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Evaluation of Voltage-Sensitive Fluorescence Dyes for Monitoring Neuronal Activity in the Embryonic Central Nervous System

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Abstract

Using an optical imaging technique with voltage-sensitive dyes (VSDs), we investigated the functional organization and architecture of the central nervous system (CNS) during embryogenesis. In the embryonic nervous system, a merocyanine-rhodanine dye, NK2761, has proved to be the most useful absorption dye for detecting neuronal activity because of its high signal-to-noise ratio (S/N), low toxicity and small dye bleaching. In the present study, we evaluated the suitability of fluorescence VSDs for optical recording in the embryonic CNS. We screened eight styryl (hemicyanine) dyes in isolated brainstem–spinal cord preparations from 7-day-old chick embryos. Measurements of voltage-related optical signals were made using a multiple-site optical recording system. The signal size, S/N, photobleaching, effects of perfusion and recovery of neural responses after staining were compared. We also evaluated optical responses with various magnifications. Although the S/N was lower than with the absorption dye, clear optical responses were detected with several fluorescence dyes, including di-2-ANEPEQ, di-4-ANEPPS, di-3-ANEPPDHQ, di-4-AN(F)EPPTEA, di-2-AN(F)EPPTEA and di-2-ANEPPTEA. Di-2-ANEPEQ showed the largest S/N, whereas its photobleaching was faster and the recovery of neural responses after staining was slower. Di-4-ANEPPS and di-3-ANEPPDHQ also exhibited a large S/N but required a relatively long time for recovery of neural activity. Di-4-AN(F)EPPTEA, di-2-AN(F)EPPTEA and di-2-ANEPPTEA showed smaller S/Ns than di-2-ANEPEQ, di-4-ANEPPS and di-3-ANEPPDHQ; but the recovery of neural responses after staining was faster. This study demonstrates the potential utility of these styryl dyes in optical monitoring of voltage changes in the embryonic CNS.

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References

  • Acker CD, Yan P, Loew LM (2011) Single-voxel recording of voltage transients in dendritic spines. Biophys J 101:L11–L13

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Baker BJ, Kosmidis EK, Vucinic D, Falk CX, Cohen LB, Djurisic M, Zecevic D (2005) Imaging brain activity with voltage- and calcium-sensitive dyes. Cell Mol Neurobiol 25:245–282

    Article  CAS  PubMed  Google Scholar 

  • Canepari M, Zecevic D (2010) Membrane potential imaging in the nervous system. Springer, New York

    Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Cohen LB, Salzberg BM, Davila HV, Ross WN, Landowne D, Waggoner AS, Wang CH (1974) Changes in axon fluorescence during activity: molecular probes of membrane potential. J Membr Biol 19:1–36

    Article  CAS  PubMed  Google Scholar 

  • Ebner TJ, Chen G (1995) Use of voltage-sensitive dyes and optical recordings in the central nervous system. Prog Neurobiol 46:463–506

    Article  CAS  PubMed  Google Scholar 

  • Fisher JAN, Barchi JR, Welle CG, Kim G-H, Kosterin P, Obaid AL, Yodh AG, Contreras D, Salzberg BM (2008) Two-photon excitation of potentiometric probes enables optical recording of action potentials from mammalian nerve terminals in situ. J Neurophysiol 99:1545–1553

    Article  PubMed  Google Scholar 

  • Glover JC, Sato K, Momose-Sato Y (2008) Using voltage-sensitive dye recording to image the functional development of neuronal circuits in vertebrate embryos. Dev Neurobiol 68:804–816

    Article  PubMed  Google Scholar 

  • Grinvald A, Salzber BM, Cohen LB (1977) Simultaneous recording from several neurones in an invertebrate central nervous system. Nature 268:140–142

    Article  CAS  PubMed  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

    Article  CAS  PubMed  Google Scholar 

  • Grinvald A, Frostig RD, Lieke E, Hildesheim R (1988) Optical imaging of neuronal activity. Physiol Rev 68:1285–1366

    CAS  PubMed  Google Scholar 

  • Gupta RK, Salzberg BM, Grinvald A, Cohen LB, Kamino K, Lesher S, Boyle MB, Waggoner AS, Wang CH (1981) Improvements in optical methods for measuring rapid changes in membrane potential. J Membr Biol 58:123–137

    Article  CAS  PubMed  Google Scholar 

  • Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92

    Article  CAS  PubMed  Google Scholar 

  • Kamino K (1991) Optical approaches to ontogeny of electrical activity and related functional organization during early heart development. Physiol Rev 71:53–91

    CAS  PubMed  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

    Article  CAS  PubMed  Google Scholar 

  • Kamino K, Katoh Y, Komuro H, Sato K (1989) Multiple-site optical monitoring of neural activity evoked by vagus nerve stimulation in the embryonic chick brain stem. J Physiol (Lond) 409:263–283

    Article  PubMed Central  CAS  Google Scholar 

  • Loew LM (1988) How to choose a potentiometric membrane probe. In: Loew LM (ed) Spectroscopic membrane probes, vol 2. CRC Press, Boca Raton, pp 139–151

    Google Scholar 

  • Loew LM, Cohen LB, Dix J, Fluhler EN, Montana V, Salama G, Wu JY (1992) 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 Membr Biol 130:1–10

    Article  CAS  PubMed  Google Scholar 

  • Mochida H, Sato K, Arai Y, Sasaki S, Kamino K, Momose-Sato Y (2001) Optical imaging of spreading depolarization waves triggered by spinal nerve stimulation in the chick embryo: possible mechanisms for large-scale coactivation of the CNS. Eur J Neurosci 14:809–820

    Article  CAS  PubMed  Google Scholar 

  • Momose-Sato Y, Sato K (2011) The embryonic brain and development of vagal pathways. Respir Physiol Neurobiol 178:163–173

    Article  PubMed  Google Scholar 

  • Momose-Sato Y, Sato K (2013a) Optical imaging of the spontaneous depolarization wave in the mouse embryo: origins and pharmacological natures. Ann N Y Acad Sci 1279:60–70

    Article  CAS  PubMed  Google Scholar 

  • Momose-Sato Y, Sato K (2013b) Large-scale synchronized activity in the embryonic brainstem and spinal cord. Front Cell Neurosci 7:1–15

    Article  Google Scholar 

  • Momose-Sato Y, Sato K, Sakai T, Hirota A, Matsutani K, Kamino K (1995) Evaluation of optimal voltage-sensitive dyes for optical monitoring of embryonic neural activity. J Membr Biol 144:167–176

    Article  CAS  PubMed  Google Scholar 

  • Momose-Sato Y, Sato K, Arai Y, Yazawa I, Mochida H, Kamino K (1999) Evaluation of voltage-sensitive dyes for monitoring for long-term recording of neural activity in the hippocampus. J Membr Biol 172:145–157

    Article  CAS  PubMed  Google Scholar 

  • Momose-Sato Y, Sato K, Kamino K (2001) Optical approaches to embryonic development of neural functions in the brainstem. Prog Neurobiol 63:151–197

    Article  CAS  PubMed  Google Scholar 

  • Momose-Sato Y, Sato K, Kamino K (2002) Application of voltage-sensitive dyes to the embryonic central nervous system. In: Fagan J, Davidson JN, Shimizu N (eds) Recent research developments in membrane biology. Research Signpost, Kerala, pp 159–181

    Google Scholar 

  • Momose-Sato Y, Miyakawa N, Mochida H, Sasaki S, Sato K (2003) Optical analysis of large-scale depolarization waves in the embryonic brain: a dual network of gap junctions and chemical synapses. J Neurophysiol 89:600–614

    Article  PubMed  Google Scholar 

  • Obaid AL, Loew LM, Wuskell JP, Salzberg BM (2004) Novel naphthylstyryl-pyridinium potentiometric dyes offer advantages for neural network analysis. J Neurosci Meth 134:179–190

    Article  CAS  Google Scholar 

  • Onimaru H, Homma I (2003) A novel functional neuron group for respiratory rhythm generation in the ventral medulla. J Neurosci 23:1478–1486

    CAS  PubMed  Google Scholar 

  • Ross WN, Reichardt LF (1979) Species-specific effects on the optical signals of voltage-sensitive dyes. J Membr Biol 48:343–356

    Article  CAS  PubMed  Google Scholar 

  • Ross WN, Salzberg BM, Cohen LB, Grinvald A, Davila HV, Waggoner AS, Wang CH (1977) Changes in absorption, fluorescence, dichroism, and birefringence in stained giant axons: optical measurement of membrane potential. J Membr Biol 33:141–183

    Article  CAS  PubMed  Google Scholar 

  • Saggau P, Bullen A, Patel SS (1998) Acoustic-optic random-access laser scanning microscopy: fundamentals and applications to optical recording of neuronal activity. Cell Mol Biol 44:827–846

    CAS  PubMed  Google Scholar 

  • Salzberg BM (1983) Optical recording of electrical activity in neurons using molecular probes. In: Barker JL, McKelvy JF (eds) Current methods in cellular neurobiology, vol 3., Electrophysiological techniquesWiley, New York, pp 139–187

    Google Scholar 

  • Salzberg BM, Grinvald A, Cohen LB, Davila HV, Ross WN (1977) Optical recording of neuronal activity in an invertebrate central nervous system: simultaneous monitoring of several neurons. J Neurophysiol 40:1281–1291

    CAS  PubMed  Google Scholar 

  • Salzberg BM, Obaid AL, Senseman DM, Gainer H (1983) Optical recording of action potentials from vertebrate nerve terminals using potentiometric probes provides evidence for sodium and calcium components. Nature 306:36–40

    Article  CAS  PubMed  Google Scholar 

  • Sato K, Momose-Sato Y, Mochida H, Arai Y, Yazawa I, Kamino K (1999) Optical mapping reveals the functional organization of the trigeminal nuclei in the chick embryo. Neurosci 93:687–702

    Article  CAS  Google Scholar 

  • Sato K, Komuro R, Momose-Sato Y (2011) Screening of fluorescent voltage-sensitive dyes for detecting neural activity in the embryonic brain. J Physiol Sci 61(Suppl 1):S168

    Google Scholar 

  • Senseman DM, Salzberg BM (1980) Electrical activity in an exocrine gland: optical recording with a potentiometric dye. Science 208:1269–1271

    Article  CAS  PubMed  Google Scholar 

  • Wu JY, Cohen LB (1993) Fast multisite optical measurement of membrane potential. In: Mason WT (ed) Fluorescent and luminescent probes for biological activity. Academic, Boston, pp 389–404

    Google Scholar 

  • Yan P, Acker CD, Zhou W-L, Lee P, Bollensdorff C, Negrean A, Lotti J, Sacconi L, Antic SD, Kohl P, Mansvelder HD, Pavone FS, Loew LM (2012) Palette of fluorinated voltage-sensitive hemicyanine dyes. Proc Natl Acad Sci USA 109:20443–20448

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Zecevic D (1996) Multiple spike-initiation zones in single neurons revealed by voltage-sensitive dyes. Nature 381:322–325

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This research was supported by Grants from the Monbu-Kagaku-sho of Japan, the Human Frontier Science Program (Grant RGP0027/2009) and the US National Institutes of Health (grant R01EB001963). S. H. M. was supported by the Opto-Medical Institute as a postdoctoral fellowship.

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Authors and Affiliations

  1. Department of Health and Nutrition Sciences, Faculty of Human Health, Komazawa Women’s University, 238 Sakahama, Inagi-shi, Tokyo, 206-8511, Japan

    Saad Habib-E-Rasul Mullah & Katsushige Sato

  2. Department of Health and Nutrition, College of Human and Environmental Studies, Kanto Gakuin University, Yokohama, 236-8501, Japan

    Ryo Komuro & Yoko Momose-Sato

  3. R. D. Berlin Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT, 06030-6406, USA

    Ping Yan & Leslie M. Loew

  4. Department of Neurosurgery, Graduate School and Faculty of Medicine, Tokyo Medical and Dental University, Tokyo, 113-8519, Japan

    Shihori Hayashi & Motoki Inaji

Authors
  1. Saad Habib-E-Rasul Mullah
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  2. Ryo Komuro
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  3. Ping Yan
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  4. Shihori Hayashi
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  5. Motoki Inaji
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  6. Yoko Momose-Sato
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  7. Leslie M. Loew
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  8. Katsushige Sato
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Correspondence to Katsushige Sato.

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Habib-E-Rasul Mullah, S., Komuro, R., Yan, P. et al. Evaluation of Voltage-Sensitive Fluorescence Dyes for Monitoring Neuronal Activity in the Embryonic Central Nervous System. J Membrane Biol 246, 679–688 (2013). https://doi.org/10.1007/s00232-013-9584-1

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  • DOI: https://doi.org/10.1007/s00232-013-9584-1

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