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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

Loss of W01A8.1 function results in abnormal lipid droplet appearance.(A) and (B) An embryo from a hermaphrodite inhibited for W01A8.1 function by RNAi. Large lipid droplets stained by LipidTox (B) are visible also in Nomarski optics (A) in contrast with a control embryo that has only small and more evenly distributed lipid droplets (C—Nomarski optics and D—LipidTox staining). (E–J, L) and (M) Images of structures observed in animals with disrupted W01A8.1. (E) and (F) show structures with the appearance of lipid droplets in the intestine of an animal with disrupted W01A8.1 balanced with the synthetic transgene W01A8.1(synth)::gfp. GFP tagged synthetic W01A8.1a is localized on lipid droplets-like vesicular structures (E). (F) Shows the same area stained with LipidTox. (G) Shows in magnification a merged image of the area indicated by white rectangles in (E) and (F). Arrows indicate W01A8.1(synth)::GFP labeled lipid droplets (green) positive for lipids in LipidTox staining (red). (H) and (I) show an embryo of a parent with disrupted W01A8.1 that had confirmed loss of the balancing transgene. Large LipidTox stained droplets are visible in Nomarski optics (H) as well as in LipidTox staining (I). Their enlargement is clearly visible in comparison with the wild type embryo shown in (C) and (D). (J) and (K) are images of live animals. (J) Shows an embryo with disrupted W01A8.1 and confirmed loss of the balancing transgene. Large vesicular structures are formed around the dividing nucleus (arrows). (K) Shows a control embryo with normal appearance of the nuclear periphery (arrow). (L) and (M) show a one cell embryo from a parent with disrupted W01A8.1 and confirmed loss of extrachromosomal array containing W01A8.1(a)synth::gfp after fixation and staining by LipidTox with large lipid droplets around the dividing nucleus visible in Nomarski optics (L) and positive for lipids in LipidTox staing (M) indicated by arrows. Bars represent 10 µm.
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fig-4: Loss of W01A8.1 function results in abnormal lipid droplet appearance.(A) and (B) An embryo from a hermaphrodite inhibited for W01A8.1 function by RNAi. Large lipid droplets stained by LipidTox (B) are visible also in Nomarski optics (A) in contrast with a control embryo that has only small and more evenly distributed lipid droplets (C—Nomarski optics and D—LipidTox staining). (E–J, L) and (M) Images of structures observed in animals with disrupted W01A8.1. (E) and (F) show structures with the appearance of lipid droplets in the intestine of an animal with disrupted W01A8.1 balanced with the synthetic transgene W01A8.1(synth)::gfp. GFP tagged synthetic W01A8.1a is localized on lipid droplets-like vesicular structures (E). (F) Shows the same area stained with LipidTox. (G) Shows in magnification a merged image of the area indicated by white rectangles in (E) and (F). Arrows indicate W01A8.1(synth)::GFP labeled lipid droplets (green) positive for lipids in LipidTox staining (red). (H) and (I) show an embryo of a parent with disrupted W01A8.1 that had confirmed loss of the balancing transgene. Large LipidTox stained droplets are visible in Nomarski optics (H) as well as in LipidTox staining (I). Their enlargement is clearly visible in comparison with the wild type embryo shown in (C) and (D). (J) and (K) are images of live animals. (J) Shows an embryo with disrupted W01A8.1 and confirmed loss of the balancing transgene. Large vesicular structures are formed around the dividing nucleus (arrows). (K) Shows a control embryo with normal appearance of the nuclear periphery (arrow). (L) and (M) show a one cell embryo from a parent with disrupted W01A8.1 and confirmed loss of extrachromosomal array containing W01A8.1(a)synth::gfp after fixation and staining by LipidTox with large lipid droplets around the dividing nucleus visible in Nomarski optics (L) and positive for lipids in LipidTox staing (M) indicated by arrows. Bars represent 10 µm.

Mentions: Staining of adult hermaphrodites with LipidTox (after formaldehyde fixation) revealed larger lipid droplets in early embryos derived from adults inhibited for W01A8.1 (Figs. 4A and 4B) compared to controls (Figs. 4C and 4D).


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)

Loss of W01A8.1 function results in abnormal lipid droplet appearance.(A) and (B) An embryo from a hermaphrodite inhibited for W01A8.1 function by RNAi. Large lipid droplets stained by LipidTox (B) are visible also in Nomarski optics (A) in contrast with a control embryo that has only small and more evenly distributed lipid droplets (C—Nomarski optics and D—LipidTox staining). (E–J, L) and (M) Images of structures observed in animals with disrupted W01A8.1. (E) and (F) show structures with the appearance of lipid droplets in the intestine of an animal with disrupted W01A8.1 balanced with the synthetic transgene W01A8.1(synth)::gfp. GFP tagged synthetic W01A8.1a is localized on lipid droplets-like vesicular structures (E). (F) Shows the same area stained with LipidTox. (G) Shows in magnification a merged image of the area indicated by white rectangles in (E) and (F). Arrows indicate W01A8.1(synth)::GFP labeled lipid droplets (green) positive for lipids in LipidTox staining (red). (H) and (I) show an embryo of a parent with disrupted W01A8.1 that had confirmed loss of the balancing transgene. Large LipidTox stained droplets are visible in Nomarski optics (H) as well as in LipidTox staining (I). Their enlargement is clearly visible in comparison with the wild type embryo shown in (C) and (D). (J) and (K) are images of live animals. (J) Shows an embryo with disrupted W01A8.1 and confirmed loss of the balancing transgene. Large vesicular structures are formed around the dividing nucleus (arrows). (K) Shows a control embryo with normal appearance of the nuclear periphery (arrow). (L) and (M) show a one cell embryo from a parent with disrupted W01A8.1 and confirmed loss of extrachromosomal array containing W01A8.1(a)synth::gfp after fixation and staining by LipidTox with large lipid droplets around the dividing nucleus visible in Nomarski optics (L) and positive for lipids in LipidTox staing (M) indicated by arrows. Bars represent 10 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig-4: Loss of W01A8.1 function results in abnormal lipid droplet appearance.(A) and (B) An embryo from a hermaphrodite inhibited for W01A8.1 function by RNAi. Large lipid droplets stained by LipidTox (B) are visible also in Nomarski optics (A) in contrast with a control embryo that has only small and more evenly distributed lipid droplets (C—Nomarski optics and D—LipidTox staining). (E–J, L) and (M) Images of structures observed in animals with disrupted W01A8.1. (E) and (F) show structures with the appearance of lipid droplets in the intestine of an animal with disrupted W01A8.1 balanced with the synthetic transgene W01A8.1(synth)::gfp. GFP tagged synthetic W01A8.1a is localized on lipid droplets-like vesicular structures (E). (F) Shows the same area stained with LipidTox. (G) Shows in magnification a merged image of the area indicated by white rectangles in (E) and (F). Arrows indicate W01A8.1(synth)::GFP labeled lipid droplets (green) positive for lipids in LipidTox staining (red). (H) and (I) show an embryo of a parent with disrupted W01A8.1 that had confirmed loss of the balancing transgene. Large LipidTox stained droplets are visible in Nomarski optics (H) as well as in LipidTox staining (I). Their enlargement is clearly visible in comparison with the wild type embryo shown in (C) and (D). (J) and (K) are images of live animals. (J) Shows an embryo with disrupted W01A8.1 and confirmed loss of the balancing transgene. Large vesicular structures are formed around the dividing nucleus (arrows). (K) Shows a control embryo with normal appearance of the nuclear periphery (arrow). (L) and (M) show a one cell embryo from a parent with disrupted W01A8.1 and confirmed loss of extrachromosomal array containing W01A8.1(a)synth::gfp after fixation and staining by LipidTox with large lipid droplets around the dividing nucleus visible in Nomarski optics (L) and positive for lipids in LipidTox staing (M) indicated by arrows. Bars represent 10 µm.
Mentions: Staining of adult hermaphrodites with LipidTox (after formaldehyde fixation) revealed larger lipid droplets in early embryos derived from adults inhibited for W01A8.1 (Figs. 4A and 4B) compared to controls (Figs. 4C and 4D).

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