Somatostatin interneurons activated by 5-HT2A receptor suppress slow oscillations in medial entorhinal cortex
- Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health; Neuroscience Research Center, Germany
- Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health; Cluster of Excellence NeuroCure, Germany
- German Centre for Neurodegenerative Diseases (DZNE), Germany
- Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health; Department of Experimental Neurology, Germany
- Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health; Center for Stroke Research Berlin, Germany
- Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health; Einstein Center for Neurosciences Berlin, Germany
Figures
Figure 1 with 3 supplements
MDMA/Fen inhibit SOs in anesthetized mice.
(A) Immunocytochemical analysis of an ePet-YFP mouse showing serotonergic fibers in medial entorhinal cortex, horizontal slice (M = medial, L = lateral, P = posterior, A = anterior). Scale bar: 20 …
Figure 1—figure supplement 1
In vivo upstate spatial features.
(A) Microelectrode implant location and microelectrode features. Sixty-four channels (nanimals = 15, nshanks = 4) and 32 channels (nanimals = 3, nshanks = 2) microelectrodes were used in this study, …
Figure 1—figure supplement 2
LFP power analysis for saline and MDMA/Fen injection.
(A) Average normalized LFP power during baseline and saline injection. (B) Average LFP power during baseline and MDMA/Fen injection. (C) Box plots of normalized LFP power per frequency band: delta …
Figure 1—figure supplement 3
In vivo upstate metrics for saline and MDMA/Fen injection.
(A) Average upstate voltage deflection for control and MDMA/Fen, patches represent one standard deviation. Scale bars: 0.5 s, 200 µV. (B) Violin plots of duration (control: 0.98 ± 0.02 s, MDMA/ Fen: …
Figure 2 with 2 supplements
Divergent unit responses to MDMA/Fen application.
(A) Spike rate of the activated units versus all the other units during MDMA/Fen application (activated: n = 31, Non-activated: n = 324). (B) Top: TP latencies color-coded by group. Middle: …
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Figure 2—source data 1
Source data for Figure 2A.
- https://cdn.elifesciences.org/articles/66960/elife-66960-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Cross-correlogram (CCG) based connectivity analysis.
(A) Units are plotted according to TP latencies and repolarization time and color-coded according to the number of inhibitory connections detected. Units displaying a TP latency <0.55 ms were …
Figure 2—figure supplement 2
TP latency density distributions.
(A) Dashed lines represent kernel density estimations of probability density functions of the TP latencies of putative inhibitory (blue), putative excitatory (black) and ‘activated’ units. Solid …
Figure 3 with 9 supplements
5-HT suppresses SOs and activates Sst interneurons.
(A) Biocytin staining of four simultaneously recorded cells shown in (B) WFS1 expression (in red) delimits L2/3 border. (B) Intracellular recordings showing synchronous upstate events in four …
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Figure 3—source data 1
Source data for Figure 3G.
- https://cdn.elifesciences.org/articles/66960/elife-66960-fig3-data1-v2.xlsx
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Figure 3—source data 2
Source data for Figure 3L.
- https://cdn.elifesciences.org/articles/66960/elife-66960-fig3-data2-v2.xlsx
Figure 3—figure supplement 1
Effect of Fen on SOs in vitro.
(A) Top: upstate raster plot during Fen application. Bottom: Histogram of upstate incidence before and after Fen application (n = 6, baseline: 1.13 ± 0.17 upstates/10 s, Fen: 0.04 ± 0.03 upstates/10 …
Figure 3—figure supplement 2
5-HT suppresses evoked upstates in vitro.
(A) Experimental protocol: recording and stimulation electrode were placed in mEC layer 3, stimulation electrode was located toward the lateral side of the slice. (B) Effect of electrical …
Figure 3—figure supplement 3
5-HT2ARs are involved in 5-HT mediated SOs suppression in vitro.
(A) Top: upstate raster plot during application of WAY 100635 (5-HT1A antagonist) + 5-HT. Bottom: change in RP in putative excitatory cells after application of WAY 100635 (5-HT1A antagonist) + 5-HT …
Figure 3—figure supplement 4
5-HT3 receptor is not involved in 5-HT mediated SOs suppression.
(A) Top: upstate raster plot during application of m-CPBG (5-HT3 agonist). Bottom: histogram of upstate incidence before and after m-CPBG application (n = 6, baseline: 1.04 ± 0.19 upstates/10 s, …
Figure 3—figure supplement 5
In vitro upstates metrics during baseline and ketanserin + 5-HT application.
(A) Box plot showing spiking rate before and after ketanserin + 5-HT (baseline: 0.38 ± 0.14 spikes/s, ketanserin + 5-HT: 0.15 ± 0.06 spikes/s, p=0.011, Mann-Whitney test). patches represent 95% …
Figure 3—figure supplement 6
LTS neurons are depolarized by 5-HT.
(A) PCA projection plot of all the cells recorded. Cells are color-coded according to group identity (posterior probability >0.9): excitatory (black), fast-spiking (blue), low-threshold spiking …
Figure 3—figure supplement 7
Excitatory, fast-spiking, and LTS cells have unique sets of electrophysiological features.
(A) Box plot showing the values of input resistance (Rin), delta after-hyperpolarization (ΔAHP), SAG, RP, rheobase and spike width of excitatory (Exc, black), fast-spiking (FS, blue) and …
Figure 3—figure supplement 8
Classification of cells recorded in Sst-tdTomato mice.
(A) PCA projection plot of cells recorded in Sst-tdTomato mice. Cells are color-coded according to group identity (posterior probability >0.9): fast-spiking (blue) or low-threshold spiking (green). …
Figure 3—figure supplement 9
Spatial localization of 5-HT2AR positive cells in EC.
(A) 3D visualization of EC (purple). (B) 3D localization of all the 5-HT2AR-positive cells detected in EC using same perspective as (A). (Cand D) Co-localization of 5-HT2AR and Sst, arrows point to …
Figure 4 with 1 supplement
Sst interneurons activation suppresses SOs.
(A) Experimental protocol: Sst interneurons expressing ChR2 are activated by light during intracellular recording of L3 neurons in EC. (B) Representative recordings from a L3 neuron during Sst …
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Figure 4—source data 1
Source data for Figure 4E.
- https://cdn.elifesciences.org/articles/66960/elife-66960-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Vector construction and RMCE for the generation of a transgenic mouse line with Cre-conditional hM4Di expression.
(A) The coding sequence of hM4Di-mKate flanked by two opposing loxP and lox2272 sites was placed in reverse orientation to the CAG- promoter in the pRMCE. In the acceptor ES cells the ROSA26 allele …
Figure 5 with 2 supplements
Sst interneurons mediate the effect of 5-HT on SOs.
( A) Upstate raster plot during 5-HT and subsequent CNO application. Orange box represents 5-HT, purple boxes represent CNO. Note the appearance of upstates after CNO application. (B) Upstate …
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Figure 5—source data 1
Source data for Figure 5B.
- https://cdn.elifesciences.org/articles/66960/elife-66960-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
CNO application prevents 5-HT mediated upstates suppression.
(A) Upstate raster plot during CNO and subsequent 5_HT application. Orange box represents 5-HT, purple boxes represent CNO. CNO significantly prevents 5-HT-induced suppression of upstates. (B) …
Figure 5—figure supplement 2
CNO application in wild-type littermates and PV-hM4Di mice.
(A) CNO application in wild-type littermates. Top: Experimental protocol. Orange box represents 5-HT and purple boxes represent CNO application. Bottom: upstate incidence during 5-HT and 5-HT+CNO …
Author response image 1
SO are not suppressed by optogenetic activation of Raphe 5-HT neurons during anesthesia.
(A) DRN immunohistochemistry image showing ChR2-YFP infection in DRN serotonergic neurons. 5-HT in red, YFP in green. Scale bars: 50 µm. (B) 3D visualization of microelectrodes location for all …
