Limits...
Perilipin-related protein regulates lipid metabolism in C. elegans.

Chughtai AA, Kaššák F, Kostrouchová M, Novotný JP, Krause MW, Saudek V, Kostrouch Z, Kostrouchová M - PeerJ (2015)

Bottom Line: Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes.In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals.Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles University in Prague , Albertov, Prague , Czech Republic.

ABSTRACT
Perilipins are lipid droplet surface proteins that contribute to fat metabolism by controlling the access of lipids to lipolytic enzymes. Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes. Here we identify the protein encoded by the W01A8.1 gene in Caenorhabditis elegans as the closest homologue and likely orthologue of metazoan perilipin. We demonstrate that nematode W01A8.1 is a cytoplasmic protein residing on lipid droplets similarly as human perilipins 1 and 2. Downregulation or elimination of W01A8.1 affects the appearance of lipid droplets resulting in the formation of large lipid droplets localized around the dividing nucleus during the early zygotic divisions. Visualization of lipid containing structures by CARS microscopy in vivo showed that lipid-containing structures become gradually enlarged during oogenesis and relocate during the first zygotic division around the dividing nucleus. In mutant embryos, the lipid containing structures show defective intracellular distribution in subsequent embryonic divisions and become gradually smaller during further development. In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals. Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

No MeSH data available.


Related in: MedlinePlus

Expression of human perilipins fused to GFP in C. elegans.(A–C) Expression of human PLIN1::GFP in live transgenic C. elegans. PLIN1::GFP is localized on vesicles with an appearance of lipid droplets. PLIN2::GFP (D–F) is localized in transgenic animals on vesicular structures with an appearance of lipid droplets similarly as PLIN1::GFP. PLIN3::GFP (G, H and I) yields a more diffuse cytoplasmic pattern with faintly stained vesicular structures. (A, D) and (G) and details in (C, F) and (I) show GFP in fluorescence microscopy and (B, E) and (H) corresponding areas to (A, D) and (G) in Nomarski optics. (J, K) and (L) show PLIN2::GFP in fluorescence microscopy (J) in fixed C. elegans stained with LipidTox (K). The area indicated by the white rectangle in (J) and (K) is magnified and merged for co-localization of PLIN2::GFP (green) and LipidTox (red) in (L). Arrows indicate lipid droplets clearly marked by GFP with the LipidTox positive content. Bars represent 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4562238&req=5

fig-3: Expression of human perilipins fused to GFP in C. elegans.(A–C) Expression of human PLIN1::GFP in live transgenic C. elegans. PLIN1::GFP is localized on vesicles with an appearance of lipid droplets. PLIN2::GFP (D–F) is localized in transgenic animals on vesicular structures with an appearance of lipid droplets similarly as PLIN1::GFP. PLIN3::GFP (G, H and I) yields a more diffuse cytoplasmic pattern with faintly stained vesicular structures. (A, D) and (G) and details in (C, F) and (I) show GFP in fluorescence microscopy and (B, E) and (H) corresponding areas to (A, D) and (G) in Nomarski optics. (J, K) and (L) show PLIN2::GFP in fluorescence microscopy (J) in fixed C. elegans stained with LipidTox (K). The area indicated by the white rectangle in (J) and (K) is magnified and merged for co-localization of PLIN2::GFP (green) and LipidTox (red) in (L). Arrows indicate lipid droplets clearly marked by GFP with the LipidTox positive content. Bars represent 50 µm.

Mentions: We prepared transgenic C. elegans lines expressing human PLIN1, PLIN2 and PLIN3 fused to GFP and regulated by the W01A8.1 promoter. PLIN1::GFP and PLIN2::GFP were localized on spherical cytoplasmic structures primarily in gut and epidermal cells (Figs. 3A, 3C, 3D and 3F) with identical appearance as W01A8.1 translational reporter GFP tagged proteins and Drosophila PLIN1::GFP expressed in C. elegans as reported by Liu et al. (2014). PLIN3 expression was diffusely cytoplasmic and only faintly defined spherical structures (Figs. 3G and 3I). The structures clearly labeled with PLIN1::GFP and PLIN2::GFP were also positive in LipidTox staining (shown for PLIN2::GFP in Figs. 3J–3L). We conclude that W01A8.1 proteins are localized on the same structures as human PLIN1 and PLIN2.


Perilipin-related protein regulates lipid metabolism in C. elegans.

Chughtai AA, Kaššák F, Kostrouchová M, Novotný JP, Krause MW, Saudek V, Kostrouch Z, Kostrouchová M - PeerJ (2015)

Expression of human perilipins fused to GFP in C. elegans.(A–C) Expression of human PLIN1::GFP in live transgenic C. elegans. PLIN1::GFP is localized on vesicles with an appearance of lipid droplets. PLIN2::GFP (D–F) is localized in transgenic animals on vesicular structures with an appearance of lipid droplets similarly as PLIN1::GFP. PLIN3::GFP (G, H and I) yields a more diffuse cytoplasmic pattern with faintly stained vesicular structures. (A, D) and (G) and details in (C, F) and (I) show GFP in fluorescence microscopy and (B, E) and (H) corresponding areas to (A, D) and (G) in Nomarski optics. (J, K) and (L) show PLIN2::GFP in fluorescence microscopy (J) in fixed C. elegans stained with LipidTox (K). The area indicated by the white rectangle in (J) and (K) is magnified and merged for co-localization of PLIN2::GFP (green) and LipidTox (red) in (L). Arrows indicate lipid droplets clearly marked by GFP with the LipidTox positive content. Bars represent 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4562238&req=5

fig-3: Expression of human perilipins fused to GFP in C. elegans.(A–C) Expression of human PLIN1::GFP in live transgenic C. elegans. PLIN1::GFP is localized on vesicles with an appearance of lipid droplets. PLIN2::GFP (D–F) is localized in transgenic animals on vesicular structures with an appearance of lipid droplets similarly as PLIN1::GFP. PLIN3::GFP (G, H and I) yields a more diffuse cytoplasmic pattern with faintly stained vesicular structures. (A, D) and (G) and details in (C, F) and (I) show GFP in fluorescence microscopy and (B, E) and (H) corresponding areas to (A, D) and (G) in Nomarski optics. (J, K) and (L) show PLIN2::GFP in fluorescence microscopy (J) in fixed C. elegans stained with LipidTox (K). The area indicated by the white rectangle in (J) and (K) is magnified and merged for co-localization of PLIN2::GFP (green) and LipidTox (red) in (L). Arrows indicate lipid droplets clearly marked by GFP with the LipidTox positive content. Bars represent 50 µm.
Mentions: We prepared transgenic C. elegans lines expressing human PLIN1, PLIN2 and PLIN3 fused to GFP and regulated by the W01A8.1 promoter. PLIN1::GFP and PLIN2::GFP were localized on spherical cytoplasmic structures primarily in gut and epidermal cells (Figs. 3A, 3C, 3D and 3F) with identical appearance as W01A8.1 translational reporter GFP tagged proteins and Drosophila PLIN1::GFP expressed in C. elegans as reported by Liu et al. (2014). PLIN3 expression was diffusely cytoplasmic and only faintly defined spherical structures (Figs. 3G and 3I). The structures clearly labeled with PLIN1::GFP and PLIN2::GFP were also positive in LipidTox staining (shown for PLIN2::GFP in Figs. 3J–3L). We conclude that W01A8.1 proteins are localized on the same structures as human PLIN1 and PLIN2.

Bottom Line: Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes.In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals.Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles University in Prague , Albertov, Prague , Czech Republic.

ABSTRACT
Perilipins are lipid droplet surface proteins that contribute to fat metabolism by controlling the access of lipids to lipolytic enzymes. Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes. Here we identify the protein encoded by the W01A8.1 gene in Caenorhabditis elegans as the closest homologue and likely orthologue of metazoan perilipin. We demonstrate that nematode W01A8.1 is a cytoplasmic protein residing on lipid droplets similarly as human perilipins 1 and 2. Downregulation or elimination of W01A8.1 affects the appearance of lipid droplets resulting in the formation of large lipid droplets localized around the dividing nucleus during the early zygotic divisions. Visualization of lipid containing structures by CARS microscopy in vivo showed that lipid-containing structures become gradually enlarged during oogenesis and relocate during the first zygotic division around the dividing nucleus. In mutant embryos, the lipid containing structures show defective intracellular distribution in subsequent embryonic divisions and become gradually smaller during further development. In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals. Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

No MeSH data available.


Related in: MedlinePlus