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Release probability of hippocampal glutamatergic terminals scales with the size of the active zone

Nature Neuroscience volume 15, pages 988–997 (2012)Cite this article

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A Corrigendum to this article was published on 29 December 2015

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Abstract

Cortical synapses have structural, molecular and functional heterogeneity; our knowledge regarding the relationship between their ultrastructural and functional parameters is still fragmented. Here we asked how the neurotransmitter release probability and presynaptic [Ca2+] transients relate to the ultrastructure of rat hippocampal glutamatergic axon terminals. Two-photon Ca2+ imaging–derived optical quantal analysis and correlated electron microscopic reconstructions revealed a tight correlation between the release probability and the active-zone area. Peak amplitude of [Ca2+] transients in single boutons also positively correlated with the active-zone area. Freeze-fracture immunogold labeling revealed that the voltage-gated calcium channel subunit Cav2.1 and the presynaptic protein Rim1/2 are confined to the active zone and their numbers scale linearly with the active-zone area. Gold particles labeling Cav2.1 were nonrandomly distributed in the active zones. Our results demonstrate that the numbers of several active-zone proteins, including presynaptic calcium channels, as well as the number of docked vesicles and the release probability, scale linearly with the active-zone area.

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Figure 1: Morphological diversity of glutamatergic axon terminals in the CA3 region of the rat hippocampus.
Figure 2: The number of docked vesicles correlates linearly with active-zone size.
Figure 3: Determining the release probability of axon terminals with optical quantal analysis.
Figure 4: Post hoc ultrastructural analysis of synapses after two-photon imaging.
Figure 5: Measurement of volume-averaged [Ca2+] transients in CA3 pyramidal cell local axon terminals.
Figure 6: Cav2.1 subunit was confined to the active zone of presynaptic axon terminals in the stratum oriens of the hippocampal CA3 area.
Figure 7: The number of Cav2.1 subunits and Rim1/2 proteins correlates with the active-zone area.
Figure 8: Nonrandom distribution of the Cav2.1 subunits within the active zones.

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  • 23 November 2015

    In the version of this article initially published, Figure 7h presented data from rat 2 but the corresponding legend gave statistics for rat 1. The legend originally read, “Density of gold particles labeling the Cav2.1 subunit within presynaptic active zones (mean ± s.d. = 395.8 ± 154.8 gold μm−2, n = 34 in rat 1) and in the surrounding extrasynaptic axonal plasma membrane (mean ± s.d. = 1.6 ± 2.4 gold μm−2, n = 32 in rat 1) in comparison with the background labeling calculated on E-face plasma membranes (mean ± s.d. = 0.6 ± 2.3 gold μm−2, n = 39; Psynaptic < 0.01, Pextrasynaptic = 0.73).” It has been changed to give the statistics for rat 2: “Density of gold particles labeling the Cav2.1 subunit within presynaptic active zones (mean ± s.d. = 293.8 ± 122 gold μm−2, n = 49 in rat 2) and in the surrounding extrasynaptic axonal plasma membrane (mean ± s.d. = 2.8 ± 4.0 gold μm−2, n = 49 in rat 2) in comparison with the background labeling calculated on E-face plasma membranes (mean ± s.d. = 0.33 ± 1.2 gold μm−2, n = 57; Psynaptic < 0.01, Pextrasynaptic = 0.06).” The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

N.H. and A.L. are funded by Janos Bolyai Scholarships of the Hungarian Academy of Sciences. Z.N. is supported by Wellcome Trust Equipment (083484/Z/07/Z) and Project Grants (090197/Z/09/Z; 094513/Z/10/Z), a European Research Council Advanced Grant, and a Hungarian National Office for Research and Technology-French National Research Agency TéT Fund (NKTH-Neurogen). B.R. was supported by a GOP grant (1.1.1-08/1-2008-0085). A.K. is supported by a Deutsche Forschungsgemeinschaft (SFB 780) grant. We thank N. Suzuki (Mie University, Japan) for providing Cav2.1−/− mice, A. Unger for helping with the Cav2.1−/− replica labeling, E. Dobai and D. Ronaszéki for technical assistance, M. Sümegi for his help with the modeling, members of Synaptic Systems GmbH for providing the rabbit anti-Cav2.1 antibody, E. Neher and M. Eyre for their comments on the manuscript, and H. Koester for helpful discussions.

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Authors and Affiliations

  1. Laboratory of Cellular Neurophysiology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary

    Noemi Holderith, Andrea Lorincz & Zoltan Nusser

  2. Two-Photon Imaging Center, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary

    Gergely Katona & Balázs Rózsa

  3. Department of Neuroanatomy, Department of Physiology II, Institute of Anatomy and Cell Biology, University of Freiburg, Freiburg, Germany

    Akos Kulik

  4. Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany

    Akos Kulik

  5. Department of Anatomy, Hokkaido University School of Medicine, Sapporo, Japan

    Masahiko Watanabe

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  1. Noemi Holderith
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Contributions

N.H. performed all in vitro Ca2+ measurements and post hoc light- and electron microscopy analysis of the imaged structures. A.L. performed SDS-FRL for SNAP-25, Cav2.1 and Rim1/2, quantitatively analyzed reactions and performed simulations. G.K. developed the software. B.R. designed and built the two-photon microscope. A.K. performed SDS-FRL reactions for Cav2.1 in Cav2.1+/− and Cav2.1−/− mice. M.W. developed the guinea pig anti-Cav2.1 antibody. N.H. and Z.N. designed experiments and wrote the manuscript.

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Correspondence to Zoltan Nusser.

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B.R. and G.K. are the owners of Femtonics Ltd.

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Holderith, N., Lorincz, A., Katona, G. et al. Release probability of hippocampal glutamatergic terminals scales with the size of the active zone. Nat Neurosci 15, 988–997 (2012). https://doi.org/10.1038/nn.3137

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