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Techniques for Studying Neuromast Function in Zebrafish

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Flow Sensing in Air and Water

Abstract

The mechano-sensitive hair cells of superficial neuromasts (SNs) of the zebrafish lateral line organ are mechanically coupled to the water motion via gelatinous cupulae. SNs transduce the water motion into electrical signals that can be measured with an extracellular electrode. In this chapter, we review the preparation and measurement techniques for quantifying cupular dynamics and extracellular receptor potentials (ERPs) of SNs. We compare the measuring techniques used in hair cell mechano-physiology and give instructions for building both an intensity-based and an interferometry-based microscope system. We compare the methods used for mechanical excitation of mechanoreceptors, including dipole sources, microfluid jets (FJ) and elastic as well as stiff microprobes. We present the caveats of the measurements of ERPs, especially the crosstalk from the stimulation device. We show that ERPs at twice the stimulation frequency of zebrafish SNs are a reliable measure of mechano-electrical coupling in a restricted range of both stimulus frequency and amplitude. We report the measurements of sub-micrometre motion of SN cupulae using a heterodyne laser interferometer microscope (HLIM) and continuous sinusoidal stimulation with a micro FJ device. Light interference signals were decoded with a phase- and frequency modulation scheme. We compare the robustness of both decoding strategies in terms of accuracy of the measured cupular displacement and velocity. Both approaches can faithfully monitor cupular movement down to a few nanometres, though the velocity decoding technique offered a slightly superior performance and is recommended for higher stimulation frequencies.

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Abbreviations

CCD:

Charge-coupled device

CMOS:

Complementary metal oxide semiconductor

CN:

Canal neuromast

DIC:

Differential interference contrast

ERP:

Extracellular receptor potential

FFT:

Fast Fourier transform

FJ:

Fluid jet (device)

HLIM:

Heterodyning laser interferometer microscopy/microscope

LED:

Light emitting diode

MIPO:

Microphonic potential

PD:

Photodiode

PMT:

Photomultiplier tube

PSD:

Position sensitive device

RMS:

Root mean square

SN:

Superficial neuromast

SNR:

Signal-to-noise ratio

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Acknowledgments

The authors wish to thank the reviewers and the editors for their insightful comments. The work presented in this chapter was chiefly funded by the EU FP6 Project Cilia.

Author information

Authors and Affiliations

  1. Department of Materials and Metallurgy, Faculty of Natural Sciences and Engineering, University of Ljubljana, Aškerčeva 12, SI-1000, Ljubljana, Slovenia

    Primož Pirih

  2. Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia

    Primož Pirih

  3. Inserm U1051-Institute for Neurosciences of Montpellier, Montpellier, France

    Gaston C. Sendin

  4. Institute of Artificial Intelligence and Cognitive Engineering, University of Groningen, Groningen, The Netherlands

    Sietse M. van Netten

Authors
  1. Primož Pirih
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  2. Gaston C. Sendin
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  3. Sietse M. van Netten
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Corresponding author

Correspondence to Primož Pirih .

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Editors and Affiliations

  1. Institute for Zoology, University of Bonn, Bonn, Germany

    Horst Bleckmann

  2. Institute for Zoology, University of Bonn, Bonn, Germany

    Joachim Mogdans

  3. Department of Biological Sciences, Bowling Green State University, Bowling Green, Ohio, USA

    Sheryl L. Coombs

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Pirih, P., Sendin, G.C., van Netten, S.M. (2014). Techniques for Studying Neuromast Function in Zebrafish. In: Bleckmann, H., Mogdans, J., Coombs, S. (eds) Flow Sensing in Air and Water. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-41446-6_14

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