Abstract
The use of single- and dual-wavelength Ca2+-sensitive fluorescent dyes to monitor changes in endothelial and/or smooth muscle intracellular Ca2+ levels has provided information linking Ca2+ events to changes in arterial function. Here we describe the in vitro techniques used to selectively load Ca2+ indicators into either the endothelium or the smooth muscle of cannulated rat cremaster arteries. These vessels normally develop spontaneous myogenic tone that is largely unaffected by the loading of Ca2+ indicators or the subsequent imaging procedures. This suggests that there is minimal Ca2+ buffering or damage, and that the fluorescent indicator-loaded vessels behave similarly to unloaded preparations. Importantly, these approaches are applicable to both isobaric and isometric preparations and have been also used for the study of a number of vascular beds including cerebral, mesenteric, coronary, and skeletal muscle vasculatures.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McSherry IN, Sandow SL, Campbell WB, Falck JR, Hill MA, Dora KA (2006) A role for heterocellular coupling and EETs in dilation of rat cremaster arteries. Microcirculation 13:119–130
Kansui Y, Garland CJ, Dora KA (2008) Enhanced spontaneous Ca2+ events in endothelial cells reflects signalling through myoendothelial gap junctions in pressurized mesenteric arteries. Cell Calcium 44:135–146
Ledoux J, Taylor MS, Bonev AD, Hannah RM, Solodushko V, Shui B, Tallini Y, Kotlikoff MI, Nelson MT (2008) Functional architecture of inositol 1,4,5-trisphosphate signaling in restricted spaces of myoendothelial projections. Proc Natl Acad Sci U S A 105:9627–9632
Tallini Y, Brekke J, Shui B, Doran R, Hwang SM, Nakai J, Salama G, Segal S, Kotlikoff M (2007) Propagated endothelial Ca2+ waves and arteriolar dilation in vivo. Measurements in Cx40BAC GCaMP2 transgenic mice. Circ Res 101:1300–1309
Meininger GA, Zawieja DC, Falcone JC, Hill MA, Davey JP (1991) Calcium measurement in isolated arterioles during myogenic and agonist stimulation. Am J Physiol Heart Circ Physiol 261:H950–H959
Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450
Hyrc KL, Rzeszotnik Z, Kennedy BR, Goldberg MP (2007) Determining calcium concentration in heterogeneous model systems using multiple indicators. Cell Calcium 42:576–589
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Dora, K.A., Hill, M.A. (2013). Measurement of Changes in Endothelial and Smooth Muscle Ca2+ in Pressurized Arteries. In: Lambert, D., Rainbow, R. (eds) Calcium Signaling Protocols. Methods in Molecular Biology, vol 937. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-086-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-62703-086-1_14
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-085-4
Online ISBN: 978-1-62703-086-1
eBook Packages: Springer Protocols