Perivascular Adipocytes’ Adipogenesis Is Defined by Their Anatomical Location in the Descending Thoracic Aorta

Figures

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  Figure 1. Adipogenesis and lipogenesis in adipocytes from anterior and lateral thoracic aortic perivascular adipose tissue (PVAT) strips. (A) Low magnification representative image of the rat’s thoracic aorta with its three strips of PVAT, one anterior and two lateral. (B) Representative images of adipocyte progenitors and adipocytes at 4 and 7 days post-induction (n = 7). Lipid droplets were stained with Bodipy (Thermo Fisher; green) and nuclei with NucRed (Biotium; red). Scale bar = 400 µm. (C) Adipogenic efficiency as calculated by the IncuCyte imager (Sartorius; number of cells with one or more lipid droplets over total number of cells per well); n = 7. (D) Neutral lipid accumulation and triglycerides content (using Triglyceride-Glo Assay) relative to Anterior after 7 d in culture; n = 7. (E) Adiponectin content in media and cells as measured by ELISA (Thermo Fisher) after 7 d in culture; n = 4. Dots indicate individual data points. Error bars indicate SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

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  Figure 2. Adipocyte progenitors and adipocytes from the thoracic aorta’s anterior and lateral PVAT exhibit different transcriptomic profiles. Adipose tissue samples were collected from anterior (AP) and lateral (LP) PVAT of SD male rats (n = 3). (A). Principal component (3D) and clustering analyses show gene transcription comparisons between adipocyte progenitors and mature adipocytes in AP and LP. (B). Volcano plots of differentially expressed genes in adipocyte progenitors and adipocytes from LP when compared to AP. Overall results of FPKM cluster analysis from LP and AP adipocyte progenitors and adipocytes (using log2 (FPKM + 1) values). Red = high expression levels; green = low expression levels. Colors ranging from red to green indicate that log2 (FPKM + 1) values range from large to small. (C). Heatmap displays gene expression levels, with rows representing genes and columns representing samples (n = 3 in each group). Color intensity indicates z-score expression magnitude, from low (cyan) to high (yellow). (D). Volcano plots of differentially expressed genes in LP and AP adipocyte progenitors when compared to adipocytes from the same site. (E). Fragments per kilobase per million…

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  Figure 3. Adipogenesis Alters Extracellular Matrix Gene Transcription Patterns in Thoracic Aorta AP and LP Adipocyte Progenitor Cells. Adipose tissue samples were collected from anterior (AP) and lateral (LP) PVAT of SD male rats (n = 3). (A) Results of hypergeometric tests showing collagen and integrin gene family enrichment among differentially expressed genes in progenitors vs. adipocytes (top, gray) and anterior vs. lateral (bottom, blue) comparisons. The vertical red line indicates p = 0.05. Fragments per kilobase per million mapped reads (FPKM) of collagens and thrombospondin 1 and integrins and fibronectin 1 (B) detected in adipocyte progenitors from AP and LP and adipocytes after 7 days of adipogenesis induction. (C) Collagens and integrins were differentially expressed in both adipocyte progenitors and adipocytes from AP and LP. Data are means ± SEM. Comparison lines with * differ (p < 0.05).

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  Figure 4. Adipogenesis in Thoracic Aorta AP and LP Adipocyte Progenitor Cells. Adipose tissue samples were collected from anterior (AP) and lateral (LP) PVAT of SD rats (n = 4). (A). Adipogenesis efficiency was examined using the IncuCyte imager (Sartorius; number of cells with one or more lipid droplets over total number of cells per well); n = 4. (B). Triglyceride content in cells were measured using Triglyceride-Glo Assay; n = 4. (C). Adiponectin secretion content from cells measured using ELISA (Thermo Fisher); n = 4. Dots in graphs are individual data points and error bars indicate the SEM. Bars with different superscripts a,b are different (p < 0.05).