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Mitochondrial Ca2+ mobilization is a key element in olfactory signaling

Nature Neuroscience volume 15, pages 754–762 (2012)Cite this article

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Abstract

In olfactory sensory neurons (OSNs), cytosolic Ca2+ controls the gain and sensitivity of olfactory signaling. Important components of the molecular machinery that orchestrates OSN Ca2+ dynamics have been described, but key details are still missing. Here, we demonstrate a critical physiological role of mitochondrial Ca2+ mobilization in mouse OSNs. Combining a new mitochondrial Ca2+ imaging approach with patch-clamp recordings, organelle mobility assays and ultrastructural analyses, our study identifies mitochondria as key determinants of olfactory signaling. We show that mitochondrial Ca2+ mobilization during sensory stimulation shapes the cytosolic Ca2+ response profile in OSNs, ensures a broad dynamic response range and maintains sensitivity of the spike generation machinery. When mitochondrial function is impaired, olfactory neurons function as simple stimulus detectors rather than as intensity encoders. Moreover, we describe activity-dependent recruitment of mitochondria to olfactory knobs, a mechanism that provides a context-dependent tool for OSNs to maintain cellular homeostasis and signaling integrity.

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Figure 1: Bioluminescence imaging of mitochondrial Ca2+ dynamics in the stimulated mouse MOE.
Figure 2: Mitochondria regulate Ca2+ shuttling during odor responses.
Figure 3: [Ca2+]m mobilization shapes [Ca2+]c signals.
Figure 4: [Ca2+]m dynamics exert local effects.
Figure 5: Mitochondrial Ca2+ uptake ensures a broad dynamic range.
Figure 6: Action potential output in Olfr73-expressing OSNs is regulated by mitochondrial Ca2+ buffering.
Figure 7: Activity-dependent mitochondrial transport.
Figure 8: Recruitment of mitochondria to OSN knobs.

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Acknowledgements

We thank C. Engelhardt, H. Bartel and S. Lipartowski for assistance, W. Kammerloher and H.-J. Behrendt (Olympus Life Science) for placing the LV200 microscope at our disposal, and K. Touhara (University of Tokyo, Japan) and P. Mombaerts (Max Planck Institute of Biophysics, Frankfurt, Germany) for providing mouse strains. This work was funded by grants from the Volkswagen Foundation (M.S.) and the Deutsche Forschungsgemeinschaft (M.S., SP724/2-1; E.M.N., Exc257). M.S. is a Lichtenberg Professor of the Volkswagen Foundation.

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  1. Daniela Fluegge and Lisa M Moeller: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemosensation, Institute for Biology II, RWTH (Rheinisch-Westfälische Technische Hochschule) Aachen University, Aachen, Germany

    Daniela Fluegge, Lisa M Moeller, Annika Cichy, Monika Gorin, Sophie Veitinger, Jennifer Spehr & Marc Spehr

  2. Department of Cellular Neurobionics, Institute for Biology II, RWTH Aachen University, Aachen, Germany

    Agnes Weth & Werner Baumgartner

  3. Axxam SpA, Milan, Italy

    Silvia Cainarca, Stefan Lohmer & Sabrina Corazza

  4. Charité–NeuroScience Research Center, Cluster of Excellence NeuroCure, Berlin, Germany

    Eva M Neuhaus

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Contributions

Experiments were performed in the laboratories of M.S., W.B. and S.L. Original concept of research: D.F., S.V., S.L., E.M.N., J.S. and M.S. Research was designed by D.F., S.V., S. Cainarca, S. Corazza, E.M.N., W.B., J.S. and M.S. Data were collected by D.F., L.M.M., A.C., M.G., A.W., S.V. and J.S. and analyzed by D.F., L.M.M., J.S. and M.S. The manuscript was written by D.F., J.S. and M.S. (assistance from L.M.M., S.V., S. Corazza, W.B., E.M.N.).

Corresponding author

Correspondence to Marc Spehr.

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Competing interests

The project was supported by Axxam SpA. S. Cainarca, S.L. and S. Corazza were full-time employees of Axxam SpA during project conception and data generation and were involved in study design, data collection and analysis, and decision to publish. S.L. also has a personal financial interest in Axxam SpA as cofounder and shareholder. The following three patent applications (owned by Axxam SpA) are relevant to work described in the paper:

1. European Patent 06000452.0, World Intellectual Property Organization Patent 2007080622 (19 July 2007).

2. European Patent 06000452.0, World Intellectual Property Organization Patent 2007080621 (19 July 2007).

3. World Intellectual Property Organization Patent 2006094805 (14 September 2006); priority numbers: European Patent 20050005390 20050311, European Patent 20060000171 20060105.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10 (PDF 12769 kb)

Supplementary Video 1

Bioluminescence imaging of mitochondrial Ca2+ dynamics in the stimulated mouse MOE. Acute coronal slices (150 μm thick) of the rostral skull of PhotoTopo mice at postnatal days P2 (Video 1), P4 (Video 2), and P6 (Video 3) are visualized at 20x magnification. For spatial orientation, 'static' transmitted light images are merged with the original frames (1 Hz capture rate) of representative live-cell [Ca2+]m bioluminescence time-lapse recordings. Bioluminescence intensity is presented in pseudocolor (transparent grey = low [Ca2+]m, red = high [Ca2+]m). Slice superfusion with a complex odor mixture (100 compounds, ~10 μM each; 10 s) induces transient light emission in spatially confined regions of the sensory epithelium, whereas bioluminescence responses are not observed in the respiratory epithelium. Membrane depolarization by an elevated extracellular K+ concentration (50 mM) triggers bioluminescent signals in broader MOE areas. By simultaneously monitoring large epithelial areas at low magnification, a gradual response delay along a ventromedial-dorsolateral axis becomes apparent as a function of epithelial distance from the perfusion pencil. (MP4 4957 kb)

Supplementary Video 2

See Supplementary Video 1 (MP4 6863 kb)

Supplementary Video 3

See Supplementary Video 1 (MP4 5959 kb)

Supplementary Video 4

Confocal time-lapse fluorescence imaging of [Ca2+]c responses in the mouse MOE. Coronal MOE slices are bulk loaded with a Ca2+-sensitive dye (fluo-4/AM; 2 μM) and monitored at high magnification. Here, single OSNs are readily discernible. Fluorescence frame sequences are merged with a 'static' confocal DIC image of the MOE area under investigation to show an anatomical reference. To provide a high contrast display of fluorescence changes, averaged 'baseline' images were subtracted from each original frame, thus generating a time-lapse movie in which only deviations from baseline fluorescence (ΔF) become discernible as pseudocolor intensity changes (red = elevated [Ca2+]c). Note, however, that unavoidable miniature movements of the slice (in the nanometer range) during the course of an experiment result in halo artifacts. These 'halos' stem from the subtraction of a 'baseline' image that is not perfectly congruent with a respective frame. Movies show (i) both odor- and K+-dependent [Ca2+]c signals (Video 4), or (ii) K+-triggered [Ca2+]c transients (Video 5) in single OSNs under control conditions. Brief odor stimulation triggers [Ca2+]c transients in a subpopulation of OSNs, whereas K+-mediated depolarization activates the majority of neurons. (MP4 684 kb)

Supplementary Video 5

See Supplementary Video 4 (MP4 747 kb)

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Fluegge, D., Moeller, L., Cichy, A. et al. Mitochondrial Ca2+ mobilization is a key element in olfactory signaling. Nat Neurosci 15, 754–762 (2012). https://doi.org/10.1038/nn.3074

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