Abstract
Changes in intracellular free calcium ion concentration ([Ca2+]i) play a major role in vascular smooth muscle cell function (1,2). Elevation of [Ca2+]i is an important regulator of multiple downstream signaling pathways and it is a major determinant of vascular smooth muscle contraction (1–3). Agonists, such as angiotensin II (Ang II), endothelin-1, and vasopressin, that mediate effects through G protein-coupled receptors, increase [Ca2+]i by inducing Ca2+mobilization from intracellular sarcoplasmic/endoplasmic reticular stores, and by stimulating transplasmalemmal Ca2+influx (4–6). One of the earliest measurable events resulting from Ang II stimulation of vascular smooth muscle cells, is a rapid, phospholipase C (PLC)-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate, to produce two second messengers, 1,2 diacylglycerol and inositol 1,4,5-trisphosphate (IP3) (7,8). The water-soluble messenger IP3, binds to specific IP3 receptors to release Ca2+from nonmitochondrial intracellular stores.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Claphan, D. E. (1995) Calcium signaling. Cell 80, 259–268.
Berridge, M. J. and Dupont, G. (1994) Spatial and temporal signaling by calcium. Curr. Opin. Cell Biol. 6, 267–274.
Karaki, H., Ozaki, H., Hori, M., Mitsui-Saito, M., Amano, K., Harada, K., et al. (1997) Calcium movements, distribution and functions in smooth muscle. Pharmacol. Rev. 49, 157–230.
Touyz, R. M. and Schiffrin, E. L. (1997) Role of calcium influx and intracellular calcium stores in angiotensin II-mediated calcium hyperresponsiveness in smooth muscle from spontaneously hypertensive rats. J. Hypertens. 15, 1431–1439.
Touyz, R. M., Deng, L-Y., Li, J-S., and Schiffrin, E. L. (1996) Differential effects of vasopressin and endothelin-1 on vascular contractile and calcium responses in pressurized small arteries from spontaneously hypertensive rats. J. Hypertens. 14, 983–991.
Douglas, S. A. and Ohlstein, E. H. (1997) Signal transduction mechanisms mediating the vascular actions of endothelin. J. Vasc. Res. 34, 152–164
Berridge, M. J. (1993) Inositol trisphosphate and calcium signaling. Nature 361, 315–325.
Griendling, K. K., Ushio-Fukai, M., Lassègue, B., and Alexander, R. W. (1997) Angiotensin II signaling in vascular smooth muscle. New concepts. Hypertension 29, 366–373.
Ridgway, E. B. and Ashley, C. C. (1967) Calcium transients in single muscle fibres. Biochem. Biophys. Res. Commun. 29, 229–234.
Murphey, E., Coll, K., Rich, T. L., and Williamson, J. R. (1980) Hormonal effects on calcium homeostasis in isolated hepatocytes. J. Biol. Chem. 255, 6600–6608.
Tsien, R. W. (1981) A non-disruptive technique for loading buffers and indicators into cells. Nature 290, 527–528.
Tsien, R. Y., Pozzan, T., and Rink, T. J.(1982) Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J. Cell Biol. 94, 325–334.
Roe, M. W., Lemasters, J. J., and Herman, B. (1990) Assessment of fura-2 for measurements of cytosolic free calcium. Cell Calcium 11, 63–73.
Tsien, R. Y., Rink, T. J., and Poenie, M. (1985) Measurement of cytosolic free Ca2+in individual cells using fluorescence microscopy with dual excitation wavelengths. Cell Calcium 6, 145–157.
Williams, D. A. (1995) Fluorescence imaging of cytosolic calcium, in Measurement and Manipulation of Intracellular Ions, vol. 27 (Kraicer, J. and Dixon, S. J., eds.), Academic, New York, pp. 69–81.
Goldman, W. F., Bova, S., and Blaustein, M. P. (1990) Measurement of intracellular Ca2+in cultured arterial smooth muscle cells using fura-2 and digital imaging microscopy. Cell Calcium 11, 221–231.
Grynkiewicz, G., Poenie, M., and Tsien, R. Y. (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biochem. Sci. 260, 3440–3450.
Williams, D. A. and Fay, F. S. (1990) Intracellular calibration of the fluorescent calcium indicator fura-2. Cell Calcium 11, 75–63.
Iredale, P. A. and Dickenson, J. M. (1995) Measurement of intracellular free calcium ion concentration in cell populations using fura-2, in Signal Transduction Protocols, vol. 41 (Kendall, D. A. and Hill, S. J., eds.), Humana, Totowa, NJ, pp. 203–213.
Moore, E. D. W., Becker, P. L., Fogarty, K. E., Williams, D. A., and Fay, F. S. (1990) Ca2+imaging in single living cells: theoretical and practical issues. Cell Calcium 11, 157–179.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Touyz, R.M., Schiffrin, E.L. (2001). Measurement of Intracellular Free Calcium Ion Concentration in Vascular Smooth Muscle Cells. In: Wang, D.H. (eds) Angiotensin Protocols. Methods in Molecular Medicine™, vol 51. Humana Press. https://doi.org/10.1385/1-59259-087-X:341
Download citation
DOI: https://doi.org/10.1385/1-59259-087-X:341
Publisher Name: Humana Press
Print ISBN: 978-0-89603-763-2
Online ISBN: 978-1-59259-087-2
eBook Packages: Springer Protocols