Role of matrix metalloproteinase-9 in neurodevelopmental deficits and experience-dependent plasticity in Xenopus laevis
- Department of Neuroscience, Brown University, United States
- The Scripps Research Institute, United States
Figures
Figure 1
MMP-9 overexpression increases spontaneous synaptic activity and network connectivity.
(A) Representative sEPSCs and sIPSCs recorded at −45 mV and +5 mV, respectively, from control and MMP-9 transfected cells. (B) Boxplots showing frequency of sEPSCs and sIPSCs for GFP control, MMP-9 …
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Figure 1—source data 1
MMP9 overexpression data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig1-data1-v3.xlsx
Figure 2 with 1 supplement
Pharmacological inhibition of MMP-9 rescues VPA-induced effects on synaptic transmission.
(A) Representative sEPSCs (left) and evoked response (right) form control, VPA and VPA + SB-3CT-treated animals. (B) Boxplots showing frequency of sEPSCs for control, VPA, VPA + SB-3CT, and SB-3CT …
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Figure 2—source data 1
SB3CT source data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig2-data1-v3.xlsx
Figure 2—figure supplement 1
Exposure to VPA results in increased synapse density.
(A) Box plot showing synapse density for control and VPA groups. VPA exposed tadpoles showed increased synapse density compared to control group. (B) Representative image from control animal …
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Figure 2—figure supplement 1—source data 1
PSD95 overexression source data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig2-figsupp1-data1-v3.xlsx
Figure 3
Morpholino targeted against MMP-9 rescues VPA-induced effects.
(A) Western blot image (left) indicating MMP-9 levels (75kD band) in control (scrambled) MO and MMP-9 MO transfected groups. Boxplot (right) shows relative MMP-9 levels for control MO and MMP MO …
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Figure 3—source data 1
Morpholino source data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig3-data1-v3.xlsx
Figure 4
Inhibition of MMP-9 reverses VPA-induced behavioral effects.
(A) Boxplots showing seizure frequencies for Control, VPA and VPA + SB-3CT groups. Seizure frequency is increased in the VPA group, but this effect is rescued by addition of SB-3CT. Data are …
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Figure 4—source data 1
Behavior data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig4-data1-v3.xlsx
Figure 5 with 1 supplement
Enhanced visual activity increases MMP-9 levels and MMP-9 inhibition prevents visual-activity induced dendritic growth.
(A) Western blot analysis specifying MMP-9 levels (75 kDa band) after lightbox, dark exposure and naive controls (normal). (B) Box plot showing relative MMP-9 levels for light box treatment …
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Figure 5—source data 1
Visual stimulation source data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig5-data1-v3.xlsx
Figure 5—figure supplement 1
Exposure to VPA results in significantly less WFA staining intensity in comparison to control animals.
(A) Schematic representation of the tadpole nervous system showing hindbrain (HB), optic tectum (OT) and olfactory bulb (OB). Optic tectum consists of a well-defined layer of cell bodies (yellow) …
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Figure 5—figure supplement 1—source data 1
WFA staining data.
- https://cdn.elifesciences.org/articles/62147/elife-62147-fig5-figsupp1-data1-v3.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Xenopus laevis) | Tadpole | Nasco | WT and Albino strains; LM00456, LM00456(A) | Bred in house |
| Chemical compound, drug | Valproic Acid | Sigma | Cat. No. P4543 | Drug |
| Chemical compound, drug | SB-3CT | Tocris | Cat. No. 6088 | MMP9/2 inhibitor |
| Recombinant DNA reagent | pCMV-SPORT6, MMP-9, GFP | Invitrogen | pCMV-SPORT6 (backbone) | Expressing Xenopus MMP9 and GFP, or GFP alone, or MMP9-FLAG |
| Genetic reagent | MMP-9 MO | GeneTools Bestman and Cline, 2014 | DOI:10.1007/978-1-62703-655-9_11 | Morpholino antisense oligonucleotide- lissamine tagged, Used to drive down expression of MMP9 |
| Genetic reagent | Control MO | GeneTools Bestman and Cline, 2014 | DOI:10.1007/978-1-62703-655-9_11 | Used as a transfection control, Morpholino antisense oligonucleotide- lissamine tagged |
| Antibody | Anti-MMP9 (Rabbit polyclonal) | EMD Millipore | Ab19016; RRID:AB_91090 | WB (1:2000) |
| Antibody | Anti-Rabbit-HRP (goat polyclonal) | BioRad | #1662408EDU; RRID:AB_11125345 | Secondary Ab for WB |
| Commercial assay or kit | Pierce ECL-western blot substrate | Thermo Fisher | Cat#32209 | For visualizing labeling |
| Recombinant DNA reagent | pCALNL-GFP (plasmid) | Schohl et al., 2020 , DOI: 10.3389/fncir.2020.00047 | For single cell labeling | |
| Recombinant DNA reagent | pCAG-Cre (plasmid) | Schohl et al., 2020, DOI: 10.3389/fncir.2020.00047 | For single cell labeling | |
| Recombinant DNA reagent | pBE-PSD-95 GFP | Sanchez et al., 2006; DOI: 10.1242/dev.02409 | For synaptic labeling | |
| Recombinant DNA reagent | pCMV-mCherry | Schwartz et al., 2009; DOI:10.1016/j.neuron.2011.02.055 | Cell filling mCherry | |
| Chemical compound, drug | F-WFA | Vector Labs | FL-1351–2 | Fluorescein-labeled Wisteria floribunda agglutinin, 1:500 dilution for staining PNNs |
Additional files
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Supplementary file 1
Intrinsic cellular properties in different experimental groups.
Input resistance and membrane capacitance values showed no significant differences between experimental groups.
- https://cdn.elifesciences.org/articles/62147/elife-62147-supp1-v3.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/62147/elife-62147-transrepform-v3.docx
