Advertisement

Combined Voltage and Calcium Imaging and Signal Calibration

  • Marco Canepari
  • Peter Saggau
  • Dejan Zecevic
Chapter

Abstract

Voltage imaging using fluorescent voltage-sensitive dyes can be combined with other optical measurements, in particular with Ca2+ imaging, allowing for correlation of membrane potential changes with intracellular Ca2+ signals. Calibration of fluorescence voltage signals permits the comparison of membrane potential changes from different sites, allowing spatial mapping of membrane potential changes. These two technical aspects enhance the capability of voltage imaging to address several fundamental problems of neurobiology. Here we discuss how to combine voltage imaging with the optical measurement of intracellular Ca2+ transients and different approaches to calibrate voltage-sensitive dye signals on an absolute scale.

Keywords

Optical Signal Calcium Imaging Membrane Potential Change Calcium Indicator Cerebellar Purkinje Neuron 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Antic S, Zecevic D (1995) Optical signals from neurons with internally applied voltage-sensitive dyes. J Neurosci 15:1392–1405.PubMedGoogle Scholar
  2. Beach JM, McGahren ED, Xia J, Duling BR (1996) Ratiometric measurement of endothelial depolarization in arterioles with a potential-sensitive dye. Am J Physiol 270:2216–2227.Google Scholar
  3. Bedlack RS, Wei M-D, Loew LM (1992) Localized membrane depolarizations and localized calcium influx during electric field-guided neurite growth. Neuron 9:393–403.PubMedCrossRefGoogle Scholar
  4. Berger T, Borgdorff A et al. (2007) Combined voltage and calcium epifluorescence imaging in vitro and in vivo reveals subthreshold and suprathreshold dynamics of mouse barrel cortex. J Neurophysiol 97:3751–3762.PubMedCrossRefGoogle Scholar
  5. Bischofberger J, Jonas P (1997) Action potential propagation into the presynaptic dendrites of rat mitral cells. J Physiol (Lond) 504:359–365.CrossRefGoogle Scholar
  6. Brenowitz SD, Regehr WG (2005) Associative short-term synaptic plasticity mediated by endocannabinoids. Neuron 45:419–431.PubMedCrossRefGoogle Scholar
  7. Bullen A, Saggau P (1998) Indicators and optical configuration for simultaneous high-resolution recording of membrane potential and intracellular calcium using laser scanning microscopy. Pflügers Arch 436:788–796.PubMedCrossRefGoogle Scholar
  8. Bullen A, Saggau P (1999) High-speed, random-access fluorescence microscopy: II. Fast quantitative measurements with voltage-sensitive dyes. Biophys J 76:2272–2287.PubMedCrossRefGoogle Scholar
  9. Bullen A, Patel SS, Saggau P (1997) High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators. Biophys J 73:477–491.PubMedCrossRefGoogle Scholar
  10. Canepari M, Vogt KE (2008) Dendritic spike saturation of endogenous calcium buffer and induction of postsynaptic cerebellar LTP. PLoS ONE 3:e4011.PubMedCrossRefGoogle Scholar
  11. Canepari M, Djurisic M, Zecevic D (2007) Dendritic signals from rat hip­pocampal CA1 pyramidal neurons during coincident pre- and post-synaptic activity: a combined voltage- and calcium imaging study. J Physiol 580: 463–484.PubMedCrossRefGoogle Scholar
  12. Canepari M, Vogt K, Zecevic D (2008) Combining voltage and calcium imaging from neuronal dendrites. Cell Mol Neurobiol 58:1079–1093.CrossRefGoogle Scholar
  13. Chen WR, Midtgaard J, Shepherd GM (1997) Forward and backward propagation of dendritic impulses and their synaptic control in mitral cells. Science 278:463–467.PubMedCrossRefGoogle Scholar
  14. Djurisic M, Antic S, Chen WR, Zecevic D (2004) Voltage imaging from dendrites of mitral cells: EPSP attenuation and spike trigger zones. J Neurosci 24:6703–6714.PubMedCrossRefGoogle Scholar
  15. Djurisic M, Popovic M, Carnevale N, Zecevic D (2008) Functional structure of the mitral cell dendritic tuft in the rat olfactory bulb. J Neurosci 28:4057–4068.PubMedCrossRefGoogle Scholar
  16. Eilers J, Konnerth A (2000) Dye loading with patch pipets. In: Yuste R, Lanni F, Konnerth A (eds) Imaging neurons a laboratory manual. Cold Spring Harbour Laboratory Press, New York.Google Scholar
  17. Fierro L, Llano I (1996) High endogenous calcium buffering in Purkinje cells from rat cerebellar slices. J Physiol 496:617–625.PubMedGoogle Scholar
  18. Fluhler E, Burnham VG, Loew LM (1985) Spectra, membrane binding, and potentiometric responses of new charge shift probes. Biochemistry 24:5749–5755.PubMedCrossRefGoogle Scholar
  19. Frick A, Magee J, Johnston D (2004). LTP is accompanied by an enhanced local excitability of pyramidal neuron dendrites. Nat Neurosci 7:126–135.PubMedCrossRefGoogle Scholar
  20. Gross E, Bedlack RS, Loew LM (1994) Dual-wavelength ratiometric fluorescence measurement of the membrane dipole potential. Biophys J 67:208–216.PubMedCrossRefGoogle Scholar
  21. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450.PubMedGoogle Scholar
  22. Gupta RK, Salzberg BM et al. (1981) Improvements in optical methods for ­measuring rapid changes in membrane potential. J Membr Biol 58: 123–137.PubMedCrossRefGoogle Scholar
  23. Hyrc KL, Bownik JM, Goldberg MP (2000). Ionic selectivity of low-affinity ratiometric calcium indicators: mag-Fura-2, Fura-2FF and BTC. Cell Calcium 27:75–86.PubMedCrossRefGoogle Scholar
  24. Kao WY, Davis CE, Kim YI, Beach JM (2001) Fluorescence emission spectral shift measurements of membrane potential in single cells. Biophys J 81:1163–1170.PubMedCrossRefGoogle Scholar
  25. Knisley SB, Justice RK, Kong W, Johnson PL (2000) Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts. Am J Physiol Heart Circ Physiol 279:1421–1433.Google Scholar
  26. Kremer SG, Zeng W, Skorecki KL (1992) Simultaneous fluorescence measurement of calcium and membrane potential responses to endothelin. Am J Physiol 263:1302–1309.Google Scholar
  27. Loew LM, Simpson LL (1981) Charge-shift probes of membrane potential: a probable electrochromic mechanism for p-aminostyrylpyridinium probes on a hemispherical lipid bilayer. Biophys J 34:353–365.PubMedCrossRefGoogle Scholar
  28. Martinez-Zaguilan R, Martinez GM, Lattanzio F, Gillies R J (1991) Simultaneous measurement of intracellular pH and Ca2+ using the fluorescence of SNARF-1 and fura-2. Am J Physiol 260:297–307.Google Scholar
  29. Milojkovic BA, Zhou WL, Antic SD (2007) Voltage and calcium transients in basal dendrites of the rat prefrontal cortex. J Physiol 585:447–468.PubMedCrossRefGoogle Scholar
  30. Montana V, Farkas DL, Loew LM (1989) Dual-wavelength ratiometric fluorescence measurements of membrane potential. Biochemistry 28:4536–4539.PubMedCrossRefGoogle Scholar
  31. Naraghi M (1997) T-jump study of calcium binding kinetics of calcium chelators. Cell Calcium 22:255–268.PubMedCrossRefGoogle Scholar
  32. Neher E (2000) Some quantitative aspects of calcium fluorimetry. In: Yuste R, Lanni F, Konnerth A (eds) Imaging neurons: a laboratory manual. Cold Spring Harbour Laboratory Press, New York.Google Scholar
  33. Roth A, Häusser M (2001) Compartmental models of rat cerebellar Purkinje cells based on simultaneous somatic and dendritic patch-clamp recordings. J Physiol 535:445–472.PubMedCrossRefGoogle Scholar
  34. Sabatini BL, Regehr WG (1996) Timing of neurotransmission at fast synapses in the mammalian brain. Nature 384:170–172.PubMedCrossRefGoogle Scholar
  35. Sabatini BL, Regehr WG (1997) Control of neurotransmitter release by presynaptic waveform at the granule cell to Purkinje cell synapse. J Neurosci 17:3425–3435.PubMedGoogle Scholar
  36. Sabatini BS, Oertner TG, Svoboda K (2002) The life cycle of Ca2+ ions in dendritic spines. Neuron 33:439–452.PubMedCrossRefGoogle Scholar
  37. Schneggenburger R, Meyer AC, Neher E (1999) Released fraction and total size of a pool of immediately available transmitter quanta at a calyx synapse. Neuron 23:399–409.PubMedCrossRefGoogle Scholar
  38. Sinha SR, Saggau P (1999) Simultaneous optical recording of membrane potential and intracellular calcium from brain slices. Methods 18:204–214.PubMedCrossRefGoogle Scholar
  39. Sinha SR, Patel SS, Saggau P (1995) Simultaneous optical recording of evoked and spontaneous transients of membrane potential and intracellular calcium concentration with high spatio-temporal resolution. J Neurosci Methods 60:49–46.PubMedCrossRefGoogle Scholar
  40. Wu JY, Cohen LB (1993) Fast multisite optical measurement of membrane potential. In: Mason WT (ed) Biological techniques: fluorescent and luminescent probes for biological activity. Academic Press, New York.Google Scholar
  41. Yuste R (2000) Loading brain slices with AM esters of calcium indicators. In: Yuste R, Lanni F, Konnerth A (eds) Imaging neurons: a laboratory manual. Cold Spring Harbour Laboratory Press, New York.Google Scholar
  42. Zhang J, Davidson RM, Wei M-D, Loew LM (1998) Membrane electric properties by combined patch clamp and fluorescence ratio imaging in single neurons. Biophys J 74:48–53.PubMedCrossRefGoogle Scholar
  43. Zhou W-L, Yan P, Wuskell JP, Loew LM, Antic SD (2007) Intracellular long-wavelength voltage-sensitive dyes for studying the dynamics of action potentials in axons and thin dendrites. J Neurosci Methods 164:225–239.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Marco Canepari
    • 1
  • Peter Saggau
    • 2
  • Dejan Zecevic
    • 3
  1. 1.Division of Pharmacology and NeurobiologyBiozentrum-University of BaselBaselSwitzerland
  2. 2.Department of NeuroscienceBaylor College of MedicineHoustonUSA
  3. 3.Department of Cellular and Molecular PhysiologyYale University School of MedicineNew HavenUSA

Personalised recommendations