Structural plasticity of dendritic secretory compartments during LTP-induced synaptogenesis
- The University of Texas at Austin, United States
Figures
Figure 1
Within-slice experimental design and electrophysiological outcome.
(A) Illustration of an acute slice from a P15 rat hippocampus with a recording electrode (rec.) in the middle of CA1 stratum radiatum between two bipolar stimulating electrodes (S1 and S2). S1 and …
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Figure 1—source data 1
Excel spreadsheet containing the raw numbers that generated the graphs and waveforms for these experiments.
- https://doi.org/10.7554/eLife.46356.003
Figure 2 with 1 supplement
The limited occupancy of spines by SER does not increase during spinogenesis in the LTP condition.
(A) Sample serial section EMs (left) and representative 3D reconstructions of dendrites (right) from control (top) and LTP (bottom) conditions, illustrating dendrites (yellow), SER (green), and …
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Figure 2—source data 1
Excel spreadsheets containing the raw numbers that generated the graphs in each part of this figure along with the summary of statistics.
- https://doi.org/10.7554/eLife.46356.006
Figure 2—figure supplement 1
All analyzed dendrites fully reconstructed with SER, aligned left to right from least to greatest spine density.
Scale cube is 0.5 µm on each side.
Figure 3
Reduction in shaft SER following LTP.
(A) Electron micrographs showing the dendrite (yellow), SER (green), and synapses (red). For both control and LTP, the SER in the aspiny segments forms small cross-sectioned tubules, whereas in the …
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Figure 3—source data 1
Excel spreadsheets containing the raw numbers that generated the graphs in each part of this figure along with the summary of statistics.
- https://doi.org/10.7554/eLife.46356.008
Figure 4 with 9 supplements
Identification of endosomal compartments.
(A) Model of the dendritic endosomal pathway. Clathrin-coated pits (CPs) invaginate, becoming clathrin-coated vesicles (CVs) and large vesicles (LVs) after coat shedding. Large vesicles fuse to form …
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Figure 4—source data 1
Excel spreadsheets containing details of the locations of each object in Figure 4.
- https://doi.org/10.7554/eLife.46356.016
Figure 4—figure supplement 1
Sample images from the LTP condition of dendritic (yellow) recycling complex with multiple tubules (pink) entering the spine neck, and two small vesicles (purple arrow) in a different dendritic spine.
D28 FZYJV sections 108–111. Scale bar 0.5 µm.
Figure 4—figure supplement 2
Sample images of coated pit (orange) inside the dendritic shaft (yellow) from a dendrite in the LTP condition, D25 DCPBM sections 121–124.
Scale bar 0.5 µm.
Figure 4—figure supplement 3
Sample images of sorting complex (turquoise) inside the dendritic shaft from the control condition, with one tubule entering a spine neck (right side row 3).
D26 PWCNZ sections 41–44. Scale bar 0.5 µm.
Figure 4—figure supplement 4
Sample image of an amorphous vesicle in the dendritic shaft of the LTP condition from D35 DCPBM sections 25–28.
Scale bar 0.5 µm.
Figure 4—figure supplement 5
Sample images of degradative lysosome (black) in the dendritic shaft (yellow) of the LTP condition from D17 FZYJV sections 146–149.
Scale bar 0.5 µm.
Figure 4—figure supplement 6
Sample images of degradative whorl (black) in a dendrite (yellow) of the control condition from D69 FXBVK sections 176–180.
Scale bar 0.5 µm.
Figure 4—video 1
Video paging through dendritic from the LTP condition including sections 96–121 of D28 FZYJV.
https://doi.org/10.7554/eLife.46356.017
Figure 4—video 2
A sorting complex (turquoise) in a dendrite (yellow) from the control condition is D26 PWCNZ and includes sections 35–46.
https://doi.org/10.7554/eLife.46356.018
Figure 4—video 3
A degradative whorl (black) in a dendrite of the control condition from D69 FXBVK sections 170–187.
https://doi.org/10.7554/eLife.46356.019
Figure 5 with 2 supplements
Increased occurrence of endosomes in small spines after LTP.
(A) Sample serial EM sections and representative 3D reconstructed dendrites illustrate the distribution of endosomal compartments from control and LTP conditions. Dendrites are yellow, synapses are …
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Figure 5—source data 1
Excel spreadsheets containing the raw numbers that generated the graphs in each part of this figure along with the summary of statistics.
- https://doi.org/10.7554/eLife.46356.023
Figure 5—figure supplement 1
All analyzed dendrites fully reconstructed with constructive endosomes, aligned left to right from least to greatest spine density.
Scale cube is 0.5 µm on each side.
Figure 5—figure supplement 2
All analyzed dendrites fully reconstructed with intracellular degradative structures, aligned left to right from least to greatest spine density.
Scale cube is 0.5 µm on each side.
Figure 6
Model of the contribution of dendritic secretory compartments to LTP-induced synaptogenesis.
Smooth endoplasmic reticulum (SER, green), postsynaptic density (PSD, red), small vesicle or recycling endosome (RE, turquoise), new silent spines (orange), control activation (Con), theta-burst …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Rattus norvegicus, male) | Long-Evans Rat | Charles River | Charles River strain# 006; RRID:RGD_2308852 | |
| Chemical compound, drug | Potassium ferrocyanide | Sigma-Aldrich | Cat# P3289 | |
| Chemical compound, drug | Osmium tetroxide | Electron Microscopy Sciences | Cat# 19190 | |
| Chemical compound, drug | Uranyl acetate | Electron Microscopy Sciences | Cat# 22400 | |
| Chemical compound, drug | LX-112 embedding kit | Ladd Research Industries | Cat# 21210 | |
| Chemical compound, drug | Lead nitrate | Ladd Research Industries | Cat# 23603 | |
| Chemical compound, drug | Pioloform F | Ted Pella | Cat# 19244 | |
| Software, algorithm | Igor Pro 4 | WaveMetrics | https://www.wavemetrics.net/ | |
| Software, algorithm | Reconstruct | Fiala, 2005 | Executable and manual: http://synapseweb.clm.utexas.edu/software-0 | Source at:https://github.com/orgs/SynapseWeb/teams/reconstruct-developers |
| Software, algorithm | STATISTICA 13 Academic | Tibco | https://onthehub.com//statistica/ | |
| Other | Tissue slicer | Stoelting | Cat # 51425 | |
| Other | Vibratome | Leica Biosystems | VT1000S | |
| Other | Ultramicrotome | Leica Biosystems | UC6 | Used with a Diatome Ultra35 knife |
| Other | SynapTek Grids | Ted Pella | Cat# 4514 or 4516 | |
| Other | Diffraction grating replica | Electron Microscopy Sciences | Cat# 80051 | |
| Other | Transmission electron microscope | JEOL | JEM-1230 | |
| Other | Harris Lab wiki | Harris Lab | https://wikis.utexas.edu/display/khlab/ | This wiki site hosts experimental methods used for this paper and updates. |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.46356.025
