Figures and data in Inhibitory CCK+ basket synapse defects in mouse models of dystroglycanopathy

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7 figures, 3 tables and 3 additional files

Figures

Figure 1 with 2 supplements

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Dystroglycan synaptic localization and glycosylation in mouse models of dystroglycanopathy.

(A) Schematic depiction of Dystroglycan in different mouse models. The IIH6 antibody recognizes the matriglycan repeats on extracellular αDag1. Hippocampal CA1 of P30 mice immunostained for Dystroglycan glycosylation (IIH6, green) and nuclear marker Hoechst (blue) show puncta of glycosylated Dystroglycan localized to the perisomatic region of pyramidal cells and to blood vessels (magenta arrowheads); scale bar = 50 μm. Lower panels show cell bodies in SP; scale bar = 25 μm. CA1 layers: SO, *stratum oriens*; SP, *stratum pyramidale*; SR, *stratum radiatum*. (B) WGA-enriched lysates from P0 forebrain were immunoblotted for IIH6, β-Dag1, and β-tubulin. (**C–D**) Quantification of immunoblot in (B). Error bars show mean + SEM. See Supplementary file 1 for Ns.

Figure 1—source data 1 Raw data for quantification in Figure 1C, D.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig1-data1-v1.csv
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Figure 1—figure supplement 1

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*Emx1^Cre* drives recombination in forebrain excitatory neurons and astrocytes, but not interneurons or microglia.

(A) Left; endogenous red fluorescence in the forebrain of P60 *Emx1^Cre:R26^{LSL-H2B-mCherry}* reporter mice (scale bar = 1 mm). Right; coronal section of the forebrain from *Emx1^Cre:R26^{LSL-H2B-mCherry}* mice showing robust nuclear mCherry signal (magenta) in the cortex and hippocampus (scale bar = 500 μm). (**B–E**) mCherry⁺ nuclei shown with markers of multiple cell types in the brain including astrocytes (GFAP, green) (B), microglia (Iba1, green) (C), and neurons (NeuN, green) (D) (scale bar = 100 μm). Insets show enlarged images of mCherry⁺ nuclei with cell type markers (scale bar = 20 μm). (E) mCherry⁺ nuclei with markers for different interneuron subtype markers (yellow arrowheads) (scale bar = 100 μm) (VIP = Vasoactive intestinal peptide, NECAB = Neuronal calcium-binding protein 1, SOM = Somatostatin). *CA1 layers: SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 1—figure supplement 2

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Dystroglycan glycosylation is required for synaptic localization.

(A) Immunostaining for matriglycan (IIH6, green) and β-Dag1 (magenta) in CA1 pyramidal cells of WT controls, *Emx1^Cre:Dag1* cKOs, *Emx1^Cre:Pomt2* cKOs, and *Dag1^cyto/-* mutants. Scale bars = 25 μm. (B) WGA-enriched lysates from P21 to P30 hippocampus were immunoblotted for IIH6 and β-Dag1. (**C–D**) Quantification of immunoblot in (B). Error bars show mean + SEM. See Supplementary file 1 for Ns.

Figure 1—figure supplement 2—source data 1 Raw data for quantification in Figure 1—figure supplement 2C, D.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig1-figsupp2-data1-v1.csv
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Figure 2 with 2 supplements

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Dystroglycan is required for cortical neuron migration in a glycosylation-dependent manner and independent of intracellular interactions.

Immunostaining for cortical layer markers (A) Cux1 (layers 2–4) and (B) Tbr1 (layers 3 and 6) in P30 somatosensory cortex (scale bar = 200 μm). Layer markers are shown in green. Nuclear marker Hoechst is shown in magenta. Quantification of fluorescence intensity of layer markers shown for (C) Cux1 and (D) Tbr1. Shaded regions of intensity profile illustrate ± SEM. See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. CC, corpus callosum; A.U., arbitrary units*.

Figure 2—source data 1 Raw data for quantification in Figure 2C.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig2-data1-v1.csv
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Figure 2—source data 2 Raw data for quantification in Figure 2D.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig2-data2-v1.csv
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Figure 2—figure supplement 1

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Cortical migration is disrupted at midline of *B4gat1^M155T/M155T* but not *Fkrp^P448L/P448L* mutants.

(A) Immunostaining for cortical layer marker Cux1 (layers 2–4) in P30 somatosensory cortex (scale bar = 500 μm). Cux1 is shown in green. Nuclear marker Hoechst is shown in magenta. Cortical migration is normal in both *B4gat1* and *Fkrp* point mutants except at midline, where the *B4gat1* mutants show a migration phenotype not seen in the *Fkrp* point mutants. This phenotype was observed with 100% penetrance in the *B4gat1* mutants and 0% in the *Fkrp* mutants.

Figure 2—figure supplement 2

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Laminin immunoreactivity in adult neocortex appears discontinuous in *Dag1* mutants.

(**A–C**) Immunostaining for ECM protein Laminin (green). Nuclear marker Hoechst is shown in blue. Scale bar = 1000 μm.

Figure 3 with 4 supplements

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*Dag1* is required for CCK⁺/CB₁R⁺ basket IN perisomatic axon targeting in *stratum pyramidale* of hippocampal CA1-3.

(A) Nuclear marker Hoechst (upper panels) shows hippocampal morphology. Granule cell migration is disrupted in dentate gyrus of *Emx1^Cre:Dag1* cKOs and *Emx1^Cre:Pomt2* cKOs (yellow arrows). CA1-3 gross morphology is normal in all models. CB₁R immunostaining (lower panels) shows abnormal CCK⁺/CB₁R⁺ basket interneuron targeting in CA1-3 to varying degrees across models (scale bar = 400 μm). (B) Higher magnification view of CB₁R immunostaining in CA1 (scale bar = 50 μm). (C) Quantification of CA1 CB₁R fluorescence intensity profile. Shaded regions of intensity profile illustrate ± SEM. Gray region highlights SP. See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. A.U., arbitrary units; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 3—source data 1 Raw data for quantification in Figure 3C.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig3-data1-v1.csv
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Figure 3—figure supplement 1

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CCK⁺/CB₁R⁺ IN axon targeting phenotypes in hippocampal CA1 of various *Dag1* cKOs.

CB₁R immunostaining in hippocampal CA1 of control mice (left panels) and *Dag1* cKOs (right panels) generated using *Emx1^Cre* (forebrain progenitors, E10.5) (A), *Nestin^Cre* (neural stem cells in the central and peripheral nervous systems, E11.5) (B), and *NeuroD6^Cre* (forebrain postmitotic excitatory neurons, E12.5) (C). Scale bar = 50 μm. *SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 3—figure supplement 2

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CCK⁺/CB₁R⁺ IN cell numbers are unchanged in *Emx1^Cre:Dag1* cKOs.

(A) Immunostaining in hippocampal CA1 for NECAB1 (left) and PV (right) in *Emx1^Cre:Dag1* control and *Emx1^Cre:Dag1* cKO mice. NECAB1 labeling is evident in PV⁺ cells (example shown in blue inset) and in an additional PV^- population that contains the CCK⁺/CB₁R⁺ population (example shown in magenta inset). The average number of NECAB1⁺;PV^- cells per 100 μm of SP is quantified in (B). Immunostaining for NECAB2 (C) and PV (D) with cell number quantification in (E) and (F). Scale bar = 100 μm. Error bars show mean + SEM. See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. SP, stratum pyramidale; PV, parvalbumin*.

Figure 3—figure supplement 2—source data 1 Raw data for quantification in Figure 3—figure supplement 2B, E, and F.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig3-figsupp2-data1-v1.csv
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Figure 3—figure supplement 3

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Parvalbumin⁺ basket INs do not require *Dag1* for proper axon targeting in hippocampal CA1.

(A) Parvalbumin (PV) immunostaining in P30 hippocampal CA1 shows that PV⁺ basket interneurons exhibit normal distribution in CA1 for all genetic models except for an increase in PV intensity within *stratum pyramidale* of *Emx1^Cre:Dag1* cKOs (scale bar = 50 μm). (B) Quantification of CA1 PV fluorescence intensity profile. Shaded regions of intensity profile illustrate ± SEM. Gray region highlights SP. See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. A.U., arbitrary units; SO, stratum oriens.*

Figure 3—figure supplement 3—source data 1 Raw data for quantification in Figure 3—figure supplement 3B.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig3-figsupp3-data1-v1.csv
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Figure 3—figure supplement 4

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Altered CB₁R expression in cortex and basolateral amygdala of *Dag1* and *POMT2* mutants.

(A) CB₁R immunostaining in P30 coronal sections showing somatosensory cortex (Ctx) and CA1 of hippocampus (Hipp) (scale bar = 400 μm). (B) CB₁R immunostaining (green) shown with nuclear marker Hoechst (magenta) in basolateral amygdala (BLA) (scale bar = 250 μm).

Figure 4

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*Dag1* is required for CCK⁺/CB₁R⁺ IN axon targeting during early postnatal development.

(A) Immunostaining for CB₁R⁺ axon terminals (green) in the hippocampus of *Emx1^Cre:Dag1* controls (left) and cKOs (right) at ages P5-P30. Nuclear marker Hoechst is shown in magenta. White arrowheads indicate migration errors in dentate granule cells *Emx1^Cre:Dag1* cKO mice. Scale bar = 500 μm. (B) Higher magnification images of CB₁R⁺ axon terminals in CA1 of *Emx1^Cre:Dag1* controls (left) and cKOs (right) at ages P5-P30. Scale bar = 100 μm. (C) Quantification of CB₁R fluorescence intensity profile in CA1. Shaded regions of intensity profile illustrate ± SEM. Gray region highlights SP. See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. A.U., arbitrary units; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 4—source data 1 Raw data for quantification in Figure 4C.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig4-data1-v1.csv
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Figure 5 with 3 supplements

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*Dag1* and *Pomt2* cKOs exhibit impaired CB1R⁺ basket synapse formation in *stratum pyramidale* of hippocampal CA1.

P30 coronal sections immunostained for (A) CB₁R and (B) VGAT in hippocampal CA1; merged image in (C) shows CB₁R in magenta and VGAT in green. (**A–C**) Scale bar = 50 μm. (Higher magnification view of SP in (C’); scale bar = 10 μm.) (D) Quantification of VGAT puncta density in SP expressed as a percent of control. (E) Quantification of CB₁R puncta density in SP expressed as a percent of control. (F) Quantification of co-localization between VGAT and CB₁R in SP to estimate putative CB₁R⁺ basket cell synapse formation. Error bars show mean + SEM. (For quantification of puncta densities and co-localization in SO and SR see Figure 5—figure supplement 1.) See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 5—source data 1 Raw data for quantification in Figure 5D, E.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig5-data1-v1.csv
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Figure 5—source data 2 Raw data for quantification in Figure 5F.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig5-data2-v1.csv
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Figure 5—figure supplement 1

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Extended quantification of images in Figure 5A–C.

Quantification within hippocampal CA1 SO and SR regions and all regions pooled together. (A) Quantification of VGAT puncta density expressed as a percent of control. (B) Quantification of CB₁R puncta density expressed as a percent of control. (C) Quantification of co-localization between VGAT and CB₁R to estimate putative CB₁R⁺ basket cell synapse formation. Error bars show mean + SEM. (To see quantification of puncta densities and co-localization in SP see Figure 5D–F.) See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05*.

Figure 5—figure supplement 2

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*Dag1* and *Pomt2* cKOs exhibit increased PV⁺ basket synapse formation in *stratum pyramidale* of hippocampal CA1.

P30 coronal sections immunostained for (A) PV and (B) VGAT in hippocampal CA1; merged image in (C) shows PV in magenta and VGAT in green. Yellow arrows indicate PV⁺ interneuron cell bodies. (**A–C**) Scale bar = 50 μm. (Higher magnification view of SP in (C’); scale bar = 10 μm.) (D) Quantification of PV puncta density in SP expressed as a percent of control. (E) Quantification of co-localization between VGAT and PV in SP to estimate putative PV⁺ basket synapse formation. Error bars show mean + SEM. (To see quantification of puncta densities and co-localization in SO and SR see Figure 5—figure supplement 3.) See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS = p ≥ 0.05. PV, parvalbumin; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 5—figure supplement 2—source data 1 Raw data for quantification in Figure 5—figure supplement 2D.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig5-figsupp2-data1-v1.csv
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Figure 5—figure supplement 2—source data 2 Raw data for quantification in Figure 5—figure supplement 2E.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig5-figsupp2-data2-v1.csv
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Figure 5—figure supplement 3

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Extended quantification of images in Figure 5—figure supplement 2A–C.

Quantification within hippocampal CA1 SO and SR regions and all regions pooled together. (A) Quantification of PV puncta density expressed as a percent of control. (B) Quantification of co-localization between VGAT and PV to estimate putative PV⁺ basket cell synapse formation. Error bars show mean + SEM. (To see quantification of puncta densities and co-localization in SP see Figure 5—figure supplement 2D, E). See Supplementary file 1 for Ns. *Significance: *=*p *< 0.05, **=*p *< 0.01, ***=*p *< 0.001, NS =* p *≥ 0.05. PV, parvalbumin; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum*.

Figure 6 with 1 supplement

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*Dag1* is required for CCK⁺/CB₁R⁺ IN synapse function in hippocampal CA1 in a manner dependent on both glycosylation and intracellular interactions.

(A) Representative traces showing + seconds of sIPSC recordings at baseline (top) and after the addition of carbachol (bottom). (B) Quantification of average sIPSC frequency at baseline and after the addition of carbachol. (C) Quantification of the change in sIPSC frequency with the addition of carbachol. Error bars show mean + SEM. See Supplementary file 1 for Ns. *Significance: *=p < 0.05, **=p < 0.01, ***=p < 0.001, NS = p ≥ 0.05. Abbreviations: sIPSC, spontaneous inhibitory postsynaptic current; CCh, carbachol*.

Figure 6—source data 1 Raw data for quantification in Figure 6B-C and Figure 6—figure supplement 1A, B.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig6-data1-v1.csv
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Figure 6—figure supplement 1

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Additional quantification of sIPSC recordings.

(A) Quantification of average sIPSC amplitude at baseline and after the addition of carbachol. Error bars show mean + SEM. (B) Quantification of the proportion of carbachol-sensitive cells within each genotype. Non-responsive cells are shown in lighter color, responsive cells are shown in darker color. See Supplementary file 1 for Ns. *Significance: *=p < 0.05, **=p < 0.01, ***=p < 0.001, NS = p ≥ 0.05. Abbreviations: sIPSC, spontaneous inhibitory postsynaptic current; CCh, carbachol*.

Figure 7 with 1 supplement

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Reduced seizure induction threshold in models of dystroglycanopathy.

(A) Quantification of latency (in seconds) to generalized tonic clonic seizure upon exposure to 10% flurothyl delivered at a constant rate. Open points denote statistical outliers. See Supplementary file 1 for Ns. *Significance: *=p < 0.05, **=p < 0.01, ***=p < 0.001, NS = p ≥ 0.05*.

Figure 7—source data 1 Raw data for quantification in Figure 7A and Figure 7—figure supplement 1A–C.: https://cdn.elifesciences.org/articles/87965/elife-87965-fig7-data1-v1.csv
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Figure 7—figure supplement 1

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Extended seizure induction threshold data.

(A) Proportion of mice that died as a direct result of a flurothyl-induced seizure. Survivors are depicted in darker colors, fatalities in lighter colors. (B) To minimize the number of animals needed for this experiment, certain genotypes were pooled together. Here we show that pooled genotypes are not different. Statistical tests were run where Ns were sufficient to provide meaningful comparison. (C) Seizure latencies split by sex. Note: only the *Emx1^Cre:Dag1* group is powered sufficiently to statistically compare sexes. Open points denote statistical outliers. Error bars show mean + SEM.

Tables

Table 1

Mouse strains.

Common name	Strain name	Reference	Stock #
Dag1^Flox	B6.129(Cg)-Dag1^tm2.1Kcam/J	Cohn et al., 2002	009652
Dag1^cyto	N/A	Satz et al., 2009	N/A
Pomt2^Flox	POMT2tm1.1Hhu/J	Hu et al., 2011	017880
B4gat1^M155T	B6(C3)-B4GAT1^m1Ddg/J	Wright et al., 2012	022018
Fkrp^P448L	C57BL/6NJ-Fkrp^em1Lgmd/J	Chan et al., 2010	034659
R26^{LSL-H2B-mCherry}	B6.Gt(ROSA)26Sor^tm1.1Ksvo	Peron et al., 2015	023139
Emx1^Cre	B6.129S2-Emx1^tm1(cre)Krj/J	Gorski et al., 2002	005628
Nestin^Cre	B6.Cg-Tg(Nes-cre)^1Kln/J	Tronche et al., 1999	003771
NEX^Cre	NeuroD6^tm1(cre)Kan	Goebbels et al., 2006	MGI:4429523
Sox2^Cre	B6N.Cg-Edil^{3Tg(Sox2-cre)1Amc}/J	Hayashi et al., 2002	014094

Table 2

Breeding schemes.

Breeding Scheme	Control Genotype	Mutant Genotype
Emx1^Cre/+;Dag1^+/- x Dag1^Flox/Flox	Emx1^Cre/+;Dag1^Flox/+	Emx1^Cre/+;Dag1^Flox/-
Nestin^Cre/+;Dag1^+/- x Dag1^Flox/Flox	Nestin^Cre/+;Dag1^Flox/+	Nestin^Cre/+;Dag1^Flox/-
NEX^Cre/+;Dag1^+/- x Dag1^Flox/Flox	NEX^Cre/+;Dag1^Flox/+	NEX^Cre/+;Dag1^Flox/-
Emx1^Cre/+;Pomt2^Flox/+ x Pomt2^Flox/Flox	Emx1^Cre/+;Pomt2^Flox/+	Emx1^Cre/+;Pomt2^Flox/Flox
Dag1^cyto/+ x Dag1^+/-	WT	Dag1^cyto/-
B4gat1^M155T/+ x B4gat1^M155T/+	WT	B4gat1^M155T/M155T
Fkrp^P448L/+ x Fkrp^P448L/+	WT	Fkrp^P448L/P448L

Table 3

Primary antibodies used for immunohistochemistry.

Target	Host species	Dilution	Source	Catalog #	RRID
α-Dystroglycan (IIH6C4)	Mouse	1:250	Millipore	05–593	AB_309828
β-Dystroglycan	Mouse	1:50	Leica Biosystems	NCL-b-DG	AB_442043
CB1R	Guinea pig	1:1000	Synaptic Systems	258–104	AB_2661870
Cux1	Rabbit	1:500	Santa Cruz Biotech	sc-13024	AB_2261231
Laminin	Rabbit	1:1000	Sigma-Aldrich	L9393	AB_477163
NECAB1	Rabbit	1:500	Sigma-Aldrich	HPA023629	AB1848014
NECAB2	Rabbit	1:500	Proteintech	12257–1-AP	AB_2877841
NeuN	Mouse	1:250	Millipore	MAB377	AB_2298772
Parvalbumin	Rabbit	1:1000	Swant	PV27	AB_2631173
Parvalbumin	Mouse	1:50	Swant	235	AB_10000343
Somatostatin	Rabbit	1:2000	Peninsula Labs	T-4103	AB_518614
Tbr1	Rabbit	1:500	Millipore	AB10554	AB_10806888
VGAT	Rabbit	1:500	Synaptic Systems	131–003	AB_887869
VGAT	Guinea Pig	1:500	Synaptic Systems	131–005	AB_1106810
VGlut3	Rabbit	1:1000	Synaptic Systems	135–203	AB_887886
VIP	Rabbit	1:1000	ImmunoStar	20077	AB_572270

Additional files

Supplementary file 1 Experimental N, technical replicates, and sex for experiments.: https://cdn.elifesciences.org/articles/87965/elife-87965-supp1-v1.xlsx
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Supplementary file 2 Statistical tests and analysis for all data.: https://cdn.elifesciences.org/articles/87965/elife-87965-supp2-v1.xlsx
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MDAR checklist: https://cdn.elifesciences.org/articles/87965/elife-87965-mdarchecklist1-v1.pdf
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Jennifer N Jahncke
Daniel S Miller
Milana Krush
Eric Schnell
Kevin M Wright

(2024)

Inhibitory CCK+ basket synapse defects in mouse models of dystroglycanopathy

eLife 12:RP87965.

https://doi.org/10.7554/eLife.87965.3

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Dystroglycan synaptic localization and glycosylation in mouse models of dystroglycanopathy.

Figure 1—source data 1

Emx1Cre drives recombination in forebrain excitatory neurons and astrocytes, but not interneurons or microglia.

Dystroglycan glycosylation is required for synaptic localization.

Figure 1—figure supplement 2—source data 1

Dystroglycan is required for cortical neuron migration in a glycosylation-dependent manner and independent of intracellular interactions.

Figure 2—source data 1

Figure 2—source data 2

Cortical migration is disrupted at midline of B4gat1M155T/M155T but not FkrpP448L/P448L mutants.

Laminin immunoreactivity in adult neocortex appears discontinuous in Dag1 mutants.

Dag1 is required for CCK+/CB1R+ basket IN perisomatic axon targeting in stratum pyramidale of hippocampal CA1-3.

Figure 3—source data 1

CCK+/CB1R+ IN axon targeting phenotypes in hippocampal CA1 of various Dag1 cKOs.

CCK+/CB1R+ IN cell numbers are unchanged in Emx1Cre:Dag1 cKOs.

Figure 3—figure supplement 2—source data 1

Parvalbumin+ basket INs do not require Dag1 for proper axon targeting in hippocampal CA1.

Figure 3—figure supplement 3—source data 1

Altered CB1R expression in cortex and basolateral amygdala of Dag1 and POMT2 mutants.

Dag1 is required for CCK+/CB1R+ IN axon targeting during early postnatal development.

Figure 4—source data 1

Dag1 and Pomt2 cKOs exhibit impaired CB1R+ basket synapse formation in stratum pyramidale of hippocampal CA1.

Figure 5—source data 1

Figure 5—source data 2

Extended quantification of images in Figure 5A–C.

Dag1 and Pomt2 cKOs exhibit increased PV+ basket synapse formation in stratum pyramidale of hippocampal CA1.

Figure 5—figure supplement 2—source data 1

Figure 5—figure supplement 2—source data 2

Extended quantification of images in Figure 5—figure supplement 2A–C.

Dag1 is required for CCK+/CB1R+ IN synapse function in hippocampal CA1 in a manner dependent on both glycosylation and intracellular interactions.

Figure 6—source data 1

Additional quantification of sIPSC recordings.

Reduced seizure induction threshold in models of dystroglycanopathy.

Figure 7—source data 1

Extended seizure induction threshold data.

Mouse strains.

Breeding schemes.

Primary antibodies used for immunohistochemistry.

Supplementary file 1

Supplementary file 2

MDAR checklist

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

Emx1^Cre drives recombination in forebrain excitatory neurons and astrocytes, but not interneurons or microglia.

Cortical migration is disrupted at midline of B4gat1^M155T/M155T but not Fkrp^P448L/P448L mutants.

Dag1 is required for CCK⁺/CB₁R⁺ basket IN perisomatic axon targeting in stratum pyramidale of hippocampal CA1-3.

CCK⁺/CB₁R⁺ IN axon targeting phenotypes in hippocampal CA1 of various Dag1 cKOs.

CCK⁺/CB₁R⁺ IN cell numbers are unchanged in Emx1^Cre:Dag1 cKOs.

Parvalbumin⁺ basket INs do not require Dag1 for proper axon targeting in hippocampal CA1.

Altered CB₁R expression in cortex and basolateral amygdala of Dag1 and POMT2 mutants.

Dag1 is required for CCK⁺/CB₁R⁺ IN axon targeting during early postnatal development.

Dag1 and Pomt2 cKOs exhibit impaired CB1R⁺ basket synapse formation in stratum pyramidale of hippocampal CA1.

Dag1 and Pomt2 cKOs exhibit increased PV⁺ basket synapse formation in stratum pyramidale of hippocampal CA1.

Dag1 is required for CCK⁺/CB₁R⁺ IN synapse function in hippocampal CA1 in a manner dependent on both glycosylation and intracellular interactions.