ATP Consumption of Eukaryotic Flagella Measured at a Single-Cell Level

Daniel T.N. Chen, Michael Heymann, Seth Fraden, Daniela Nicastro, Zvonimir Dogic

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The motility of cilia and flagella is driven by thousands of dynein motors that hydrolyze adenosine triphosphate (ATP). Despite decades of genetic, biochemical, structural, and biophysical studies, some aspects of ciliary motility remain elusive, such as the regulation of beating patterns and the energetic efficiency of these nanomachines. In this study, we introduce an experimental method to measure ATP consumption of actively beating axonemes on a single-cell level. We encapsulated individual sea urchin sperm with demembranated flagellum inside water-in-oil emulsion droplets and measured the axoneme's ATP consumption by monitoring fluorescence intensity of a fluorophore-coupled reporter system for ATP turnover in the droplet. Concomitant phase contrast imaging allowed us to extract a linear dependence between the ATP consumption rate and the flagellar beating frequency, with ∼2.3 × 105 ATP molecules consumed per beat of a demembranated flagellum. Increasing the viscosity of the aqueous medium led to modified beating waveforms of the axonemes and to higher energy consumption per beat cycle. Our single-cell experimental platform provides both new insights, to our knowledge, into the beating mechanism of flagella and a powerful tool for future studies.

Original languageEnglish (US)
Pages (from-to)2562-2573
Number of pages12
JournalBiophysical Journal
Volume109
Issue number12
DOIs
StatePublished - Dec 15 2015

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Flagella
Adenosine Triphosphate
Axoneme
Dyneins
Sea Urchins
Cilia
Emulsions
Viscosity
Spermatozoa
Molecular Biology
Oils
Fluorescence
Water

ASJC Scopus subject areas

  • Biophysics

Cite this

ATP Consumption of Eukaryotic Flagella Measured at a Single-Cell Level. / Chen, Daniel T.N.; Heymann, Michael; Fraden, Seth; Nicastro, Daniela; Dogic, Zvonimir.

In: Biophysical Journal, Vol. 109, No. 12, 15.12.2015, p. 2562-2573.

Research output: Contribution to journalArticle

Chen, Daniel T.N. ; Heymann, Michael ; Fraden, Seth ; Nicastro, Daniela ; Dogic, Zvonimir. / ATP Consumption of Eukaryotic Flagella Measured at a Single-Cell Level. In: Biophysical Journal. 2015 ; Vol. 109, No. 12. pp. 2562-2573.
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