A neurogenic signature involving monoamine Oxidase-A controls human thermogenic adipose tissue development
- Morningside Graduate School of Biomedical Sciences, University of Massachusetts Medical School, United States
- Program in Molecular Medicine, University of Massachusetts Medical School, United States
- Department of Surgery, University of Massachusetts Medical School, United States
- Diabetes Center of Excellence, University of Massachusetts Medical Center, United States
Figures
Figure 1 with 1 supplement
Characteristics of adipocytes during development of WIM and TIM.
(A) Representative images of PLIN1 (green) and DAPI (blue) staining of WIM at the indicated weeks after implantation, and of inguinal mouse and subcutaneous human adipose tissue, scale bar = 50 µm. …
Figure 1—figure supplement 1
Development of implants from cultured adipocytes in mice.
(A) Representative images of differentiated adipocytes and single cell suspension used for implantation, scale bar = 25 µm. (B) Representative image of bilateral flank implantation, scalebar = 0.5 …
Figure 2 with 1 supplement
Vascular development of WIM and TIM.
(A) Representative images of isolectin staining of WIM at the indicated weeks after implantation, and of inguinal mouse and subcutaneous human adipose tissue, scale bar = 50 µm. (B) Total isolectin …
Figure 2—figure supplement 1
Density of macrophages in WIM and TIM.
(A) Representative images of thin sections WIM and TIM after 8 weeks of implantation, stained for F4/80 (red), CD45 (green) and DAPI (blue). (B) Macrophage staining, where symbols are the mean of …
Figure 3
Capacity of thermogenic adipocytes to autonomously maintain their phenotype, and their effect on host adipose depots.
(A) Experimental paradigm for in vitro experiments. (B) Phase images of cultured adipocytes during chronic stimulation and withdrawal. (C) Quantification of droplet size, where symbols are the mean …
Figure 4
Distinct gene expression signatures of adipocytes and stromal cells in WIM and TIM.
(A) Overview of analysis paradigm. (B) Dimensional reduction plot of data from Burl et al., 2018, using Uniform Manifold Approximation and Projection (UMAP), where ASC = adipose stem cells, Diff. …
Figure 5
Differentially expressed genes persisting through tissue development.
(A) Volcano plot of genes differentially expressed between non-thermogenic and thermogenic adipocytes prior to implantation. Highlighted are selected genes characteristic of human thermogenic …
Figure 6 with 1 supplement
Species specific expression of MAOA in adipocytes.
(A) Dot plots of indicated genes generated using data from Emont et al., 2022, available here (B) RT-PCR and western blotting at indicated days following induction of differentiation in human …
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Figure 6—source data 1
Uncropped, labeled western blot.
- https://cdn.elifesciences.org/articles/78945/elife-78945-fig6-data1-v4.zip
Figure 6—figure supplement 1
RT-PCR for additional thermogenic genes.
Bars represent the mean and S.E.M and symbols correspond to each measurement from n=3–4 implants assayed in duplicate.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo sapiens) | Adipose tissue progenitor cell | Corvera Laboratory | Mesenchymal progenitor cells | Cells used in this paper are generated from small fragments of surgically excised male or female human adipose tissue cultured in MatriGel, in the presence of normocin, penicillin and streptomycin. Cultures are recovered using dispase, expanded by two passages using trypsin, and frozen. Cells are then thawed and used for in experiments with no further passaging. Cells are not transformed, nor cultured beyond three passages and are therefore not routinely tested for mycoplasma nor subject to further authentication. Adipose tissue from which the primary cells are derived is obtained in accordance with the UMass Chan Institutional Review Board IRB ID 14734_13 |
| Antibody | Anti-Perilipin-1 (Rabbit monoclonal) | Cell Signalling | #9349 | IF(1:500), |
| Antibody | Anti-MAOA (Rabbit monoclonal) | Cell Signalling | #73030 | IF(1:500) WB (1:500) |
| Antibody | Anti-MAOA (Rabbit monoclonal) | Cell Signalling | #75330 | IF(1:500) WB (1:500) |
| Antibody | Anti-Tyrosine hydroxylase (Rabbit polyclonal) | Millipore Sigma | #AB152 | IF (1:500) |
| Antibody | Anti-CD45 (Mouse monoclonal) | Abcam | #282747 | IF (1:500) |
| Antibody | Anti-F4/80 (Rat monoclonal) | Abcam | #6640 | IF (1:500) |
| Commercial assay, kit | Adiponectin ELISA | Invitrogen | #KHP0041 | Serum diluted 1:50 |
| Other | Isolectin GS-IB4 AlexaFluor-647 conjugate | Invitrogen | #I32450 | Staining, 1:200 |
| Other | Ulex Europaeus Agglutinin I DyLight 594 | Vector Laboratories | #DL-1067–1 | Staining, 1:200 |
| Other | DAPI stain | Invitrogen | #D1306 | Counterstaining 1 µg/mL |
Additional files
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Supplementary file 1
Top 50 Differentially expressed mouse genes between WIM and TIM.
- https://cdn.elifesciences.org/articles/78945/elife-78945-supp1-v4.xlsx
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Supplementary file 2
Differential expression of human genes between WIM and TIM.
- https://cdn.elifesciences.org/articles/78945/elife-78945-supp2-v4.xlsx
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Supplementary file 3
Top 50 genes differentially expressed between non-thermogenic and thermogenic adipocytes prior to implantation.
- https://cdn.elifesciences.org/articles/78945/elife-78945-supp3-v4.xlsx
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Supplementary file 4
Genes differentially expressed between non-thermogenic and thermogenic adipocytes that are maintained differentially expressed in WIM and TIM.
- https://cdn.elifesciences.org/articles/78945/elife-78945-supp4-v4.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/78945/elife-78945-transrepform1-v4.docx
