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Fluorescent styryl dyes applied as fast optical probes of cardiac action potential

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Abstract

Several styryl dyes were tested as fast optical probes of membrane action potentials in mammalian heart muscle tissue. After staining, atrial specimens were superfused in physiological salt solution, and fluorescence was excited by an argon ion laser. Excitation spot size on the surface of the preparation was 60 μm in diameter. Dyes RH 160, RH 237, and RH 421 performed excellently as fast fluorescent probes of cardiac membrane potential. Fractional fluorescence changes, ΔF/F, due to the action potential were in the range 2 to 6% at 514.5 nm excitation. Rise times of the action potential onset detected with each of the dyes were less than 0.5 ms, which is as fast or even faster than microelectrode measurements (atria of the rat). Thus membrane potential changes could be monitored with high resolution in both time and space. Emission spectra from heart muscle preparations stained with these dyes were shifted to shorter wavelengths by 70 nm and more as compared to spectra of the dyes in ethanol solution. The fluorescence spectrum of RH 160 at resting potential and the spectrum recorded during the plateau phases of the action potential were measured and showed no difference within the spectral resolution. As can be concluded from measurements of fluorescence changes at different excitation wavelengths, electrochromism cannot be the only mechanism causing the potential response.

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References

  • Cleemann L, Pizzaro G, Morad M (1984) Optical measurements of extracellular calcium depletion during a single heart beat. Science 226:174–177

    Google Scholar 

  • Cohen LB, Salzberg BM (1978) Optical measurement of membrane potential. Rev Physiol Biochem Pharmacol 83:35–88

    Google Scholar 

  • Conti F (1975) Fluorescent probes in nerve membranes. Annu Rev Biophys Bioeng 4:287–310

    Google Scholar 

  • Dillon S, Morad M (1981) A new laser scanning system for measuring action potential propagation in the heart. Science 214:453–456

    Google Scholar 

  • Fluhler E, Burnham VG, Loew LM (1985) Spectra, membrane binding, and potentiometric response of new charge shift probes. Biochemistry 24:5749–5755

    Google Scholar 

  • Fujii S, Hirota A, Kamino K (1981a) Optical recording of development of electrical activity in embryonic chick heart during early phases of cardiogenesis. J Physiol 311:147–160

    Google Scholar 

  • Fujii S, Hirota A, Kamino K (1981b) Optical recording of pacemaker potential and rhythm generation in early embryonic chick heart. J Physiol 312:253–263

    Google Scholar 

  • Grinvald A (1984) Real-time optical imaging of neuronal activity. Trends Neurosci 7:143–150

    Google Scholar 

  • Grinvald A, Manker A, Segal M (1981) Visualization of the spread of electrical activity in rat hippocamal slices by voltage-sensitive optical probes. J Physiol 333:269–291

    Google Scholar 

  • Grinvald A, Hildesheim R, Farber IC, Anglister L (1982) Improved fluorescent probes for the measurement of rapid changes in membrane potential. Biophys J 39:301–308

    Google Scholar 

  • Grinvald A, Fine A, Farber IC, Hildesheim R (1983) Fluorescence monitoring of electrical responses from small neurons and their processes. Biophys J 42:195–198

    Google Scholar 

  • Grinvald A, Anglister L, Freeman JA, Hildesheim R, Manker A (1984) Real-time optical imaging of naturally evoked electrical activity in intact frog brain. Nature 308:848–850

    Google Scholar 

  • Hill BC, Courtney KR (1982) Voltage-sensitive dyes. Discerning contraction and electrical signals in myocardium. Biophys J 40:255–257

    Google Scholar 

  • Hirota A, Fujii S, Kamino K (1979) Optical monitoring of spontaneous electrical activity of 8-somite embryonic chick heart. Jpn J Physiol 29:635–639

    Google Scholar 

  • Kagawa Y, Racker E (1971) Partial resolution of the enzymes catalyzing oxidative phosphorylation. J Biol Chem 246: 5477–5487

    Google Scholar 

  • Kamino K, Hirota A, Fujii S (1981) Localization of pacemaking activity in early embryonic heart monitored using voltage-sensitive dye. Nature 290:595–597

    Google Scholar 

  • Krasne S (1980a) Interactions of voltage-sensitive dyes with membranes. I. Steady-state permeability behaviors induced by cyanine dyes. Biophys J 30:415–439

    Google Scholar 

  • Krasne S (1980b) Interactions of voltage-sensitive dyes with membranes. II. Spectrophotometric and electrical correlates of cyanine-dye adsorption to membranes. Biophys J 30:441–462

    Google Scholar 

  • Loew LM (1982) Design and characterization of electrochromic membrane probes. J Biochem Biophys Methods 6:243–260

    Google Scholar 

  • Loew LM, Bonneville GW, Surow J (1978) Charge shift optical probes of membrane potential. Theory Biochemistry 17:4065–4071

    Google Scholar 

  • Loew LM, Scully S, Simpson L, Waggoner AS (1979) Evidence for a charge-shift electrochromic mechanism in a probe of membrane potential. Nature 281:497–499

    Google Scholar 

  • Loew LM, Cohen LB, Salzberg BM, Obaid AL, Bezanilla F (1985) Charge shift probes of membrane potential. Characterization of Aminostyrylpyridinium dyes on the squid giant axon. Biophys J 47:71–77

    Google Scholar 

  • Morad M, Salama G (1979) Optical probes of membrane potential in heart muscle. J Physiol 292:267–295

    Google Scholar 

  • Platt JR (1961) Electrochromism, a possible change of color producible in dyes by an electric field. J Chem Phys 34:862–863

    Google Scholar 

  • Ross WN, Salzberg BM, Cohen LB, Davila HV (1974) A large change in dye absorption during the action potential. Biophys J 14:983–986

    Google Scholar 

  • Sakai T, Fujii S, Hirota A, Kamino K (1983) Optical evidence for calcium-action potentials in early embryonic precontractile chick heart using a potential-sensitive dye. J Membr Biol 72:205–212

    Google Scholar 

  • Salama G, Morad M (1976a) Optical measurements of cardiac action potential with merocyanine 540. Abstract F-AM-A8. Biophys J 16:153a

    Google Scholar 

  • Salama G, Morad M (1976b) Merocyanine 540 as an optical probe of transmembrane electrical activity in the heart. Science 191:485–487

    Google Scholar 

  • Sawanobori T, Hirano Y, Hirota A, Fujii S (1984) Circusmovement tachycardia in frog atrium monitored by voltage-sensitive dyes. Am J Physiol 247 (Heart Circ Physiol 16): H185-H194

    Google Scholar 

  • Schindler HG, Quast U (1980) Functional acetylcholine receptor from Torpedeo marmaorata in planar membranes. Proc Natl Acad Sci USA 77:3052–3056

    Google Scholar 

  • Tasaki I, Warashina A, Pant H (1975) Studies of light emission, absorption and energy transfer in nerve membranes labelled with fluorescent probes. Biophys Chem 4:1–13

    Google Scholar 

  • Tasaki I, Warashina A (1976) Dye-membrane interaction and its changes during nerve excitation. Photochem Photobiol 24:191–207

    Google Scholar 

  • Tritthart H, MacLeod DP, Stierle HE, Krause H (1973) Effects of Ca-free and EDTA-containing tyrode solution on transmembrane electrical activity and contraction in guinea-pig papillary muscle. Pflügers Arch 338:361–376

    Google Scholar 

  • Waggoner AS (1976) Optical probes of membrane potential. J Membr Biol 27:317–334

    Google Scholar 

  • Waggoner AS (1979) Dye indicators of membrane potential. Annu Rev Biophys Bioeng 8:47–68

    Google Scholar 

  • Waggoner AS, Grinvald A (1977) Mechanisms of rapid optical changes of potential sensitive dyes. Ann NY Acad Sci 303:217–241

    Google Scholar 

  • Waggoner AS, Sirkin D, Tolles R, Wang CH (1975) Rate of membrane penetration of potential-sensitive dyes. Biophys J 15:20a (Abstr)

    Google Scholar 

  • Waggoner AS, Wang CH, Tolles RL (1977) Mechanism of potential-dependent light absorption changes of lipid bilayer membranes in the presence of cyanine and oxonol dyes. J Membr Biol 33:109–140

    Google Scholar 

  • Warashina A, Tasaki I (1975) Evidence for rotation of dye molecules in membrane macromolecules associated with nerve excitation. Proc Jpn Acad 51:610–615

    Google Scholar 

  • Windisch H, Müller W (1983) Cardiac effects of voltage-sensitive dyes for photometric measurements of cardiac action potentials. Naunyn Schmiedebergs Arch Pharmacol 322:R25 (Abstr)

    Google Scholar 

  • Windisch H, Tritthart HA (1981) Calcium ion effects on the rising phases of action potentials obtained from guinea-pig papillary muscles at different potassium concentrations. J Mol Cell Cardiol 13:457–469

    Google Scholar 

  • Windisch H, Müller W, Tritthart HA (1985) Fluorescence monitoring of rapid changes in membrane potential in heart muscle. Biophys J 48:877–884

    Google Scholar 

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Müller, W., Windisch, H. & Tritthart, H.A. Fluorescent styryl dyes applied as fast optical probes of cardiac action potential. Eur Biophys J 14, 103–111 (1986). https://doi.org/10.1007/BF00263067

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  • DOI: https://doi.org/10.1007/BF00263067

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