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Perilipin-2 Modulates Lipid Absorption and Microbiome Responses in the Mouse Intestine.

Frank DN, Bales ES, Monks J, Jackman MJ, MacLean PS, Ir D, Robertson CE, Orlicky DJ, McManaman JL - PLoS ONE (2015)

Bottom Line: Here we test the hypotheses that Plin2 function impacts the earliest steps of HF diet-mediated pathogenesis as well as the dynamics of diet-associated changes in gut microbiome diversity and function.Plin2- mice had significantly lower respiratory exchange ratios, diminished frequencies of enterocyte CLDs, and increased fecal triglyceride levels compared with WT mice.Microbiome analyses, employing both 16S rRNA profiling and metagenomic deep sequencing, indicated that dietary fat content and Plin2 genotype were significantly and independently associated with gut microbiome composition, diversity, and functional differences.

View Article: PubMed Central - PubMed

Affiliation: Division of Infectious Disease, University of Colorado School of Medicine, Aurora, Colorado, United States of America; Microbiome Research Consortium, University of Colorado School of Medicine, Aurora, Colorado, United States of America.

ABSTRACT
Obesity and its co-morbidities, such as fatty liver disease, are increasingly prevalent worldwide health problems. Intestinal microorganisms have emerged as critical factors linking diet to host physiology and metabolic function, particularly in the context of lipid homeostasis. We previously demonstrated that deletion of the cytoplasmic lipid drop (CLD) protein Perilipin-2 (Plin2) in mice largely abrogates long-term deleterious effects of a high fat (HF) diet. Here we test the hypotheses that Plin2 function impacts the earliest steps of HF diet-mediated pathogenesis as well as the dynamics of diet-associated changes in gut microbiome diversity and function. WT and perilipin-2 mice raised on a standard chow diet were randomized to either low fat (LF) or HF diets. After four days, animals were assessed for changes in physiological (body weight, energy balance, and fecal triglyceride levels), histochemical (enterocyte CLD content), and fecal microbiome parameters. Plin2- mice had significantly lower respiratory exchange ratios, diminished frequencies of enterocyte CLDs, and increased fecal triglyceride levels compared with WT mice. Microbiome analyses, employing both 16S rRNA profiling and metagenomic deep sequencing, indicated that dietary fat content and Plin2 genotype were significantly and independently associated with gut microbiome composition, diversity, and functional differences. These data demonstrate that Plin2 modulates rapid effects of diet on fecal lipid levels, enterocyte CLD contents, and fuel utilization properties of mice that correlate with structural and functional differences in their gut microbial communities. Collectively, the data provide evidence of Plin2 regulated intestinal lipid uptake, which contributes to rapid changes in the gut microbial communities implicated in diet-induced obesity.

No MeSH data available.


Related in: MedlinePlus

Effects of HF diet feeding on intestinal lipid properties.(A) Representative H&E stained jejunal sections from HF fed WT and Plin2- mice. Note the apparent greater abundance of CLD (arrows) in intestinal villi from WT mice compared to Plin2- mice. Scale bar = 100 μm. (B) Representative immunofluorescence images of jejunal sections from fasted and HF fed WT and Plin2- mice stained with guinea pig anti-Plin2 (green) and rabbit anti-Plin3 (red) showing Plin2- (green arrows) and Plin3- (red arrows) coated CLD. Yellow arrow indicates CLD coated with Plin2 and Plin3. (C) BODIPY stained CLD (Red) and Alexafluor-WGA (green) stained intestinal villi surfaces from fasted and fed WT and Plin2- mice. Scale bar = 100 μm. (D) Relative enterocyte lipid levels. Values are mean BODIPY fluorescence area/cell ± SEM from 8–10 200X sections from the jejunal and ileal regions of each mouse. Four animals per group were analyzed. P-values were determined using the unpaired t-test (Prism 6, GraphPad Prism).
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pone.0131944.g003: Effects of HF diet feeding on intestinal lipid properties.(A) Representative H&E stained jejunal sections from HF fed WT and Plin2- mice. Note the apparent greater abundance of CLD (arrows) in intestinal villi from WT mice compared to Plin2- mice. Scale bar = 100 μm. (B) Representative immunofluorescence images of jejunal sections from fasted and HF fed WT and Plin2- mice stained with guinea pig anti-Plin2 (green) and rabbit anti-Plin3 (red) showing Plin2- (green arrows) and Plin3- (red arrows) coated CLD. Yellow arrow indicates CLD coated with Plin2 and Plin3. (C) BODIPY stained CLD (Red) and Alexafluor-WGA (green) stained intestinal villi surfaces from fasted and fed WT and Plin2- mice. Scale bar = 100 μm. (D) Relative enterocyte lipid levels. Values are mean BODIPY fluorescence area/cell ± SEM from 8–10 200X sections from the jejunal and ileal regions of each mouse. Four animals per group were analyzed. P-values were determined using the unpaired t-test (Prism 6, GraphPad Prism).

Mentions: The effects of Plin2 deletion on intestinal lipid properties were evaluated by comparing intestinal histology and intestinal lipid droplet staining properties in 4 day HF fed WT and Plin2- mice that were fasted and then refed a HF diet. Significant numbers of large and small CLD were detected in enterocytes of H&E stained sections of WT mice. In contrast, only limited numbers of CLD were found in jejunal enterocytes of Plin2- mice (Fig 3A). Immunofluorescence analysis documented that Plin2 and Plin3 coated CLD in WT enterocytes, and in some cases both Plin2 and Plin3 were found to coat the same lipid droplet (Fig 3B). As expected, enterocyte CLD in Plin2- mice were positive for Plin3 but lacked Plin2 (Fig 3B). To quantify effects of Plin2 deletion on enterocyte lipid content, we stained intestinal sections from the jejunum and ilium of fasted and refed mice with BODIPY to label neutral lipids (Fig 3C) and calculated the relative level of fluorescence staining per cell (Fig 3D) [22]. The BODIPY fluorescence results indicate that neutral lipid levels in enterocytes in jejunum and ileum of WT mice are significantly greater than those of Plin2- mice. Collectively, the results from quantitation of fecal and enterocyte lipid levels are consistent with the proposed role of Plin2 in regulating intestinal lipid absorption after high fat feeding [15], and provide evidence that Plin2 loss may impair this process. Moreover, the observation that Plin3 coats CLD in enterocytes of Plin2- mice combined with the reduced numbers of BODIPY-stained CLD in jejunal and ileal enterocytes of Plin2- mice, suggests that Plin3 may not fully compensate for Plin2 in regulating intestinal lipid properties.


Perilipin-2 Modulates Lipid Absorption and Microbiome Responses in the Mouse Intestine.

Frank DN, Bales ES, Monks J, Jackman MJ, MacLean PS, Ir D, Robertson CE, Orlicky DJ, McManaman JL - PLoS ONE (2015)

Effects of HF diet feeding on intestinal lipid properties.(A) Representative H&E stained jejunal sections from HF fed WT and Plin2- mice. Note the apparent greater abundance of CLD (arrows) in intestinal villi from WT mice compared to Plin2- mice. Scale bar = 100 μm. (B) Representative immunofluorescence images of jejunal sections from fasted and HF fed WT and Plin2- mice stained with guinea pig anti-Plin2 (green) and rabbit anti-Plin3 (red) showing Plin2- (green arrows) and Plin3- (red arrows) coated CLD. Yellow arrow indicates CLD coated with Plin2 and Plin3. (C) BODIPY stained CLD (Red) and Alexafluor-WGA (green) stained intestinal villi surfaces from fasted and fed WT and Plin2- mice. Scale bar = 100 μm. (D) Relative enterocyte lipid levels. Values are mean BODIPY fluorescence area/cell ± SEM from 8–10 200X sections from the jejunal and ileal regions of each mouse. Four animals per group were analyzed. P-values were determined using the unpaired t-test (Prism 6, GraphPad Prism).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493139&req=5

pone.0131944.g003: Effects of HF diet feeding on intestinal lipid properties.(A) Representative H&E stained jejunal sections from HF fed WT and Plin2- mice. Note the apparent greater abundance of CLD (arrows) in intestinal villi from WT mice compared to Plin2- mice. Scale bar = 100 μm. (B) Representative immunofluorescence images of jejunal sections from fasted and HF fed WT and Plin2- mice stained with guinea pig anti-Plin2 (green) and rabbit anti-Plin3 (red) showing Plin2- (green arrows) and Plin3- (red arrows) coated CLD. Yellow arrow indicates CLD coated with Plin2 and Plin3. (C) BODIPY stained CLD (Red) and Alexafluor-WGA (green) stained intestinal villi surfaces from fasted and fed WT and Plin2- mice. Scale bar = 100 μm. (D) Relative enterocyte lipid levels. Values are mean BODIPY fluorescence area/cell ± SEM from 8–10 200X sections from the jejunal and ileal regions of each mouse. Four animals per group were analyzed. P-values were determined using the unpaired t-test (Prism 6, GraphPad Prism).
Mentions: The effects of Plin2 deletion on intestinal lipid properties were evaluated by comparing intestinal histology and intestinal lipid droplet staining properties in 4 day HF fed WT and Plin2- mice that were fasted and then refed a HF diet. Significant numbers of large and small CLD were detected in enterocytes of H&E stained sections of WT mice. In contrast, only limited numbers of CLD were found in jejunal enterocytes of Plin2- mice (Fig 3A). Immunofluorescence analysis documented that Plin2 and Plin3 coated CLD in WT enterocytes, and in some cases both Plin2 and Plin3 were found to coat the same lipid droplet (Fig 3B). As expected, enterocyte CLD in Plin2- mice were positive for Plin3 but lacked Plin2 (Fig 3B). To quantify effects of Plin2 deletion on enterocyte lipid content, we stained intestinal sections from the jejunum and ilium of fasted and refed mice with BODIPY to label neutral lipids (Fig 3C) and calculated the relative level of fluorescence staining per cell (Fig 3D) [22]. The BODIPY fluorescence results indicate that neutral lipid levels in enterocytes in jejunum and ileum of WT mice are significantly greater than those of Plin2- mice. Collectively, the results from quantitation of fecal and enterocyte lipid levels are consistent with the proposed role of Plin2 in regulating intestinal lipid absorption after high fat feeding [15], and provide evidence that Plin2 loss may impair this process. Moreover, the observation that Plin3 coats CLD in enterocytes of Plin2- mice combined with the reduced numbers of BODIPY-stained CLD in jejunal and ileal enterocytes of Plin2- mice, suggests that Plin3 may not fully compensate for Plin2 in regulating intestinal lipid properties.

Bottom Line: Here we test the hypotheses that Plin2 function impacts the earliest steps of HF diet-mediated pathogenesis as well as the dynamics of diet-associated changes in gut microbiome diversity and function.Plin2- mice had significantly lower respiratory exchange ratios, diminished frequencies of enterocyte CLDs, and increased fecal triglyceride levels compared with WT mice.Microbiome analyses, employing both 16S rRNA profiling and metagenomic deep sequencing, indicated that dietary fat content and Plin2 genotype were significantly and independently associated with gut microbiome composition, diversity, and functional differences.

View Article: PubMed Central - PubMed

Affiliation: Division of Infectious Disease, University of Colorado School of Medicine, Aurora, Colorado, United States of America; Microbiome Research Consortium, University of Colorado School of Medicine, Aurora, Colorado, United States of America.

ABSTRACT
Obesity and its co-morbidities, such as fatty liver disease, are increasingly prevalent worldwide health problems. Intestinal microorganisms have emerged as critical factors linking diet to host physiology and metabolic function, particularly in the context of lipid homeostasis. We previously demonstrated that deletion of the cytoplasmic lipid drop (CLD) protein Perilipin-2 (Plin2) in mice largely abrogates long-term deleterious effects of a high fat (HF) diet. Here we test the hypotheses that Plin2 function impacts the earliest steps of HF diet-mediated pathogenesis as well as the dynamics of diet-associated changes in gut microbiome diversity and function. WT and perilipin-2 mice raised on a standard chow diet were randomized to either low fat (LF) or HF diets. After four days, animals were assessed for changes in physiological (body weight, energy balance, and fecal triglyceride levels), histochemical (enterocyte CLD content), and fecal microbiome parameters. Plin2- mice had significantly lower respiratory exchange ratios, diminished frequencies of enterocyte CLDs, and increased fecal triglyceride levels compared with WT mice. Microbiome analyses, employing both 16S rRNA profiling and metagenomic deep sequencing, indicated that dietary fat content and Plin2 genotype were significantly and independently associated with gut microbiome composition, diversity, and functional differences. These data demonstrate that Plin2 modulates rapid effects of diet on fecal lipid levels, enterocyte CLD contents, and fuel utilization properties of mice that correlate with structural and functional differences in their gut microbial communities. Collectively, the data provide evidence of Plin2 regulated intestinal lipid uptake, which contributes to rapid changes in the gut microbial communities implicated in diet-induced obesity.

No MeSH data available.


Related in: MedlinePlus