Monitoring Neural Activity and [Ca2+] with Genetically Encoded Ca2+ Indicators

Overview

Journal J Neurosci

Specialty Neurology

Date 2004 Oct 29

PMID 15509744

Citations 113

Authors

Thomas A Pologruto

Ryohei Yasuda

Karel Svoboda

Affiliations

Soon will be listed here.

Abstract

Genetically encoded Ca2+ indicators (GECIs) based on fluorescent proteins (XFPs) and Ca2+-binding proteins [like calmodulin (CaM)] have great potential for the study of subcellular Ca2+ signaling and for monitoring activity in populations of neurons. However, interpreting GECI fluorescence in terms of neural activity and cytoplasmic-free Ca2+ concentration ([Ca2+]) is complicated by the nonlinear interactions between Ca2+ binding and GECI fluorescence. We have characterized GECIs in pyramidal neurons in cultured hippocampal brain slices, focusing on indicators based on circularly permuted XFPs [GCaMP (Nakai et al., 2001), Camgaroo2 (Griesbeck et al., 2001), and Inverse Pericam (Nagai et al., 2001)]. Measurements of fluorescence changes evoked by trains of action potentials revealed that GECIs have little sensitivity at low action potential frequencies compared with synthetic [Ca2+] indicators with similar affinities for Ca2+. The sensitivity of GECIs improved for high-frequency trains of action potentials, indicating that GECIs are supralinear indicators of neural activity. Simultaneous measurement of GECI fluorescence and [Ca2+] revealed supralinear relationships. We compared GECI fluorescence saturation with CaM Ca2+-dependent structural transitions. Our data suggest that GCaMP and Camgaroo2 report CaM structural transitions in the presence and absence of CaM-binding peptide, respectively.

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References

Miyawaki A, Griesbeck O, HEIM R, Tsien R . Dynamic and quantitative Ca2+ measurements using improved cameleons. Proc Natl Acad Sci U S A. 1999; 96(5):2135-40. PMC: 26749. DOI: 10.1073/pnas.96.5.2135. View

Mirzoeva S, Weigand S, Lukas T, Shuvalova L, Anderson W, Watterson D . Analysis of the functional coupling between calmodulin's calcium binding and peptide recognition properties. Biochemistry. 1999; 38(13):3936-47. DOI: 10.1021/bi9821263. View

Smetters D, Majewska A, Yuste R . Detecting action potentials in neuronal populations with calcium imaging. Methods. 1999; 18(2):215-21. DOI: 10.1006/meth.1999.0774. View

Mainen Z, Maletic-Savatic M, Shi S, Hayashi Y, Malinow R, Svoboda K . Two-photon imaging in living brain slices. Methods. 1999; 18(2):231-9, 181. DOI: 10.1006/meth.1999.0776. View

Sun H, Yin D, Squier T . Calcium-dependent structural coupling between opposing globular domains of calmodulin involves the central helix. Biochemistry. 1999; 38(38):12266-79. DOI: 10.1021/bi9818671. View

Baird G, Zacharias D, Tsien R . Circular permutation and receptor insertion within green fluorescent proteins. Proc Natl Acad Sci U S A. 1999; 96(20):11241-6. PMC: 18018. DOI: 10.1073/pnas.96.20.11241. View

Maravall M, Mainen Z, Sabatini B, Svoboda K . Estimating intracellular calcium concentrations and buffering without wavelength ratioing. Biophys J. 2000; 78(5):2655-67. PMC: 1300854. DOI: 10.1016/S0006-3495(00)76809-3. View

McAllister A, Stevens C . Nonsaturation of AMPA and NMDA receptors at hippocampal synapses. Proc Natl Acad Sci U S A. 2000; 97(11):6173-8. PMC: 18577. DOI: 10.1073/pnas.100126497. View

Kerr R, Lev-Ram V, Baird G, Vincent P, Tsien R, Schafer W . Optical imaging of calcium transients in neurons and pharyngeal muscle of C. elegans. Neuron. 2000; 26(3):583-94. DOI: 10.1016/s0896-6273(00)81196-4. View

10.

Sabatini B, Svoboda K . Analysis of calcium channels in single spines using optical fluctuation analysis. Nature. 2000; 408(6812):589-93. DOI: 10.1038/35046076. View

11.

Nakai J, Ohkura M, Imoto K . A high signal-to-noise Ca(2+) probe composed of a single green fluorescent protein. Nat Biotechnol. 2001; 19(2):137-41. DOI: 10.1038/84397. View

12.

Nagai T, Sawano A, Park E, Miyawaki A . Circularly permuted green fluorescent proteins engineered to sense Ca2+. Proc Natl Acad Sci U S A. 2001; 98(6):3197-202. PMC: 30630. DOI: 10.1073/pnas.051636098. View

13.

Griesbeck O, Baird G, Campbell R, Zacharias D, Tsien R . Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications. J Biol Chem. 2001; 276(31):29188-94. DOI: 10.1074/jbc.M102815200. View

14.

Robert V, Gurlini P, Tosello V, Nagai T, Miyawaki A, Di Lisa F . Beat-to-beat oscillations of mitochondrial [Ca2+] in cardiac cells. EMBO J. 2001; 20(17):4998-5007. PMC: 125611. DOI: 10.1093/emboj/20.17.4998. View

15.

Saimi Y, Kung C . Calmodulin as an ion channel subunit. Annu Rev Physiol. 2002; 64:289-311. DOI: 10.1146/annurev.physiol.64.100301.111649. View

16.

Sabatini B, Oertner T, Svoboda K . The life cycle of Ca(2+) ions in dendritic spines. Neuron. 2002; 33(3):439-52. DOI: 10.1016/s0896-6273(02)00573-1. View

17.

Star E, Kwiatkowski D, Murthy V . Rapid turnover of actin in dendritic spines and its regulation by activity. Nat Neurosci. 2002; 5(3):239-46. DOI: 10.1038/nn811. View

18.

Lisman J, Schulman H, Cline H . The molecular basis of CaMKII function in synaptic and behavioural memory. Nat Rev Neurosci. 2002; 3(3):175-90. DOI: 10.1038/nrn753. View

19.

Oertner T, Sabatini B, Nimchinsky E, Svoboda K . Facilitation at single synapses probed with optical quantal analysis. Nat Neurosci. 2002; 5(7):657-64. DOI: 10.1038/nn867. View

20.

Komeiji Y, Ueno Y, Uebayasi M . Molecular dynamics simulations revealed Ca(2+)-dependent conformational change of Calmodulin. FEBS Lett. 2002; 521(1-3):133-9. DOI: 10.1016/s0014-5793(02)02853-3. View