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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

Analysis of lipid containing structures in live wild type animals and in animals with disrupted W01A8.1 by CARS microscopy.CARS microscopy was performed using constant magnification and intensity settings (20% laser intensity) with the exception of control embryos (C and G) that were examined at 30% laser intensity since the lipid content was lower in wild type. The brightness of the entire figure was digitally enhanced using Adobe Photoshop brightness setting (+150 units) for better visibility of structures. Arrows indicate lipid containing structures in paired panels. (A) The germline of a wild type adult hermaphrodite animal with small lipid containing structures in oocytes and an increase in their number and size during oogenesis. (B) The germline of a mutant hermaphrodite animal. Lipid containing structures are bigger compared to the control animal yet distributed evenly in mutant oocytes. (C) Lipid containing structures localize around the nucleus in one cell wild type embryos. This tendency of the association of the lipid containing structures with nuclear periphery can be seen also during later developmental stages in wild type embryos (D). (E, F) and (H) Enlarged lipid containing structures arranged around the nuclei in mutant embryos. (F) The formation of clusters of lipid containing structures on the periphery of nuclei. (G) Shows a wild type embryo at later stage of the development. (H) The lipid containing structures progressively diminish in size in mutant embryos during later stages of embryonic development (arrowhead). (I) and (J) Lipid containing structures in enterocytes of wild type (I) and mutant (J) animals. In contrast to embryos, which exhibit higher CARS signal and bigger lipid containing structures in mutant animals, gut cells in adult animals show the opposite, that is, a reduced fat-related CARS signal and smaller lipid containing structures in mutant animals. Similarly, lipid containing structures in epidermal cells (K and L) are bigger in wild type animals as shown in the (K) and smaller in mutant animals (L). Bars represent 50 µm.
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fig-5: Analysis of lipid containing structures in live wild type animals and in animals with disrupted W01A8.1 by CARS microscopy.CARS microscopy was performed using constant magnification and intensity settings (20% laser intensity) with the exception of control embryos (C and G) that were examined at 30% laser intensity since the lipid content was lower in wild type. The brightness of the entire figure was digitally enhanced using Adobe Photoshop brightness setting (+150 units) for better visibility of structures. Arrows indicate lipid containing structures in paired panels. (A) The germline of a wild type adult hermaphrodite animal with small lipid containing structures in oocytes and an increase in their number and size during oogenesis. (B) The germline of a mutant hermaphrodite animal. Lipid containing structures are bigger compared to the control animal yet distributed evenly in mutant oocytes. (C) Lipid containing structures localize around the nucleus in one cell wild type embryos. This tendency of the association of the lipid containing structures with nuclear periphery can be seen also during later developmental stages in wild type embryos (D). (E, F) and (H) Enlarged lipid containing structures arranged around the nuclei in mutant embryos. (F) The formation of clusters of lipid containing structures on the periphery of nuclei. (G) Shows a wild type embryo at later stage of the development. (H) The lipid containing structures progressively diminish in size in mutant embryos during later stages of embryonic development (arrowhead). (I) and (J) Lipid containing structures in enterocytes of wild type (I) and mutant (J) animals. In contrast to embryos, which exhibit higher CARS signal and bigger lipid containing structures in mutant animals, gut cells in adult animals show the opposite, that is, a reduced fat-related CARS signal and smaller lipid containing structures in mutant animals. Similarly, lipid containing structures in epidermal cells (K and L) are bigger in wild type animals as shown in the (K) and smaller in mutant animals (L). Bars represent 50 µm.

Mentions: In order to visualize lipid-containing structures in W01A8.1 mutants and in controls in vivo, we used CARS microscopy (done with kind help from Dr. Zhongxiang Jiang, Leica Microsystems, Mannheim, Germany). The CARS systems allow visualization of lipids of specific categories by tuning into symmetric CH2 vibrations of specific fat composing molecules (Zumbusch, Langbein & Borri, 2013). CARS microscopy clearly confirmed the formation of large lipid containing vesicles in early embryos and allowed detailed analysis of the W01A8.1 phenotype. CARS microscopy also confirmed the gradual increase of the size of lipid containing structures during oogenesis (Figs. 5A and 5B), the sudden re-localization of these structures to the periphery of the dividing nucleus in the first embryonic division (Fig. 5E), and the propagation of this phenotype, although with gradually diminishing appearance, throughout embryonic development (Figs. 5F and 5H). CARS microscopy detected this phenomenon also in wild type animals, although the size of lipid containing structures was smaller making the phenomenon of the sudden re-localization of lipid containing structures less obvious (Figs. 5C, 5D and 5G) than in W01A8.1 embryos. In contrast to embryos, lipid-containing structures in intestinal and epidermal cells of adult W01A8.1 mutants (Figs. 5J and 5L) were smaller than lipid-containing structures in control animals (Figs. 5I and 5K).


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)

Analysis of lipid containing structures in live wild type animals and in animals with disrupted W01A8.1 by CARS microscopy.CARS microscopy was performed using constant magnification and intensity settings (20% laser intensity) with the exception of control embryos (C and G) that were examined at 30% laser intensity since the lipid content was lower in wild type. The brightness of the entire figure was digitally enhanced using Adobe Photoshop brightness setting (+150 units) for better visibility of structures. Arrows indicate lipid containing structures in paired panels. (A) The germline of a wild type adult hermaphrodite animal with small lipid containing structures in oocytes and an increase in their number and size during oogenesis. (B) The germline of a mutant hermaphrodite animal. Lipid containing structures are bigger compared to the control animal yet distributed evenly in mutant oocytes. (C) Lipid containing structures localize around the nucleus in one cell wild type embryos. This tendency of the association of the lipid containing structures with nuclear periphery can be seen also during later developmental stages in wild type embryos (D). (E, F) and (H) Enlarged lipid containing structures arranged around the nuclei in mutant embryos. (F) The formation of clusters of lipid containing structures on the periphery of nuclei. (G) Shows a wild type embryo at later stage of the development. (H) The lipid containing structures progressively diminish in size in mutant embryos during later stages of embryonic development (arrowhead). (I) and (J) Lipid containing structures in enterocytes of wild type (I) and mutant (J) animals. In contrast to embryos, which exhibit higher CARS signal and bigger lipid containing structures in mutant animals, gut cells in adult animals show the opposite, that is, a reduced fat-related CARS signal and smaller lipid containing structures in mutant animals. Similarly, lipid containing structures in epidermal cells (K and L) are bigger in wild type animals as shown in the (K) and smaller in mutant animals (L). Bars represent 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig-5: Analysis of lipid containing structures in live wild type animals and in animals with disrupted W01A8.1 by CARS microscopy.CARS microscopy was performed using constant magnification and intensity settings (20% laser intensity) with the exception of control embryos (C and G) that were examined at 30% laser intensity since the lipid content was lower in wild type. The brightness of the entire figure was digitally enhanced using Adobe Photoshop brightness setting (+150 units) for better visibility of structures. Arrows indicate lipid containing structures in paired panels. (A) The germline of a wild type adult hermaphrodite animal with small lipid containing structures in oocytes and an increase in their number and size during oogenesis. (B) The germline of a mutant hermaphrodite animal. Lipid containing structures are bigger compared to the control animal yet distributed evenly in mutant oocytes. (C) Lipid containing structures localize around the nucleus in one cell wild type embryos. This tendency of the association of the lipid containing structures with nuclear periphery can be seen also during later developmental stages in wild type embryos (D). (E, F) and (H) Enlarged lipid containing structures arranged around the nuclei in mutant embryos. (F) The formation of clusters of lipid containing structures on the periphery of nuclei. (G) Shows a wild type embryo at later stage of the development. (H) The lipid containing structures progressively diminish in size in mutant embryos during later stages of embryonic development (arrowhead). (I) and (J) Lipid containing structures in enterocytes of wild type (I) and mutant (J) animals. In contrast to embryos, which exhibit higher CARS signal and bigger lipid containing structures in mutant animals, gut cells in adult animals show the opposite, that is, a reduced fat-related CARS signal and smaller lipid containing structures in mutant animals. Similarly, lipid containing structures in epidermal cells (K and L) are bigger in wild type animals as shown in the (K) and smaller in mutant animals (L). Bars represent 50 µm.
Mentions: In order to visualize lipid-containing structures in W01A8.1 mutants and in controls in vivo, we used CARS microscopy (done with kind help from Dr. Zhongxiang Jiang, Leica Microsystems, Mannheim, Germany). The CARS systems allow visualization of lipids of specific categories by tuning into symmetric CH2 vibrations of specific fat composing molecules (Zumbusch, Langbein & Borri, 2013). CARS microscopy clearly confirmed the formation of large lipid containing vesicles in early embryos and allowed detailed analysis of the W01A8.1 phenotype. CARS microscopy also confirmed the gradual increase of the size of lipid containing structures during oogenesis (Figs. 5A and 5B), the sudden re-localization of these structures to the periphery of the dividing nucleus in the first embryonic division (Fig. 5E), and the propagation of this phenotype, although with gradually diminishing appearance, throughout embryonic development (Figs. 5F and 5H). CARS microscopy detected this phenomenon also in wild type animals, although the size of lipid containing structures was smaller making the phenomenon of the sudden re-localization of lipid containing structures less obvious (Figs. 5C, 5D and 5G) than in W01A8.1 embryos. In contrast to embryos, lipid-containing structures in intestinal and epidermal cells of adult W01A8.1 mutants (Figs. 5J and 5L) were smaller than lipid-containing structures in control animals (Figs. 5I and 5K).

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