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26 results for “exocytosis”
Receptor exocytosis imaged with high temporal resolution for diverse receptor cargos
<p>Cells perceive and interact with their environment in part through the expression, activation, and regulation of receptors on their plasma membrane. These receptors are dynamically trafficked from the plasma membrane in a process called endocytosis and delivered to the plasma membrane via exocytosis. Different receptors take diverse routes through the cell before being delivered via exocytosis. The data in this project focuses on 3 prototypical plasma membrane receptors - the B2 adrenergic receptor, the µ opioid receptor, and the transferrin receptor. Using a pH-sensitive green fluorescent protein variant, we visualized these receptors in cells as they recycled to the plasma membrane. We subsequently hand-labeled a subset of the data in order to build an automated image analysis method that could be used to detect receptor exocytosis across diverse imaging conditions. This repository contains our primary microscopy data from these studies as well as the labeling for use in supervised machine learning.</p> <p>These data support <a href="http://arxiv.org/abs/2106.07623">Evans et al 2021</a> and subsequent publications.</p> <p><strong>Data Collection</strong><br> TIFF image stacks were collected using a Nikon Eclipse TiE Inverted Microscope using TIRF illumination with a solid state 488nm laser through a Nikon 60x/1.49NA TIRF objective and captured using an Andor iXon 897+ EMCCD camera. The camera was windowed to a 300x300 pixel view and images were collected with a 18.5ms exposures (~54Hz). Images were collected across two days, with two coverslips of each condition collected on day 1, and one coverslip collected on day 2.</p> <p><strong>DNA Constructs</strong><br> The 3 cargos imaged in these data are the transferrin receptor (TfR), the B2-adrenergic receptor (B2AR, B2), and the µ opioid receptor (MOR). Constructs encoding these receptors, tagged extracellularly with the ph-sensitive GFP variant Superecliptic pHluorin (SpH, <a href="https://www.cell.com/biophysj/fulltext/S0006-3495(00)76468-X">Sankaranarayanan et al. 2000</a>, have been previously described in <a href="http://www.nature.com/articles/nn1679">Yudowski et al. 2006</a> for B2AR, <a href="https://www.jneurosci.org/content/30/35/11703">Yu et al. 2010</a> for MOR, and <a href="https://www.molbiolcell.org/doi/10.1091/mbc.e08-08-0892">Yudowski et al. 2009</a> for TfR.</p> <p><strong>Cell Culture</strong><br> HEK293 cells were cultured in DMEM High Glucose (Hyclone) supplemented with 10% Heat Inactivated FBS (Gibco). Cells expressing B2 and MOR were stably selected from transient transfection using G418. Cells expressing TfR were transfected 3 days before the experiments presented here using Effectene following manufacturers' instructions. Before imaging, cells were transferred to 25mm diameter #1.5 glass coverslips (Electron Microscopy Sciences). Two days after plating, experiments began.</p> <p><strong>Imaging conditions</strong><br> Cells were imaged in L-15 minimal media supplemented with 1% FBS. For MOR and B2, cells were imaged for 1 minute at ~0.16Hz without perturbation. Then agonist was added (10µM DAMGO for MOR, 10µM isoproterenol for B2) to the media and cells were imaged for 5 minutes to ensure that receptors clustered and internalized. After internalization, cells were bleached with 100% laser power for 1 minute and then imaged at 54Hz to visualize exocytic events. exocytosis was captured for up to 20 minutes after initial treatment, one cell at a time. For TfR, a single frame was taken before bleaching to show receptor expression levels and then cells were bleached and imaged as described above.</p> <p><strong>Data blinding</strong><br> After collection, files were renamed as described in <em>map.md</em>. All metadata files and internalization imaging were separated into the 2 "extras" folders. The exocytosis movies were 'scrambled' to hide cargo identity using the included <em>scrambler.py</em> file. <em>OPP_scramble.log</em> described the mapping of scrambled filenames to the original imaging.</p> <p><strong>Human labeling</strong><br> A subset of the images (22, with roughly equal representation across cargos) were hand labeled for exocytic events. Images were viewed in FIJI <a href="https://www.nature.com/articles/nmeth.2019">Schindelin et al. 2012</a> nad played back at 0.5x. When exocytic events were identified by eye, the playback was paused and the appearance of an event was found through manual advancing of the frames of the movie. The event was labeled using the Cell Counter plugin. Each movie was watched twice to identify as many events as possible. Labeled events are saved a <em><movie-name>-ZYW-1.xml</em> in this dataset.</p> <p><strong>Data organization</strong><br> All exocytic event movies and any matching human labeling are included in this base directory. All internalization movies and all metadata for all movies are included in the Extras folder for the day that movie was recorded. Coverslip and cargo identity are listed in <em>map.md</em> and the ground truth for cargo identity is in <em>OPP_scramble.log</em></p>
Unique dynamics and exocytosis properties of GABAergic synaptic vesicles revealed by three-dimensional single vesicle tracking
<p>This data set includes x, y, and z trajectories of all GABAergic synaptic vesicles that we used for the study. These GABAergic synaptic vesicles in inhibitory presynaptic terminals of living primary hippocampal neurons were labeled by single quantum dots (QDs) conjugated with anti-VGAT antibody under electrical stimulation, and were tracked three-dimensionally by using a dual-focus imaging in real-time. Each trajectory data indicates x, y, and z positions (nanometer-scale) over time from the start of imaging to the moment of vesicle fusion. The electrical stimulation to the neurons was applied for 120 s, starting from 20 s.</p>
Data for: Exocytosis of the silicified cell wall of diatoms involves extensive membrane disintegration
<p>Diatoms are unicellular algae, characterized by silica cell walls. The silica elements are formed intracellularly in a membrane-bound silica deposition vesicle (SDV), and are exocytosed after completion. How diatoms maintain membrane homeostasis during the exocytosis of these large and rigid silica elements is a long-standing enigma. We studied membrane dynamics during cell wall formation and exocytosis in two model diatom species, using live-cell confocal microscopy, transmission electron microscopy and cryo-electron tomography. Our results show that during the formation of the mineral phase it is in tight association with the SDV membranes, which are forming a precise mold of the delicate geometrical patterns. During exocytosis, the distal SDV membrane and the plasma membrane gradually detach from the mineral and disintegrate in the extracellular space, without any noticeable endocytic retrieval or extracellular repurposing. Within the cell, there is no evidence for the formation of a new plasma membrane, thus the proximal SDV membrane becomes the new barrier between the cell and its environment, and assumes the role of a new plasma membrane. These results provide direct structural observations of diatom silica exocytosis, and point to an extraordinary mechanism in which membrane homeostasis is maintained by discarding, rather than recycling, significant membrane patches.</p>
Figs 1–7 in Exocytosis of fibrous material from plasmatocytes in Scutigera coleoptrata (Chilopoda, Notostigmophora) in relation to wound healing
Figs 1–7. Plasmatocytes of Scutigera coleoptrata.1. Plasmatocyte (pl) with several grana (g) containing fibrous material. The hemocyte is attached to an axon (a). 2. Numerous grana, containing tubular fibrous substances and electron dense areas (ea). 3. Details of a granulum with an electron dense area. The arrows show a marginal detachment of tubuli from that centre; grana membrane (m). 4. Grana and areas where grana membranes (m) are dissolved; note that the fibrous tubuli remain ordered. 5. Cross sectioned fibrous tubuli (asterisk), (m) membrane. 6. Plasmatocyte, releasing tubuli in clusters (tc) into the hemolymph (h), nucleus (n). 7. Plasmatocyte, releasing tubuli singly into hemolymph. Tubuli in longitudinal section (long arrow), tubuli in cross section (short arrow). Asterisk indicates cross-sectioned tubuli.
Data for: Exocytosis of the silicified cell wall of diatoms involves extensive membrane disintegration
Open the record for dataset details and reuse information.
Mouse sperm acrosome exocytosis
<p>CD1 male mice (10 to 12 weeks old) were maintained in a 12-hour light and 12-hour dark cycle, at 23°C and 55±15% humidity, with water being always accessible. Animals were euthanized and cauda epididymal sperm collection followed. Cauda epididymis was cut at multiple sites, placed in 500 µl of non-capacitating medium (NC) and incubated at 37°C for 15 min. Supernatant was collected and pre-incubation with 100 nM SiR-actin in NC of recovered sperm took place for 10 min. Once dyed, sperm were once more incubated for at 37°C for 60 min in capacitating (CAP) conditions.</p> <p>NC used was a modified TYH medium (119.3 mM NaCl, 4.7 mM KCl, 1.71 mM CaCl<sub>2</sub>•2H<sub>2</sub>O, 1.2 mM KH<sub>2</sub>PO<sub>4</sub>, 1.2 mM MgSO<sub>4•</sub>7H<sub>2</sub>O, 0.51 mM sodium pyruvate, 5.56 mM glucose, 20 mM HEPES and 10 µg/ml gentamicin). For CAP conditions, 5 mg/ml BSA and 15 mM NaHCO<sub>3</sub> were added.</p> <p>Sperm were immobilized in coverslips treated with concanavalin-A (1 mg/ml). The imaging chamber was loaded with NC with 0.5 µM FM4-64 and 100 nM SiR-actin. Dye excitation was provided by 561 nm and 640 nm lasers. Using a NanoImager-S microscope (ONI, Oxford Nanoimaging Ltd) equipped with a 100X, 1.4 NA, oil-immersion objective (Olympus), 100 frames were acquired every 0.5 min for a total of 20 min, with a pixel size of 117 nm.</p> <p>Experimental procedures were approved by the Bioethics Committee of the Biotechnology Institute of the National Autonomous University of Mexico.</p>
TIRF imaging of lysosomal exocytosis
<p><strong><span> </span></strong><strong><span>1\ Training Dataset (120 movies)</span></strong></p> <p><span>This dataset contains the data used for the training and the evaluation of ExoDeepFinder (i.e. the 120 movies presented in figure 1D). Each folder corresponds to a single TIRFM movie of one RPE1 VAMP7-pHluorin transfected cell. All cells are in control conditions <em>i.e. </em>no drug treatment. Importantly, some movies are from paired experiments, before-after treatment, but only conditions before treatment have been included in the dataset. Hence, the mentioning of a drug in the folder name is a reference to the control before the aforementioned treatment and does not indicate drug treatment. While most of the cells are seeded on fibronectin-coated coverslips (62 movies), some of them are seeded on different substrates:</span></p> <p><span>-Cells from folders with the reference “Micropattern Ring” were seeded on fibronectin coated, ring-shaped micropatterns (diameter of 37µm and thickness of the adhesive ring of 7µm) (17 movies).</span></p> <p><span>-Cells from folders with the reference “Rectangles pattern” or “Micropattern rectangle” were seeded on fibronectin coated, rectangle-shaped micropatterns (9x40µm) (18 movies).</span></p> <p><span>-Cells from folders with the reference “PLL coating” were seeded on PLL coated coverslips (23 movies).</span></p> <p><span>Moreover, while most of the cells are only VAMP7-pHluorin transfected (96 movies), some of them have been co-transfected:</span></p> <p><span>-Cells from folders with the reference “Paxillin” were co-transfected with paxillin-mCh (19 movies).</span></p> <p><span>-Cells from folders with the reference “RFP-Rab6A” were co-transfected with mCh-Rab6A (5 movies).</span></p> <p><span>Each folder contains a sequence of .TIF images corresponding to each frame of the movie. Movies can be opened with ImageJ as an image sequence. Each movie is made of 1001 16-bit images. The pixel size is 0.160µm. In addition, each folder contains a .nd file that can be opened with ImageJ. Moreover, a .tif cell mask is associated to each movie (“Flood fill” to fill holes inside the mask and “Clear outside” algorithms are not applied on these masks but necessary to obtain simply connected masks). </span></p> <p><span>Lastly, each folder contains a .txt file with the manual annotations of exocytosis events, considered as the ground truth reference. This .txt file is an output of imageJ. The x,y and t coordinates of exocytosis events are given, respectively, by the columns “X”, “Y” and “Slice”. This file reports x and y coordinates in pixels and time as frame number.</span></p> <p><span>Finally, the dataset contains an excel file (.xlsx) giving for each file:</span></p> <p><span>-Folder name</span></p> <p><span>-The total duration of the movie (based on the actual computer saving times of the frames and not the theoretical time set by the microscope)</span></p> <p><span>-The number of frames</span></p> <p><span>-The invert of the frame rate</span></p> <p><span>-The background <em>i.e.</em> average signal in a region outside the cell at t=1</span></p> <p><span>-The signal <em>i.e. </em>average signal in a region inside the cell at t=1</span></p> <p><span>-The classical SBR, computed as the ratio of the two previous columns</span></p> <p><span>-The exocytosis SBR, computed as the ratio between peak intensity of the event (F) and background before the exocytosis event (F<sub>0</sub>) (see paper methods)</span></p> <p><span>-The SBR level, <em>i.e.</em> the classification as low, medium or high SBR based on the exocytosis SBR rank</span></p> <p><span>-Group (either “training” or “inference”) to specify if the movie has been used for ExoDeepFinder training or evaluation</span></p> <p><span> </span></p> <p><strong><span>2\ Robustness Dataset (90 movies)</span></strong></p> <p><span>This dataset contains the data used for the robustness evaluation (figure 2). It contains 4 sub-datasets. Each sub-dataset is made of several folders containing .TIF images corresponding to each frame of the movie, the corresponding .nd file, a .tif cell mask and a .txt file with the manual annotations of exocytosis events, considered as the ground truth reference. This organization is the same as found in the “Training Dataset”. In addition, each sub-dataset contains an excel file (.xlsx), giving for each folder i) the name, ii) number of frames, iii) the invert of the frame and iv) SBR, similarly to the excel file in the “Training Dataset”.</span></p> <p><span> </span></p> <p><strong><span>2.1\ Bafilomycin Dataset (32 movies)</span></strong></p> <p><span>This dataset corresponds to RPE1 cells transfected with VAMP7-pHluorin, seeded on fibronectin-coated coverslips and treated with Bafilomycin A1 (at 100nM for 1h). Each cell is imaged before and 1h after the treatment. The following table shows the pairing.</span></p> <table> <tbody> <tr> <td> <p><strong><span>Before Bafilomycin A1</span></strong></p> </td> <td> <p><strong><span>After Bafilomycin A1</span></strong></p> </td> </tr> <tr> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell1avant</span></p> </td> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell1apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell2avant</span></p> </td> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell2apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell3avant</span></p> </td> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell3apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell4avant</span></p> </td> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell4apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell5avant</span></p> </td> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell5apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell6avant</span></p> </td> <td> <p><span>TIRF 191204 VAMP7 +Bafalomycine A_Cell6apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell1avant</span></p> </td> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell1apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell2avant</span></p> </td> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell2apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell3avant</span></p> </td> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell3apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell4avant</span></p> </td> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell4apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell5avant</span></p> </td> <td> <p><span>TIRF 211118 VAMP7 + Bafilomycine A_Batch1_Cell5apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell1avant</span></p> </td> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell1apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell2avant</span></p> </td> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell2apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell3avant</span></p> </td> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell3apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell4avant</span></p> </td> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell4apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell5vant</span></p> </td> <td> <p><span>TIRF 211223 VAMP7 + Bafilomycin 3_Batch1_Cell5apres</span></p> </td> </tr> </tbody> </table> <p><span> </span></p> <p><strong><span>2.2\ Histamine Dataset (34 movies)</span></strong></p> <p><span>This dataset corresponds to RPE1 cells transfected with VAMP7-pHluorin, seeded on fibronectin-coated coverslips and treated with histamine (at 100µM). Each cell is imaged before and immediately after the treatment. The following table shows the pairing.</span></p> <table> <tbody> <tr> <td> <p><strong><span>Before Histamine</span></strong></p> </td> <td> <p><strong><span>After Histamine</span></strong></p> </td> </tr> <tr> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell1AvantStimulation</span></p> </td> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell1ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell2AvantStimulation</span></p> </td> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell2ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell3ApresStimulation</span></p> </td> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell3Apres2Stimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell4AvantStimulation</span></p> </td> <td> <p><span>TIRF 190516 VAMP7 + Histamine_Cell4ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell1AvantStimulation</span></p> </td> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell1ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell2AvantStimulation</span></p> </td> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell2ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell3AvantStimulation</span></p> </td> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell3ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell4AvantStimulation</span></p> </td> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell4ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell6AvantStimulation</span></p> </td> <td> <p><span>TIRF 190517 VAMP7 + Histamine (2 replicat)_Batch1_Cell6ApresStimulation</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell1avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell1apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell2avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell2apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell3avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell3apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell4avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell4apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell5avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell5apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell6avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell6apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell7avant</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell7apres</span></p> </td> </tr> <tr> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell8avant1</span></p> </td> <td> <p><span>TIRF 220121 VAMP7 + Histamine 3_Batch1_Cell8avant</span></p> </td> </tr> </tbody> </table> <p><span> </span></p> <p><strong><span>2.3\ HeLa Dataset (14 movies)</span></strong></p> <p><span>This dataset corresponds to HeLa cells transfected with VAMP7-pHluorin and seeded on fibronectin-coated coverslips.</span></p> <p><span> </span></p> <p><strong><span>2.4\ CD63 Dataset (10 movies)</span></strong></p> <p><span>This dataset corresponds to RPE1 cells transfected with CD63-pHluorin and seeded on fibronectin-coated coverslips.</span></p> <p><span> </span></p>
Data for: Screening of stapled peptides for inhibition of calcium-triggered exocytosis
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Novel role of TRPM4 ion channel in exocytosis
<p>Under physiological conditions, the widely expressed calcium-activated TRPM4 channel conducts sodium into the cell. This sodium influx depolarizes the plasma membrane and reduces the driving force for calcium entry. Aberrant expression or function of TRPM4 has been reported in various diseases, including different types of cancer. TRPM4 is localized mainly in the plasma membrane, but is also found in intracellular vesicles, which can undergo exocytosis. In this study, we show that calcium-induced exocytosis in the colorectal cancer cell line HCT116 is dependent on TRPM4. In addition, findings from prostate cancer cell lines point to a more general role for TRPM4 in calcium-induced exocytosis in cancer cells. Furthermore, calcium-induced exocytosis depends on TRPM4 ion conductivity. Additionally, an increase in intracellular calcium results in the delivery of TRPM4 to the plasma membrane. This process also depends on TRPM4 ion conductivity. TRPM4-dependent exocytosis and delivery of TRPM4 to the plasma membrane is mediated by SNARE proteins. Finally, we provide evidence that calcium-induced exocytosis depends on TRPM4 ion conductivity not within the plasma membrane, but rather in TRPM4-containing vesicles.</p>
Data from: Fusion pore regulation by cAMP/Epac2 controls cargo release during insulin exocytosis
Regulated exocytosis establishes a narrow fusion pore as initial aqueous connection to the extracellular space, through which small transmitter molecules such as ATP can exit. Co-release of polypeptides and hormones like insulin requires further expansion of the pore. There is evidence that pore expansion is regulated and can fail in diabetes and neurodegenerative disease. Here we report that the cAMP-sensor Epac2 (Rap-GEF4) controls fusion pore behavior by acutely recruiting two pore-restricting proteins, amisyn and dynamin-1, to the exocytosis site in insulin-secreting beta-cells. cAMP elevation restricts and slows fusion pore expansion and peptide release, but not when Epac2 is inactivated pharmacologically or in Epac2-/- (Rapgef4-/-) mice. Consistently, overexpression of Epac2 impedes pore expansion. Widely used antidiabetic drugs (GLP-1 receptor agonists and sulfonylureas) activate this pathway and thereby paradoxically restrict hormone release. We conclude that Epac2/cAMP controls fusion pore expansion and thus the balance of hormone and transmitter release during insulin granule exocytosis.
Data from: Fusion pore regulation by cAMP/Epac2 controls cargo release during insulin exocytosis
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Loss of tetraspanin-7 expression reduces pancreatic beta-cell exocytosis Ca2+ sensitivity but has limited effect on systemic metabolism [human]
GEO Series GSE213730. Homo sapiens. 6 samples. Type: Expression profiling by high throughput sequencing.
Protein palmitoylation and sphingolipid metabolism control regulated exocytosis in cytotoxic lymphocytes
GEO Series GSE307513. Homo sapiens. 7 samples. Type: Other.
Liver lipophagy ameliorates nonalcoholic steatohepatitis through lysosomal lipid exocytosis
GEO Series GSE185911. Mus musculus. 6 samples. Type: Expression profiling by high throughput sequencing.
Paramecium tetraurelia trichocyst exocytosis recovery
GEO Series GSE17930. Paramecium tetraurelia. 12 samples. Type: Expression profiling by genome tiling array.
Galns-deficiency enhances lysosomal exocytosis and alters cartilage development in zebrafish
GEO Series GSE300957. Danio rerio. 8 samples. Type: Expression profiling by high throughput sequencing.
Loss of tetraspanin-7 expression reduces pancreatic beta-cell exocytosis Ca2+ sensitivity but has limited effect on systemic metabolism [mouse]
GEO Series GSE213729. Mus musculus. 4 samples. Type: Expression profiling by high throughput sequencing.
Data from: Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis
Neurotransmitter release is mediated by the fast, calcium-triggered fusion of synaptic vesicles with the presynaptic plasma membrane, followed by endocytosis and recycling of the membrane of synaptic vesicles. While many of the proteins governing these processes are known, their regulation is only beginning to be understood. Here we have applied quantitative phosphoproteomics to identify changes in phosphorylation status of presynaptic proteins in resting and stimulated nerve terminals isolated from the brains of Wistar rats. Using rigorous quantification, we identified 252 phosphosites that are either up- or downregulated upon triggering calcium-dependent exocytosis. Particularly pronounced were regulated changes of phosphosites within protein constituents of the presynaptic active zone, including bassoon, piccolo, and RIM1. Additionally, we have mapped kinases and phosphatases that are activated upon stimulation. Overall, our study provides a snapshot of phosphorylation changes associated with presynaptic activity and provides a foundation for further functional analysis of key phosphosites involved in presynaptic plasticity.
Data from: Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis
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Pancreatic islets communicate with lymphoid tissues via exocytosis of insulin peptides
GEO Series GSE114824. Mus musculus. 8 samples. Type: Expression profiling by high throughput sequencing.
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These curated guides explain access requirements, typical timelines, costs, and reuse considerations for widely used research datasets.
Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.