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A naphthyl analog of the aminostyryl pyridinium class of potentiometric membrane dyes shows consistent sensitivity in a variety of tissue, cell, and model membrane preparations

The Journal of Membrane Biology volume 130, pages 1–10 (1992)Cite this article

Summary

The fast potentiometric indicator di-4-ANEPPS is examined in four different preparations: lipid vesicles, red blood cells, squid giant axon, and guinea pig heart. The dye gives consistent potentiometric responses in each of these systems, although some of the detailed behavior varies. In lipid vesicles, the dye displays an increase in fluorescence combined with a red shift of the excitation spectrum upon hyperpolarization. Similar behavior is found in red cells where a dual wavelength ratiometric measurement is also demonstrated. The signal-to-noise ratio of the potentiometric fluorescence response is among the best ever recorded on the voltage-clamped squid axon. The dye is shown to be a faithful and persistent monitor of cardiac action potentials with no appreciable loss of signal or deterioration of cardiac activity for periods as long as 2 hr with intermittent illumination every 10 min. These results, together with previously published applications of the dye to a spherical lipid bilayer model and to cells in culture, demonstrate the versatility of di-4-ANEPPS as a fast indicator of membrane potential.

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Author notes

  1. Eric N. Fluhler

    Present address: Pharmacology Department, RJR-Nabisco, 27102, Winston Salem, North Carolina

Affiliations

  1. Department of Physiology, University of Connecticut Health Center, 06030, Farmington, Connecticut

    Leslie M. Loew & Valerie Montana

  2. Department of Physiology, Yale University School of Medicine, 06510, New Haven, Connecticut

    Lawrence B. Cohen & Wu Jian-young

  3. Marine Biological Laboratory, 02543, Woods Hole, Massachusetts

    Lawrence B. Cohen & Wu Jian-young

  4. Department of Chemistry, State University of New York at Binghamton, 13901, Binghamton, New York

    Leslie M. Loew, James Dix & Eric N. Fluhler

  5. Department of Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania

    Guy Salama

Authors
  1. Leslie M. Loew
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  2. Lawrence B. Cohen
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  3. James Dix
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  4. Eric N. Fluhler
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  5. Valerie Montana
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  6. Guy Salama
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  7. Wu Jian-young
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Additional information

The authors are pleased to acknowledge the support of the U.S. Public Health Service grants GM35063, RR04139 and NS08437. Support from the American Heart Association (871065) and the Whitaker Foundation is also gratefully acknowledged. The detailed comments of a reviewer were most helpful in the preparation of a final version of the manuscript.

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Loew, L.M., Cohen, L.B., Dix, J. et al. A naphthyl analog of the aminostyryl pyridinium class of potentiometric membrane dyes shows consistent sensitivity in a variety of tissue, cell, and model membrane preparations. J. Membarin Biol. 130, 1–10 (1992). https://doi.org/10.1007/BF00233734

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  • DOI: https://doi.org/10.1007/BF00233734

Key Words

  • potential-sensitive dyes
  • red blood cells
  • squid axon
  • fluorescence spectroscopy
  • heart
  • membrane potential