A limbic circuit selectively links active escape to food suppression
- Laboratory of Molecular Genetics, The Rockefeller University, United States
- Department of Psychiatry, Weill Cornell Medical College-New York Presbyterian Hospital, United States
- Howard Hughes Medical Institute, The Rockefeller University, United States
Figures
Figure 1 with 1 supplement
Identification of lateral septum neurons activated by acute restraint stress.
(A) Activity mapping of the whole brain using c-fos staining in control (naïve) and restraint-stressed (RS) mice. Areas with the highest c-fos expression (green) difference are shown: the basolateral…
Figure 1—figure supplement 1
LSNTS neurons are activated by acute restraint stress.
(A) Representative images of c-fos (green) in the lateral septum of control and restraint stress (RS) mice. Right, quantification of c-fos+ cells in control (black) and RS (red) mice. n = 4, …
Figure 2 with 2 supplements
LSNts neurons regulate food intake and body weight in mice.
(A) Left panel, experimental scheme. Right panel, food intake, measured in grams (g), in control (black) and RS (red) mice after 60 and 120 min following RS. n = 5(RS), n = 6(control), Two-way ANOVA …
Figure 2—figure supplement 1
LSNts neurons regulate feeding.
(A) Representative images of viral injection strategy and viral expression (mCherry, red) and c-fos (green) in control virus (left) and hM3Dq (right) injected mice following CNO injection. Right, …
Figure 2—figure supplement 2
LSNts neurons do not regulate anxiety-like behaviors.
(A) Time, measured in seconds (s), spent in the center and border of an open field in control mCherry (black) and hM3Dq (red) expressing Nts-cre mice. n = 8(mCherry), n = 9(hM3Dq). Two-way ANOVA …
Figure 3 with 1 supplement
Molecular and functional profiling of LSNts neurons.
(A) Experimental scheme of the viralTrap experiment. Nts-cre mice are injected with an AAV expressing cre-dependent GFPL10 (AAV-Introvert--GFPL10) into the LS and then GFP+ polysomes are …
Figure 3—figure supplement 1
Regulation of feeding and/or anxiety by Cartpt-, Glp1r- and Sst-expressing neurons in the lateral septum.
(A) Experimental scheme. Wild-type mice are injected with Ex4 or Mc3r agonist y-MSH directly into the LS or Glp1r-cre, Cartpt-Cre, or Sst-cre mice with viral expression of hM3Dq activatory DREADD …
Figure 4 with 1 supplement
LSNts- > LH projection regulates food intake in mice.
(A) Representative images of the LS injection site (left) and terminals in the lateral hypothalamus (left) of Nts-cre mice injected with a control mCherry reporter virus. Atlas images above the …
Figure 4—figure supplement 1
LSNts → LH does not regulate anxiety or locomotion.
(A) Left, experimental scheme of Glp1r-cre mice injected with AAV expressing cre-dependent mCherry in the lateral septum. Right, representative image of Glp1r fibers in the lateral hypothalamus. …
Figure 5 with 1 supplement
LSNts neurons are specifically activated by stressful situations involving active coping.
(A) Left panel, experimental scheme: mice were manually immobilized for the period of 1 min. Ca+2 activity was recorded before, during, and after the period of manual immobilization. Middle, Average …
Figure 5—figure supplement 1
LSNts neurons are not selectively tuned to feeding or movement.
(A) Experimental scheme. Mice were then attached to a fiber photometry setup with a 465 nm LED (blue) and a 405 nm LED (purple) and a photodetector. (B) Representative image of viral expression in …
Videos
Video 1
Representative video of the simulated ‘predator’ test.
The mouse is placed in the open field with a designated safe zone. A robotic spider ‘predator’ is introduced and ‘attacks’ as the mouse exits the safe zone. Calcium recordings were taken throughout …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Mus musculus) | Mouse: Nts-cre | Jackson Laboratories | Stock#017525 | N/A |
| Genetic reagent (Mus musculus) | Mouse: Cartpt-cre | Jackson Laboratories | Stock#028533 | N/A |
| Genetic reagent (Mus musculus) | Mouse: Sst-cre | Jackson Laboratories | Stock#013044 | N/A |
| Genetic reagent (Mus musculus) | Mouse: Glp1r-cre | Jackson Laboratories | Stock#029283 | N/A |
| Antibody | Anti-phospho ribosomal protein 6 pSer244/pSer247 (Rabbit polyclonal) | Invitrogen | Cat#44–923G, RRID:AB_2533798 | (1:1000) |
| Antibody | Anti-GFP (Chicken polyclonal) | Abcam | Cat#ab13970, RRID:AB_300798 | (1:1000) |
| Antibody | 19C8 and 19F7, GFP monoclonal antibodies | Memorial Sloan-Kettering Monoclonal Antibody Facility | Custom order | (50 ug) |
| Antibody | Anti-mCherry (Chicken polyclonal) | Abcam | Cat# ab205402 | (1:1000) |
| Antibody | Mouse anti-cfos (Rabbit monoclonal) | Cell Signaling | Cat#2250S, RRID:AB_2247211 | (1:500) |
| Antibody | Anti-rabbit IgG Alexa 488 (Goat polyclonal) | Invitrogen | Cat#A11008, RRID:AB_143165 | (1:1000) |
| Antibody | Anti-rabbit IgG Alexa 594 (Goat polyclonal) | Invitrogen | Cat#A11072, RRID:AB_142057 | (1:1000) |
| Antibody | Anti-chicken IgG Alexa 488 (Goat polyclonal) | Invitrogen | Cat#A11039, RRID:AB_142924 | (1:1000) |
| Antibody | Anti-chicken IgG Alexa 594 (Goat polyclonal) | Invitrogen | Cat#A11042, RRID:AB_142083 | (1:1000) |
| Recombinant DNA reagent | AAV5-EF1a-DIO-YFP | UNC Vector Core | N/A | N/A |
| recombinant DNA reagent | AAV5-EF1a-DIO-hChR2(H134R)-YFP | UNC Vector Core | N/A | N/A |
| Recombinant DNA reagent | AAV5-EF1a-DIO-hM3D(Gq)-mCherry | UNC Vector Core | N/A | N/A |
| Recombinant DNA reagent | AAV5-EF1a-DIO-mCherry | UNC Vector Core | N/A | N/A |
| Recombinant DNA reagent | AAV5-EF1a- DIO-hM4D(Gi)-mCherry | UNC Vector Core | N/A | N/A |
| Recombinant DNA reagent | AAV5-DIO-GCaMP6s | Addgene | Cat#510882 | N/A |
| Recombinant DNA reagent | HSV-lsl-tdTomato | Lo and Anderson, 2011 | N/A | N/A |
| Recombinant DNA reagent | AAV5-Introvert-GFPL10a | Nectow et al., 2017 | N/A | N/A |
| Sequence-based reagent | PrimeTime Standard qPCR Assay for NTS | idtDNA | Mm.PT.58.10351472 | N/A |
| Sequence-based reagent | PrimeTime Standard qPCR Assay for ActB | idtDNA | Mm.PT.47.5885043.g | N/A |
| Sequence-based reagent | Probe for NTS (C1) | ACDBio | Cat# 420448 | N/A |
| Sequence-based reagent | Probe for cFos (C2) | ACDBio | Cat#506921-C2 | N/A |
| Sequence-based reagent | Probe for Glp1r (C3) | ACDBio | Cat#418851-C3 | N/A |
| Sequence-based reagent | Probe for Sst (C3) | ACDBio | Cat#404631-C3 | N/A |
| Sequence-based reagent | Probe for Mc3r (C3) | ACDBio | Cat#412541-C3 | N/A |
| Sequence-based reagent | Probe for Cartpt (C3) | ACDBio | Cat#432001-C3 | N/A |
| Sequence-based reagent | Probe for vGAT (C2) | ACDBio | Cat#319191-C2 | N/A |
| Sequence-based reagent | Probe for vGLUT2 (C3) | ACDBio | Cat#319171-C3 | N/A |
| Peptide, recombinant protein | NBL10 recombinant peptide | Chromotek | Cat# GT-250 | 100 ng/mL |
| Commercial assay or kit | RNAeasy Mini kit | Qiagen | Cat#74104 | N/A |
| Commercial assay or kit | RNAscope Fluorescent Multiplex Reagent Kit V2 | ACDbio | Cat#323100 | N/A |
| Chemical compound, drug | Clozapine-N-oxide(CNO) | Tocris Biosciences | Cat#4936; CAS: 34233-69-7 | 1 mg/kg |
| Software, algorithm | Tophat | Basespace | https://basespace.illumina.com/apps/ | N/A |
| Software, algorithm | Cufflinks | Basespace | https://basespace.illumina.com/apps/ | N/A |
| Software, algorithm | SYNAPSE | Tucker-Davis Technologies | https://tdt.com | N/A |
| Software, algorithm | MATLAB | Mathworks | https://mathworks.com | N/A |
| Software, algorithm | ImageJ | NIH | https://imagej.nih.gov/ij/ | N/A |
| Software, algorithm | GraphPad Prism 5.0 | GraphPad | https://www.graphpad.com/scientific-software/prism/ | N/A |
| Software, algorithm | Ethovision 9.0 | Noldus | https://noldus.com | N/A |
Additional files
-
Supplementary file 1
c-Fos expression in the mouse brain after restraint stress.
Expression is noted as − (no expression), + (low expression), ++ (moderate expression) and +++ (high expression).
- https://cdn.elifesciences.org/articles/58894/elife-58894-supp1-v1.docx
-
Supplementary file 2
Phospho-TRAP differential gene expression data.
Data is sorted by fold-change (log2) in restraint stress (RS) samples compared to Naïve (N) samples along with the corresponding q-values.
- https://cdn.elifesciences.org/articles/58894/elife-58894-supp2-v1.xlsx
-
Supplementary file 3
Viral-TRAP differential gene expression data.
Significantly enriched (TRUE) genes are shown and are sorted by fold-change (log2).
- https://cdn.elifesciences.org/articles/58894/elife-58894-supp3-v1.xlsx
-
Transparent reporting form
- https://cdn.elifesciences.org/articles/58894/elife-58894-transrepform-v1.pdf
