NPAS4 recruits CCK basket cell synapses and enhances cannabinoid-sensitive inhibition in the mouse hippocampus
- University of California San Diego, United States
Figures
Figure 1 with 1 supplement
NPAS4 enhances somatic inhibition onto superficial CA1 pyramidal neurons.
(A) Cartoon of standard environment (SE) and enriched environment (EE). (B) Confocal images of mouse hippocampi from SE (top) and EE (bottom) stained with antibodies recognizing NPAS4 (cyan) and …
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Figure 1—source data 1
NPAS4 enhances somatic inhibition onto superficial CA1 PNs.
- https://doi.org/10.7554/eLife.35927.005
Figure 1—figure supplement 1
Superficial and deep CA1 PNs express NPAS4 equivalently after exploration of an EE.
Quantification of NPAS4-positive neurons in superficial and deep CA1 PNs in mice from SE and EE (n = 10 sections from a total of 3 mice per condition). * indicates p<0.05.
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Figure 1—figure supplement 1—source data 1
NPAS4 expression in superficial and deep CA1.
- https://doi.org/10.7554/eLife.35927.004
Figure 2 with 1 supplement
NPAS4 regulates CCKBC input onto PNs in superficial CA1.
(A) Schematic representation of PN with CCKBC and PVBC synaptic input. CCKBC boutons contain CB1Rs (blue) and utilize N-type VGCCs for neurotransmission. (B) Representative images of tissues stained …
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Figure 2—source data 1
NPAS4 regulates CCKBC input onto PNs in superficial CA1.
- https://doi.org/10.7554/eLife.35927.009
Figure 2—figure supplement 1
Experience-induced Npas4 does not affect CCKBC input onto deep CA1 PNs.
ω-AgTx-IVA (0.3 μM) is used to isolate synaptic release from CCKBCs. Example eIPSCs from WT (black) and Npas4 KO (green) PNs in deep CA1 (left) of mice housed in EE. Pairwise comparison of eIPSCs …
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Figure 2—figure supplement 1—source data 1
Experience-induced Npas4 does not affect CCKBC input onto deep CA1 PNs.
- https://doi.org/10.7554/eLife.35927.008
Figure 3 with 1 supplement
NPAS4 does not regulate PVBC input onto PNs in CA1.
(A) Schematic representation of PN with CCKBC and PVBC synaptic input. PVBCs express PV (blue) and utilize P/Q-type VGCCs for neurotransmission. (B) Representative images of tissues stained with …
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Figure 3—source data 1
NPAS4 does not regulate PVBC input onto PNs in CA1.
- https://doi.org/10.7554/eLife.35927.013
Figure 3—figure supplement 1
Experience-induced NPAS4 does not affect PVBC input onto deep CA1 PNs.
ω-Ctx-GVIA (1 μM) is used to isolate synaptic release from PVBCs. Example eIPSCs from WT (black) and Npas4 KO (green) PNs in deep CA1 (left) of mice housed in EE. Pairwise comparison of eIPSCs …
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Figure 3—figure supplement 1—source data 1
Experience-induced NPAS4 does not affect PVBC input onto deep CA1 PNs.
- https://doi.org/10.7554/eLife.35927.012
Figure 4 with 1 supplement
Enriched environment increases inhibition of CA1 PNs by CCKBCs but not PVBCs.
(A–C) Electrophysiological characteristics of PV inhibitory neurons (INs) and CCK INs: (A) example spike trains (scale bar = 50 ms, 10 mV), (B) AP FWHM) n=17 for PV INs, n=20 for CCK INs), *** …
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Figure 4—source data 1
Distinguishing CCKBCs from CCK SCA interneurons.
- https://doi.org/10.7554/eLife.35927.016
Figure 4—figure supplement 1
CCKBCs were differentiated from dendritic Schaffer collateral-associated (SCA) interneurons by synaptic properties and morphology.
(A) Examples of presynaptic APs (top) and postsynaptic uIPSCs (bottom) recorded from somatic (left) and dendritic-targeting (right) INs and PNs, respectively. Scale bar = 25 ms, 50 pA. (B) uIPSC …
Figure 5 with 1 supplement
NPAS4 regulates the number of CCKBC synapses onto PNs but does not alter synaptic properties.
(A) Schematic of recording from synaptically connected CCKBC and WT or Npas4 KO PN pairs. (B) Examples of CCKBC APs (top) and PN uIPSCs (bottom) recorded from WT and Npas4 KO PNs. Scale bars: top = 2…
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Figure 5—source data 1
Properties of CCKBC synapses made onto WT and Npas4 KO PNs.
- https://doi.org/10.7554/eLife.35927.020
Figure 5—figure supplement 1
CCKBC–PN pairs with WT and KO PNs have similar uIPSC onset times.
Latency to uIPSC onset between CCKBC–PN pairs with WT and Npas4 KO PNs (WT: n=12 pairs, KO: n=10 pairs). Example AP from a CCKBC (black) and average uIPSCs from WT (gray) and Npas4 KO (green) PNs. …
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Figure 5—figure supplement 1—source data 1
uIPSC onset times.
- https://doi.org/10.7554/eLife.35927.019
Figure 6
Experience-driven NPAS4 expression enhances the magnitude of DSI in PNs.
(A) Schematic of recording configuration; somatic and dendritic inhibitory inputs are represented. Whole-cell patch clamp recordings were obtained from WT (shown) or Npas4 KO PNs. Spontaneous IPSCs …
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Figure 6—source data 1
DSI in WT and Npas4 KO PNs.
- https://doi.org/10.7554/eLife.35927.022
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Mus musculus) | Npas4 | NA | Gene ID: 225872 | |
| Genetic reagent (Mus musculus) | Npas4f/f | Michael Greenberg (HMS) | ||
| Genetic reagent (Adeno-associated virus) | Cre-GFP | Penn Vector Core | AAV1.hSyn.HI.eGFP- Cre.WPRE.SV40 | |
| Genetic reagent (Adeno-associated virus) | mRFP | Penn Vector Core | AAV1.hSyn.TurboRFP.WPRE.rBG | |
| Antibody | anti-Npas4 | Michael Greenberg (HMS) | 1 to 1000 | |
| Antibody | anti-Gephyrin | Synaptic Systems | RRID:AB_2619834 | 1 to 200 |
| Antibody | anti-VGAT | Synaptic Systems | RRID:AB_887873 | 1 to 350 |
| Antibody | anti-CB1R | Synaptic Systems | RRID: AB_2619970 | 1 to 500 |
| Antibody | anti-parvalbumin | Abcam | RRID:AB_298032 | 1 to 1000 |
| Antibody | anti-NeuN | Synaptic Systems | RRID:AB_2619988 | 1 to 1000 |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.35927.023
