Leptin suppresses development of GLP-1 inputs to the paraventricular nucleus of the hypothalamus
- Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, United States
- Department of Pharmacology, University of Virginia School of Medicine, United States
Figures
Figure 1
Ontogeny of GLP-1 projections into the PVH.
Maximum intensity projections of confocal images through the PVH of WT (A–E) and Lepob/ob (F–J) mice illustrate the density of GLP-1 immunoreactive fibers (green) present at postnatal ages P6, P10, …
Figure 2 with 1 supplement
Leptin receptor-expressing neurons in the NTS are responsive to leptin during early postnatal development and project to the PVH.
(A), (B) Representative images of pSTAT3 immunoreactivity (green) in LepRb-Cre::tdTom mice injected with leptin or saline control at P16. (C), (D) Representative images of pSTAT3 immunoreactivity in …
Figure 2—figure supplement 1
Distribution of GCG-Cre::SynTom expression in the hindbrain and hypothalamus.
GCG-Cre mice were crossed with mice carrying a fluorescent reporter synaptophysin-tdTomato fusion protein to visualize PPG neurons and their inputs. Neurons expressing GCG-Cre::SynTom (PPG neurons) …
Figure 3
Axonal labeling of projections from PPG neurons to the PVH are increased in leptin-deficient mice.
Genetic targeting of SynTom fluorescence to axons derived from PPG neurons revealed a significant increase in the density of GLP-1 projections to the PVHmpd and PVHpml in Lepob/ob mice. Quantitative …
Figure 4
Target-specific enhancement of GLP-1 inputs to CRH neurons in leptin-deficient mice.
Images (maximum intensity projections of confocal images) showing GLP-1 axonal labeling (red) onto Oxytocin- (A,B), or CRH-immunoreactive (D,E) neurons (green) in the PVH of GCG-Cre::SynTom::WT and …
Figure 5
Viscerosensory transmission to the PVH is disrupted in leptin-deficient mice.
Compared with saline-injected controls (A), i.p. injection of CCK increased the number of Fos immunoreactive nuclei (green) in the PVH (B,C), and within PPG-labeled neurons (red) in the NTS (insets) …
Figure 6
Cell-autonomous action of leptin in the NTS specifies the density and activity of GLP-1 inputs to PVH neurons.
Compared with WT mice (A), mice functionally null for LepRb (LepRbTB/TB mice) exhibited increased GLP-1-immunoreactive fiber density in both the mpd and pml compartments of the PVH (B), similar to …
Figure 7
Leptin’s action on PPG neurons is not sufficient to restore food intake and anxiety-like behavior in leptin receptor-deficient mice.
Compared with WT controls, LepRbTB/TB mice displayed increased body weight (A), increased total pellets consumed (B), decreased meal number (C) and increased meal size (D). Restoration of LepRb …
Figure 8
Leptin is required to specify the normal balance of hormonal and viscerosensory information conveyed by AgRP and GLP-1 inputs that converge onto the PVH.
During development, the adipocyte-derived hormone leptin is secreted by adipose tissue and binds to leptin receptors (LepRb) located on AgRP neurons in the ARH, and on PPG neurons in the NTS. In …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (M. musculus) | tdTomato | Jackson Laboratory | Stock #:007914 RRID:IMSR_JAX:007914 | PMID:20023653 |
| Genetic reagent (M. musculus) | Gcg-Cre | PMID:28218622 | RRID:MMRRC_051056 | Dr. Michael Scott (University of Virginia) |
| Genetic reagent (M. musculus) | LepRb-Cre | Jackson Laboratory | Stock #:032457 RRID:IMSR_JAX:032457 | Dr. Martin Myers, Jr. (University of Michigan) PMID:17021368 |
| Genetic reagent (M. musculus) | Lepob/ob | Jackson Laboratory | Stock #:000632 RRID:IMSR_JAX:000632 | Lepob/+ heterozygotes were bred to generate homozygous, leptin-deficient mice |
| Genetic reagent (M. musculus) | LepRTB/TB | Jackson Laboratory | Stock #:018989 RRID:IMSR_JAX:018989 | PMID:22326958 |
| Genetic reagent (M. musculus) | Glp1r-Cre | Jackson Laboratory | Stock #:029283 RRID:IMSR_JAX:029283 | PMID:27238020 |
| Genetic reagent (M. musculus) | Synaptophysin-tdTomato | Jackson Laboratory | Stock #:012570 RRID:IMSR_JAX:012570 | MGI ID: J:170755 |
| Antibody | Rabbit monoclonal anti-cFos | Cell Signaling | Cat. #: 2250S RRID:AB_2247211 | IHC (1:1000) |
| Antibody | Rabbit polyclonal anti-CRF | Dr. Paul Sawchenko; Dr. Wylie Vale (Salk Institute) | PBL#rC68 | IHC (1:1000) |
| Antibody | Mouse monoclonal anti-HuC/D | Molecular Probes | Cat. #: A21271 RRID:AB_221448 | IHC (1:500) |
| Antibody | Rabbit polyclonal anti-dsRed | Clontech | Cat. #: 632496 RRID:AB_10013483 | IHC (1:1000) |
| Antibody | Rabbit anti-Oxytocin | Peninsula | Cat. #: T-4084 RRID:AB_518524 | IHC (1:2500) |
| Antibody | Rabbit polyclonal anti-GLP-1 | Peninsula | Cat. #: T-4363 RRID:AB_518978 | IHC (1:5000) |
| Antibody | Rabbit polyclonal anti-pSTAT3 (Y705) | Cell Signaling | Cat. #: 9131S RRID:AB_331586 | IHC (1:1000) |
| Antibody | Goat polyclonal anti-rabbit Alexa Fluor 568 | ThermoFisher Scientific | Cat. #: A-11036 RRID:AB_10563566 | IHC (1:500) |
| Antibody | Goat polyclonal anti-rabbit Alexa Fluor 488 | ThermoFisher Scientific | Cat. #: A-11034 RRID:AB_2576217 | IHC (1:500) |
| Antibody | Goat polyclonal anti-mouse Alexa Fluor 647 | ThermoFisher Scientific | Cat. #: A-21236 RRID:AB_2535805 | IHC (1:500) |
| Recombinant DNA reagent | AAV pCAG-FLEX-EGFP-WPRE | Addgene | Cat. #: 51502 RRID:Addgene_51502 | PMID:24695228 |
| Peptide, recombinant protein | Leptin | Peprotech | Cat. #: 450–31 | 10 mg/kg i.p. |
| Peptide, recombinant protein | CCK | Bachem | Cat. #: H-2080 | 10 μg/kg i.p. |
| Commercial assay or kit | Corticosterone RIA | MP Biomedicals | Cat. #: 07–120102 | |
| Chemical compound, drug | Colchicine | Sigma | Cat. #: C9754 | 4 mg/mL |
| Software, algorithm | Imaris | Bitplane | V9.2 | |
| Software, algorithm | Volocity | PerkinElmer | V6.1.1 | |
| Software, algorithm | GraphPad Prism | Prism | Prism 7 |
