All-trans retinoic acid induces synaptic plasticity in human cortical neurons
- Department of Neuroanatomy, Institute of Anatomy and Cell Biology, Faculty of Medicine, University of Freiburg, Germany
- Department of Neurosurgery, Medical Center and Faculty of Medicine, University of Freiburg, Germany
- Center for Basics in Neuromodulation (NeuroModulBasics), Faculty of Medicine, University of Freiburg, Germany
- Institute of Clinical Neuroanatomy, Dr. Senckenberg Anatomy, Neuroscience Center, Goethe-University Frankfurt, Germany
- Center Brain Links Brain Tools, University of Freiburg, Germany
Figures
Figure 1 with 1 supplement
All-trans retinoic acid (atRA) induces plasticity of excitatory synapses in human cortical slices.
(A) A representative human cortical slice stained for NeuN. Scale bar = 200 µm. (B) Recorded and post hoc-labeled superficial (layer 2/3) pyramidal neuron. Scale bar = 100 µm. (C) Sample traces of …
Figure 1—figure supplement 1
All-trans retinoic acid (atRA) induces excitatory synaptic strengthening in human superficial (layer 2/3) pyramidal neurons – an in-sample control analysis.
(A) A total amount of eight human neocortical samples were collected for this study, while cortical slices from six samples were included in this dataset. In each sample, acute cortical slices were …
Figure 2 with 1 supplement
All-trans retinoic acid (atRA) induces dendritic spine plasticity in human cortical slices.
(A) Example of dendritic segments of post hoc-labeled superficial (layer 2/3) pyramidal neurons in atRA- (1 µM, 6–10 hr) and vehicle-only-treated slices. Scale bars = 3 µm. (B, C) Group data for …
Figure 2—figure supplement 1
Ultrastructural analysis of excitatory synapses reveals a positive correlation between sizes of the spine head and the spine apparatus organelle.
XY-plots of paired spine head and spine apparatus organelle size (cross section area) were created for both vehicle-only- and atRA-treated human neocortical slices. A positive correlation can be …
Figure 3 with 2 supplements
Effects of all-trans retinoic acid (atRA) in cortical slices prepared from synaptopodin-deficient mice.
(A, B) Group data (A) of AMPA receptor-mediated spontaneous excitatory postsynaptic currents (sEPSCs) recorded from superficial (layer 2/3) pyramidal neurons of the dorsomedial prefrontal cortex in …
Figure 3—figure supplement 1
Analysis of intrinsic cellular properties from superficial pyramidal neurons in wild-type and synaptopodin-deficient slices upon atRA treatment.
(A) Input–output curves of superficial pyramidal neurons and group data of the input resistance and the resting membrane potential in wild-type (Synpo+/+) and synaptopodin-deficient slices (Synpo–/–)…
Figure 3—figure supplement 2
Comparison of baseline spontaneous excitatory synaptic transmission in wild-type, synaptopodin-deficient, and transgenic GFP/Synpo superficial pyramidal neurons of the medial prefrontal cortex.
We found a significant higher tone of both sEPSC amplitude and frequency in synaptopodin-deficient preparations under baseline conditions when compared to wild-type preparations. Of note, no …
Figure 4 with 1 supplement
All-trans retinoic acid (atRA)-induced strengthening of excitatory synapses depends on mRNA translation, but not gene transcription.
(A, B) Group data (A) of AMPA receptor-mediated spontaneous excitatory postsynaptic currents (sEPSCs) recorded from superficial (layer 2/3) pyramidal neurons of the dorsomedial prefrontal cortex in …
Figure 4—figure supplement 1
Intrinsic cellular properties of superficial pyramidal neurons upon atRA treatment and simultaneous pharmacological inhibition of either gene transcription or mRNA translation.
(A–C) Actinomycin D (5 µg/ml) was used to pharmacologically inhibit gene transcription during the atRA treatment. When co-incubated with actinomycin D, atRA did change neither the resting membrane …
Figure 5
Pharmacologic inhibition of mRNA translation prevents all-trans retinoic acid (atRA)-induced synaptic plasticity in human cortical slices.
(A, B) Group data (A) of AMPA receptor-mediated spontaneous excitatory postsynaptic currents (sEPSCs) and cumulative distribution (B) of sEPSC amplitudes recorded from superficial (layer 2/3) …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Anti-Synaptopodin (Rabbit polyclonal) | Synaptic Systems | Cat#: 163002 RRID:AB_887825 | IF (‘1:1000’) EM (‘1:100’) |
| Antibody | Anti-NeuN (Rabbit polyclonal) | Abcam | Cat#: ab104225 RRID:AB_10711153 | IF (‘1:500’) |
| Antibody | Anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Goat polyclonal) | Invitrogen | Cat#: A-11034, RRID:AB_2576217 | IF (‘1:1000’) |
| Antibody | Anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 555 (Goat polyclonal) | Invitrogen | Cat#: A-32732, RRID:AB_2633281 | IF (‘1:1000’) |
| Antibody | Anti-Rabbit IgG Nanogold-Fab' (goat polyclonal) | Nanoprobes | Cat#: 2004 RRID:AB_2631182 | EM (‘1:100’) |
| Biological sample (Homo sapiens), male and female | Sample | Biobank of the Department for Neurosurgery at the Faculty of Medicine, University of Freiburg, AZ 472/15_160880 | Approval of the Local Ethics Committee, University of Freiburg, AZ 593/19 | |
| Chemical compound, drug | DAPI (1 mg/ml in water) | Thermo Scientific | Cat#: 62248 | IF and post hoc labeling (‘1:5000’) |
| Chemical compound, drug | Pierce16% Formaldehyde (w/v), methanol-free | Thermo Scientific | Cat#: 28906 | Final concentration: (4% in PBS) |
| Chemical compound, drug | Glutardialdehyd | Carl Roth | Cat#: 4157.2 | Final concentration: 2.5% (TEM) and 0.1% (Immunogold) |
| Chemical compound, drug | All-trans retinoic acid | Sigma–Aldrich | Cat#: R2625 | Final concentration: 1 µM |
| Chemical compound, drug | Anisomycin | Abcam | Cat#: ab120495 | Final concentration: 10 µM |
| Chemical compound, drug | Actinomycin D | Sigma–Aldrich | Cat#: A9415 | Final concentration: 5 µg/ml |
| Commercial assay, kit | HQ Silver Enhancement Kit | Nanoprobes | Cat#: 2012 | |
| Genetic reagent Mus musculus, male | B6.129-Synpotm1Mndl/Dllr; Synpo−/− | Vlachos et al., 2013 PMID:23630268 | MGI: 6423115 | Obtained from Deller Lab (Frankfurt) |
| Genetic reagent Mus musculus, male | B6.Cg-Synpotm1MndlTg(Thy1-Synpo/GFP)1Dllr/Dllr; Thy1-GFP/Synpo+/− x Synpo−/− | Vlachos et al., 2013 PMID:23630268 | MGI: 6423116 | Obtained from Deller Lab (Frankfurt) |
| Peptide, recombinant protein | Streptavidin, Alexa Fluor 488-Conjugate | Invitrogen | Cat#: S32354 RRID:AB_2315383 | Post hoc labeling (‘1:1000’) |
| Software, algorithm | Prism | GraphPad | RRID:SCR_002798 | |
| Software, algorithm | Clampfit (pClamp software package) | Molecular Devices | RRID:SCR_011323 | |
| Software, algorithm | ImageJ | RRID:SCR_003070 | ||
| Software, algorithm | Photoshop | Adobe | RRID:SCR_014199 | |
| Strain, strain background Mus musculus | C57BL/6J; Synpo+/+ | Jackson Laboratory | RRID: IMSR_JAX:000664 |
Additional files
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Supplementary file 1
Cortical resection samples.
- https://cdn.elifesciences.org/articles/63026/elife-63026-supp1-v1.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/63026/elife-63026-transrepform-v1.pdf
