Interneuron FGF13 regulates seizure susceptibility via a sodium channel-independent mechanism
- Cardiovascular Research Institute, Weill Cornell Medicine, United States
- Department of Neurological Surgery and Brain and Mind Research Institute, Weill Cornell Medicine of Cornell University, New York Presbyterian Hospital, United States
- Department of Pediatrics, Division of Pediatric Neurology, Weill Cornell Medicine, United States
- Brain and Mind Research Institute, Weill Cornell Medicine, United States
Figures
Figure 1 with 1 supplement
Whole brain knockout of Fgf13 results in premature death and seizure susceptibility.
(A) Breeding scheme to generate Nestin-Fgf13 cKO, Nestin-Fgf13 Het, and wildtype littermates. (B) Western blots of whole brain Nestin-Fgf13 cKO (KO) and wildtype (WT) littermates at P2 validates Fgf1…
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Figure 1—source data 1
Original TIFF files saved from BioRad Gel doc system for gels shown in Figure 1B, together with a PDF file identifying the respective portions displayed.Figure 1B.
- https://cdn.elifesciences.org/articles/98661/elife-98661-fig1-data1-v1.zip
Figure 1—figure supplement 1
Fgf13 splice variants are differentially expressed in hippocampal cell types.
(A) Fgf13 generates five alternatively spliced variants. (B) Knockout of in Fgf13 in Nestin neurons eliminates all isoforms, while knockout of Fgf13 in Emx excitatory neurons eliminates the S and U …
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Figure 1—figure supplement 1—source data 1
Original TIFF files saved from BioRad Gel doc system for gels shown in Figure 1—figure supplement 1B, together with a PDF file identifying the respective portions displayed.
- https://cdn.elifesciences.org/articles/98661/elife-98661-fig1-figsupp1-data1-v1.zip
Figure 2 with 2 supplements
Excitatory neuronal knockout of Fgf13 does not result in premature death and seizure susceptibility.
(A) Western blot shows partial loss of Fgf13 from Emx1-Fgf13 cKO hippocampal tissue, compared to full knockout in Nestin-Fgf13 cKO hippocampus. Vinculin used as a loading control. (B) Fluorescent …
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Figure 2—source data 1
Original TIFF files saved from Bio-Rad Gel doc system for gels shown in Figure 2A, together with a PDF file identifying the respective portions displayed.
- https://cdn.elifesciences.org/articles/98661/elife-98661-fig2-data1-v1.zip
Figure 2—figure supplement 1
Excitatory neuronal knockout of Fgf13 results in loss of FGF13-S.
Western blot performed with FGF13-S-specific antibody. Vinculin used as a loading control.
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Figure 2—figure supplement 1—source data 1
Original TIFF files saved from Bio-Rad Gel doc system for gels shown in Figure 2—figure supplement 1, together with a PDF file identifying the respective portions displayed.
- https://cdn.elifesciences.org/articles/98661/elife-98661-fig2-figsupp1-data1-v1.zip
Figure 2—figure supplement 2
Excitatory neuronal knockout of Fgf13 does not result in sodium current deficits.
(A) Primary hippocampal neuron cultures were generated from Emx1-Cre wildtype (left) or Emx1-Fgf13 cKO male mice, and Emx1 +excitatory cells were labeled by infection with a Cre-dependent …
Figure 3 with 4 supplements
Inhibitory neuronal knockout of Fgf13 recapitulates premature death and seizure susceptibility.
(A) Quantification of hippocampal interneuron histology reveals 31% of Gad2+ interneurons co-express FGF13. (B) Fluorescent immunohistochemistry of hippocampal tissue validates Fgf13 knockout in …
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Figure 3—source data 1
Original TIFF files saved from Bio-Rad Gel doc system for gels shown in Figure 3C and PDF, together with a identifying the respective portions displayed.
- https://cdn.elifesciences.org/articles/98661/elife-98661-fig3-data1-v1.zip
Figure 3—figure supplement 1
Characterization of FGF13 abundance in hippocampus of Gad2-Fgf13 cKO mice.
(A) Fluorescent immunohistochemistry with a pan-FGF13 antibody on hippocampal tissue in Gad2-GFP reporter mice. (B) Immunohistochemistry of hippocampal tissue in Gad2-GFP-Fgf13 cKO shows loss of …
Figure 3—figure supplement 2
Fgf13 splice variants are differentially expressed in hippocampal cell types.
(A)Fgf13-S and -VY exhibit distinct expression in the hippocampus of wildtype and cell-type-specific knockouts. (B) Example images showing decreased Fgf13-S expression from Nestin-Fgf13 cKO and …
Figure 3—figure supplement 3
An unbiased metabolomic analysis on whole brain lysate from Gad2-Fgf13 cKO and wildtype mice reveals marked differences between genotypes.
(A) PCA plot showing three WT and three Gad2-Fgf13 cKO mice. (B) Volcano plot of measured metabolites. Colored data show metabolites with p<0.05 and Log2FC >1 or Log2FC <1 (KO v WT). N=3 mice each.
Figure 3—figure supplement 4
Knockout of Fgf13 from MGE-derived interneurons recapitulates premature death and seizure susceptibility.
(A) Re-analysis of single-cell RNA-seq data (1) reveals that Fgf13 is expressed in MGE-derived interneurons such as the axo-axonic and MGE-derived neurogliaform cell types, as well as CGE-derived …
Figure 4
Inhibitory neuronal knockout of Fgf13 results in deficits of synaptic transmission.
(A) Example traces from spontaneous excitatory postsynaptic currents (sEPSCs). (B) Cumulative fraction (Kolmogorov Smirnov test, ****, p<0.0001) frequency, and current amplitude for sEPSCs (t-test, …
Figure 5 with 1 supplement
Inhibitory neuronal knockout of Fgf13 results in deficits of interneuron excitability.
(A) Examples of FGF13-stained neurons from primary hippocampal neuron cultures generated from Gad2-Cre wildtype (top) and Gad2-Fgf13 cKO (bottom) male mice (scale bar, 20 μm). (B) Example traces of …
Figure 5—figure supplement 1
Inhibitory neuronal knockout of Fgf13 does not result in sodium current deficits.
(A) Wildtype and Gad2-Fgf13 cKO interneurons have no significant difference in APD50, peak voltage amplitude, and dV/dt max values for the first action potential in each spike train. (B) Wildtype …
Figure 6
AAV-mediated expression of FGF13 isoforms rescues excitability deficits in Gad2-Fgf13 cKO neurons.
(A) Examples of FGF13-stained neurons from primary hippocampal neuron cultures generated from Gad2-Fgf13 cKO male mice transduced with AAV8-DIO-GFP only, AAV8-DIO-GFP and AAV8-DIO-Fgf13-S, or …
Videos
Video 1
Nestin-Fgf13 cKO male mice suffered spontaneous seizures in their home cage.
Example spontaneous seizure activity is shown.
Tables
Table 1
Mutant mice are born in Mendelian ratios.
| Mouse strain | Probability density function |
|---|---|
| Nestin-Fgf13 | 0.23 |
| Gad2-Fgf13 | 0.22 |
| Nkx2.1-Fgf13 | 0.24 |
| Emx1-Fgf13 | 0.21 |
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Mutant mice were born in expected Mendelian ratios. Despite embryonic targeted deletion of Fgf13, mutant suffered no prenatal mortality related to developmental deficits.
Additional files
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Supplementary file 1
Primers used for real time quantitative polymerase chain reaction.
Forward and reverse primer pair sequences and, where relevant, the PrimerBank ID.
- https://cdn.elifesciences.org/articles/98661/elife-98661-supp1-v1.docx
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Supplementary file 2
Metabolite set enrichment analysis in Gad2-Fgf13 cKO vs. WT.
Unbiased metabolites identified by mass spectometery in Gad2-Fgf13 cKO vs. WT.
- https://cdn.elifesciences.org/articles/98661/elife-98661-supp2-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/98661/elife-98661-mdarchecklist1-v1.docx
