MeCP2 in cholinergic interneurons of nucleus accumbens regulates fear learning
- Center for Neuroscience and Department of Neurology of Second Affiliated Hospital, Zhejiang University School of Medicine, China
- Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, China
- NHC and CAMS Key Laboratory of Medical Neurobiology, Center for Brain Science and Brain-Inspired Intelligence, Guangdong-Hong Kong-Macao Greater Bay Area, Joint Institute for Genetics and Genome Medicine between Zhejiang University and University of Toronto, Canada
Figures
Figure 1 with 1 supplement
Depletion of MeCP2 in Cholinergic Neurons Resulted in Impaired Fear Regulation.
(A, B) Fluorescence images showing NAc (A) and MS sections (B) in ChatCre mice stained for ChAT, MeCP2, and DAPI. (C, D) Fluorescence images showing NAc (C) and MS sections (D) in Chat-Mecp2-/y mice …
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Figure 1—source data 1
Statistical reporting of Figure 1.
- https://cdn.elifesciences.org/articles/55342/elife-55342-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Chat-Mecp2-/y Mice Showed Normal Ability in Pain Perception, Movement Ability, and Working Memory, Related to Figure 1.
(A) Chat-Mecp2-/y mice showed decreased freezing time compared to combination of other three control groups on Trial four during fear learning. P-values were calculated by two-tailed unpaired t-test.…
Figure 2 with 2 supplements
Restoration of MeCP2 in NAc rather than Cholinergic Neurons of Medial Septum Rescued Fear Deficits in Chat-Mecp2-/y Mice.
(A) Representative fluorescence images showing NAc sections of Chat-Mecp2-/y-AAV/GFP and Chat-Mecp2-/y-AAV/Mecp2 stained for DAPI, MeCP2, and ChAT. (B) Percentage of ChAT-positive cells in NAc …
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Figure 2—source data 1
Statistical reporting of Figure 2.
- https://cdn.elifesciences.org/articles/55342/elife-55342-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Systemic Delivery of Various AAV Vectors into NAc, Related to Figures 2, 4, 5.
(A) Diagram of virus injection into NAc (right) of ChatCre mice and confocal image of AAV/GFP vector-injected NAc slice demonstrating colocalization of GFP expression with ChAT antibody, co-stained …
Figure 2—figure supplement 2
Restoration of MeCP2 in NAc rather than Cholinergic neurons of MS Rescued Fear deficit at the last trial of learning, Related to Figure 2.
(A, B) Fear conditioning test was conducted three weeks after microinjection in NAc (A) or MS (B). P-values were calculated by one-way analysis of variance (ANOVA) with Bonferroni’s multiple …
Figure 3 with 2 supplements
Spontaneous Firing of Cholinergic Interneurons Decreased via Elevated Expression of GABAA α2 Receptors in Chat-Mecp2-/y Mice.
(A, B) Example traces of spontaneous spiking (A) or averaged frequency (B) measured in a cell-attached configuration from NAc cholinergic interneurons. P-values were calculated by one-way analysis …
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Figure 3—source data 1
Statistical reporting of Figure 3.
- https://cdn.elifesciences.org/articles/55342/elife-55342-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Chat-Mecp2-/y Mice Exhibited No Statistical Difference in Intrinsic Properties in NAc Cholinergic Interneurons, Related to Figure 3.
(A–D) Quantitative analysis of resting membrane potential (RMP), membrane time constant (Tau), input resistance (Rin), and membrane capacitance (Cm) of cholinergic interneurons from NAc slices. P-val…
Figure 3—figure supplement 2
Depletion of MeCP2 Did Not Affect mIPSC Frequency of Cholinergic Interneurons, with Limited Expression of GABAA Receptor α1 Subunits in Cholinergic Interneurons within NAc, Related to Figure 3.
(A) Cumulative distribution of mIPSC inter-event interval and average frequency of NAc cholinergic interneurons. P-values were calculated by one-way analysis of variance (ANOVA) with Bonferroni’s …
Figure 4 with 1 supplement
Activation of NAc Cholinergic Interneurons Rescued Fear Deficits in Chat-Mecp2-/y Mice.
(A) Up: Schematic of AAV/hM3Dq viral construct. Down: Confocal image of injected NAc slice demonstrating colocalization of mCherry expression with ChAT antibody, co-stained with DAPI. (B, C) Fear …
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Figure 4—source data 1
Statistical reporting of Figure 4.
- https://cdn.elifesciences.org/articles/55342/elife-55342-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Activation of NAc Cholinergic Interneurons Rescued Fear Deficits in Chat-Mecp2-/y Mice, Related to Figure 4.
(A, B) Representative spontaneous firing (A) and summary histogram (B) of hM3Dq-mCherry expressing ChAT neurons in Chat-Mecp2-/y mice before (red) and after (gray) CNO treatment. P-values were …
Figure 5 with 1 supplement
Inhibition of NAc Cholinergic Interneurons in ChatCre Mice Mimicked Fear Deficits in Chat-Mecp2-/y Mice.
(A) Schematic of AAV/hM4Di viral construct. Confocal image of injected NAc slice demonstrating colocalization of mCherry expression with ChAT antibody, co-stained with DAPI. (B, C) Fear retrieval …
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Figure 5—source data 1
Statistical reporting of Figure 5.
- https://cdn.elifesciences.org/articles/55342/elife-55342-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Inhibition of NAc Cholinergic Interneurons in ChatCre Mice, Related to Figure 5.
Confocal image of injected NAc slice demonstrating localization of NpHR expression and fiber position in NAc core (A), NAc ventral medial shell (B), and NAc medial shell (C).
Figure 6 with 2 supplements
Manipulation of α2-GABAA Receptors in NAc Mimicked Fear Deficits in Chat-Mecp2-/y Mice.
(A) Schematic of AAV/RNAi viral construct. Confocal image of injected NAc slice demonstrating colocalization of GFP expression with ChAT antibody, co-stained with DAPI. (B, C) mIPSCs of …
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Figure 6—source data 1
Statistical reporting of Figure 6.
- https://cdn.elifesciences.org/articles/55342/elife-55342-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Knockdown of α2-GABAA Receptors in Cholinergic Interneurons Did Not Affect mIPSC Frequency in Cholinergic Interneurons, Related to Figure 6.
Cumulative distribution of mIPSC amplitude in NAc neurons infected by AAV-Sham or AAV/RNAi. P-values were calculated by one-way analysis of variance (ANOVA) with Bonferroni’s multiple comparisons …
Figure 6—figure supplement 2
Retrograde tracing from NAc cholinergic neurons.
(A) Sequential injections of AAV-FLEX-TVA-GFP, AAV-FLEX-Rabies-glycoprotein and G-deleted EnA-rabies-mCh into NAc of a ChatCre mouse labeled starter cells (yellow). (B, C) Transsynaptic retrograde …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (M. musculus) | ChatCre | Jackson Laboratory | Jax No.006410 | |
| Genetic reagent (M. musculus) | MeCP2 Flox/- | Jackson Laboratory | Jax No.006847 | |
| Antibody | Goat polyclonal anti-ChAT | Millipore | Cat# AB144P | IHC: 1:100 |
| Antibody | Rabbit monoclonal anti-MeCP2 | Cell Signaling Technology | Cat#3456 s | IHC: 1:200 |
| Antibody | Mouse monoclonal anti-PV | Swant | Cat#PV235 | IHC: 1:1 000 |
| Antibody | Rabbit polyclonal GABAA receptor α1 | Millipore | Cat#06–868 | IHC: 1:1 000 WB: 1:500 |
| Antibody | Rabbit polyclonal GABAA receptor α2 | Synaptic System | Cat#224 103 | IHC: 1:1 000 |
| Antibody | Rabbit polyclonal GABAA receptor α2 | Abcam | Cat#ab72445 | WB: 1:500 |
| Antibody | Rabbit monoclonal anti-GAPDH | Cell Signaling Technology | Cat#5014S | WB: 1:5000 |
