Data/Software for "Presynaptic Mitochondria Volume and Abundance Increase During Development of a High-Fidelity Synapse"
Connon I. Thomas,
Connon I. ThomasMax Planck Florida Institute for Neuroscience
- Christian Keine,
Satoko OkayamaMax Planck Florida Institute for Neuroscience
Rachel SatterfieldMax Planck Florida Institute for Neuroscience
Morgan MusgroveMax Planck Florida Institute for Neuroscience
Debbie Guerrero-GivenMax Planck Florida Institute for Neuroscience
Naomi KamasawaMax Planck Florida Institute for Neuroscience
Samuel M. Young, Jr.
Samuel M. Young, Jr.University of Iowa
Description of this data
Contains data and software from the publication: "Presynaptic Mitochondria Volume and Abundance Increase During Development of a High-Fidelity Synapse" published in the The Journal of Neuroscience (https://doi.org/10.1523/JNEUROSCI.0363-19.2019).
The preprint to this data set has been published on bioRxiv (https://doi.org/10.1101/689653).
In this study, we created a helper-dependent adenoviral vector (HdAd) to co-express cytoplasmic EGFP and a genetically encoded peroxidase marker (mito-APEX2) at the calyx of Held, an excellent model for deciphering regulatory mechanisms of presynaptic function.
The calyx of Held, a large glutamatergic presynaptic terminal in the auditory brainstem undergoes developmental changes to support the high action-potential firing rates required for auditory information encoding. In addition, calyx terminals are morphologically diverse which impacts vesicle release properties and synaptic plasticity. Mitochondria influence synaptic plasticity through calcium buffering and are crucial for providing the energy required for synaptic transmission. Therefore, it has been postulated that mitochondrial levels increase during development and contribute to the morphological-functional diversity in the mature calyx. However, the developmental profile of mitochondrial volumes and subsynaptic distribution at the calyx of Held remains unclear. To provide insight on this, we developed a helper-dependent adenoviral vector (HdAd) that expresses the genetically encoded peroxidase marker for mitochondria, mito-APEX2, at the mouse calyx of Held. We developed protocols to detect labeled mitochondria for use with serial block face scanning electron microscopy to carry out semi-automated segmentation of mitochondria, high-throughput whole terminal reconstruction and presynaptic ultrastructure in mice of either sex. Subsequently, we measured mitochondrial volumes and subsynaptic distributions at the immature postnatal day 7 (P7) and the mature (P21) calyx. We found an increase of mitochondria volumes in terminals and axons from P7 to P21 but did not observe differences between stalk and swelling subcompartments in the mature calyx. Based on these findings, we propose that mitochondrial volumes and synaptic localization developmentally increase to support high firing rates required in the initial stages of auditory information processing.
Data are sorted by the figures they appear in. Media (movies and 3D models) and custom-written software are located in separate folders.
Experiment data files
custom-written software used for analysis for Matlab (RRID:SCR_001622) and Python (RRID:SCR_008394)
Functions used in Matlab 2018a and newer, but should work in older versions as well (>2013b).
Class to import HEKA Patchmaster files into Matlab. Please see the file description and Readme_HEKA_Importer.txt for further details. This is the version used at the time point of publication, please see https://github.com/ChristianKeine/HEKA_Patchmaster_Importer for updates.
Functions used in Python 3, not tested for use in Python 2
Data sets for Figures
Data sorted by Figure in which they are displayed. Data are stored as CSV format. Original images are stored as TIFF.
Data for Figure 1: Expression of EGFP-mito-APEX2 at the calyx of Held terminal.
Data for Figure 2: Double labeling of EGFP and mito-APEX2 using a combination of pre-embedding DAB-reaction labeling and post-embedding immunogold labeling.
Data for Figure 3: Expression of mito-APEX2 does not affect synaptic transmission at the calyx of Held-MNTB synapse.
Raw data files recorded with HEKA Patchmaster. Can be loaded into Patchmaster or imported to Matlab using functions from Data > Code/software > Matlab > @HEKA_Importer.
Data for Figure 4: Mito-APEX2 labeling visualized by SBF-SEM at P7 (A, B) and P21 (C, D, E1) allows for identification of transduced calyx of Held terminals
Data for Figure 5: Large volume reconstructions of mito-APEX2-positive calyx of Held terminals using SBF-SEM at P7 and at P21.
Data for Figure 6: Volumetric reconstructions of mitochondria from the same whole terminals and axons shown in Figure 5 using SBF-SEM at P7 and P21.
Data for Figure 7: Full resolution reconstructions of sub-compartments and mitochondria of calyx of Held terminals at P7 and P21 using SBF-SEM.
Data for Figure 8: Mitochondria and membrane surface details of a segment of mito-APEX2-positive calyx of Held terminal reconstructed at full resolution from a SBF-SEM dataset at P7.
Data for Figure 9: Mito-APEX2 labeling visualized by ssSEM at P7 (A, B) and P21 (C, D, E1) allows for identification of transduced calyx of Held terminals.
Data for Figure 10: Mito-APEX2 labeling visualized by ssSEM at P7 and P21 allows for identification of transduced calyx of Held terminals as well as subsequent 3D synaptic analysis.
Movies and 3D models
Movies and 3D models of reconstructions.
3D reconstructions. Stored as *.obj, *.blender (RRID:SCR_008606), or Amira (RRID:SCR_014305) files.
Movies of 3D reconstructions stored as MP4 format.
Steps to reproduce
All data are stored in CSV-files with periods as decimal point and columns separated by commas. The first line in the CSV file corresponds to the header which identifies the groups.
This data is associated with the following publication:
Cite this dataset
Thomas, Connon I.; Keine, Christian; Okayama, Satoko; Satterfield, Rachel; Musgrove, Morgan; Guerrero-Given, Debbie; Kamasawa, Naomi; Young, Jr., Samuel M. (2019), “Data/Software for "Presynaptic Mitochondria Volume and Abundance Increase During Development of a High-Fidelity Synapse"”, Mendeley Data, v4 http://dx.doi.org/10.17632/v88r5t5myz.4
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The files associated with this dataset are licensed under a Creative Commons Attribution 4.0 International licence.