: atherosclerosis • acetylcholine • histamine • blood vessel • endothelial cells

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