J Vet Sci.  2017 Sep;18(3):341-348. 10.4142/jvs.2017.18.3.341.

Guanylyl cyclase C and guanylin reduce fat droplet accumulation in cattle mesenteric adipose tissue

• Yasuda M
• Kawabata J
• Akieda-Asai S
• Nasu T
• Date Y

• KMID: 2412452
• DOI: http://doi.org/10.4142/jvs.2017.18.3.341

Abstract

Guanylyl cyclase C (GC-C) is a member of a family of enzymes that metabolize GTP to cGMP and was first identified as a receptor for heat-stable enterotoxin. Guanylin (GNY) has since been identified as an endogenous ligand for GC-C in the intestine of several mammalian species. The GNY/GC-C system regulates ion transportation and pH in the mucosa. Recently, it was reported that GC-C and GNY are involved in lipid metabolism in rat mesenteric adipose tissue macrophages. To examine the role of GC-C and GNY in lipid metabolism in cattle, we used a bovine mesenteric adipocyte primary culture system and a coculture system for bovine adipocytes and GNY-/GC-C-expressing macrophages. Fat droplets were observed to accumulate in bovine mesenteric adipocytes cultured alone, whereas few fat droplets accumulated in adipocytes indirectly cocultured with macrophages. We also observed that GC-C was present in bovine mesenteric adipose tissue, and that fat droplet accumulation decreased after in vitro GNY administration. Expressions of mRNAs encoding lipogenic factors decreased significantly in adipocytes after either coculture or GNY administration. These results suggest that the GNY/GC-C system is part of the control system for lipid accumulation in bovine mesenteric adipose tissue.

Keyword

cattle; guanylin; guanylyl cyclase C; lipogenic factor; mesenteric adipose tissue

MeSH Terms

Adipocytes/drug effects/metabolism
Adipose Tissue/cytology/*drug effects/metabolism
Animals
Cattle
Coculture Techniques/veterinary
Gastrointestinal Hormones/*pharmacology
Guanylate Cyclase
Lipid Metabolism/drug effects/physiology
Macrophages/drug effects/metabolism
Natriuretic Peptides/*pharmacology
Real-Time Polymerase Chain Reaction/veterinary
Receptors, Enterotoxin/*metabolism
Gastrointestinal Hormones
Natriuretic Peptides
Guanylate Cyclase
Receptors, Enterotoxin

Figure

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  Fig. 1 Expression of guanylyl cyclase C (GC-C) and guanylin (GNY) mRNAs in bovine cultured adipocytes (lane 3), stromal vascular fraction (lane 2), and mesenteric adipose tissue (lane 1).

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  Fig. 2 Oil Red O staining of primary mesenteric adipocytes cultured with guanylyl cyclase C (GC-C)-/guanylin (GNY)-expressing macrophages. (A) Numerous large fat droplets were observed in primary mesenteric adipocytes. (B) The number of fat droplets declined in primary mesenteric adipocytes cocultured with macrophages. Few fat droplets were observed in the primary mesenteric adipocytes. Scale bars = 50 µm.

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  Fig. 3 Oil red O staining of primary mesenteric adipocytes cultured with varying concentrations of rat guanylin (GNY). No GNY (A), 100 nM GNY (B), 500 nM GNY (C), 1 µM GNY (D), or 5 µM GNY (E) added to the culture medium. The number of fat droplets in the primary mesenteric adipocytes decreased with an increased concentration of rat GNY. Scale bars = 50 µm.

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  Fig. 4 Expression of mRNAs encoding various fat-maturing factors in primary mesenteric adipocytes indirectly cocultured with guanylyl cyclase C (GC-C)-/guanylin (GNY)-expressing macrophages. Expression of mRNAs encoding the indicated factors was lower in primary mesenteric adipocytes cocultured with GC-C-/GNY-expressing macrophages. Data are presented as mean ± SEM values. Asterisks denote significant differences (p < 0.05).

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  Fig. 5 Expression of mRNAs encoding various fat-maturing factors in primary mesenteric adipocytes cultured in the presence of various concentrations of rat guanylin (GNY). Expression of mRNAs encoding the indicated factors decreased with an increased concentration of rat GNY. Data are presented as mean ± SEM values. Asterisk denotes a significant difference (p < 0.05 vs. 0 M and 100 nM).

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