Stearoyl-coenzyme A desaturase 1 is required for lipid droplet formation in pig embryo

in Reproduction
Correspondence should be addressed to C-K Lee; Email: leeck@snu.ac.kr
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Lipid droplets (LD) provide a source of energy, and their importance during embryogenesis has been increasingly recognized. In particular, pig embryos have larger amounts of intercellular lipid bilayers than other mammalian species, suggesting that porcine embryos are more dependent on lipid metabolic pathways. The objective of the present study was to detect the effect of stearoyl-coenzyme A desaturase 1 (SCD1) on LD formation and to associate these effects with the mRNA abundance of LD formation-related genes (SREBP, ARF1, COPG2, PLD1 and ERK2) in in vitro-produced porcine embryos. To determine the effect of SCD1 on LD formation and related genes, we examined the effects of SCD1 inhibition using CAY10566 (an SCD1 inhibitor, 50 μM) on parthenogenetic embryos. SCD1 inhibition downregulated the mRNA levels of LD formation-related genes and embryo development. Our results revealed that SCD1 functions in the regulation of LD formation via phospholipid formation and embryo development. In addition, we treated parthenogenetic embryos with oleic acid (100 μM), which led to a significant increase in the blastocyst formation rate, LD size and number compared to controls. Remarkably, the adverse effects of the SCD1 inhibitor could be counteracted by oleic acid. These data suggest that porcine embryos can use exogenous oleic acid as a metabolic energy source.

 

    Society for Reproduction and Fertility

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    Changes in the SCD1 gene during porcine embryonic stages. Immunocytochemistry for SCD1 expression in different stages of in vitro-produced porcine embryos. The nuclei of embryos were counterstained with Hoechst 33342 (orange) (A). Real-time quantification of SCD1 expression in different embryonic stages (B). Different superscript letters indicate significant differences (P < 0.05). Scale bar: 100 µm.

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    Effect of CAY10566 (SCD1 inhibitor) and oleic acid on embryos. Percentage of parthenogenetic embryo blastocyst rate were counted at day 7 of culture from embryos exposed to CAY10566, oleic acid (OA) and CAY10566 with OA treated simultaneously (168 h). Dose-dependent effect of oleic acid on blastocyst formation rate (A, B and C). Added 100 μM of OA group showed an increase of embryo development rate without toxicity (A and B), but toxicity was confirmed at higher (250–500 μM) concentrations (C). Dose-dependent effect of CAY10566 on blastocyst formation rate (D). Effect of oleic acid on CAY10566-treated parthenogenetic embryo blastocyst rate. Low concentration of CAY10566 has no effect on blastocyst rate (1–25 µM, data not shown) (E). Protein levels of the SCD1 gene in each embryo stage (2C; 2cell, 4C; 4cell, 6C; 6cell, Mo; Morulae) (F) and mean fluorescence intensity of SCD1 (G). Different superscript letters indicate significant differences between the control (con), DMSO (vehicle control, V.con), CAY10566-treated (CAY) and CAY10566 + oleic acid (CAY + OA) groups (P < 0.05). DMSO was used only in CAY and OA was in liquid form. Scale bar: 100 µm. Each group has 3–4 replicates.

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    Transcription levels of lipid droplet formation-related genes. Transcription levels of lipid droplet formation-related genes shown for control SCD1-inhibited blastocysts (CAY10566) (A), oleic acid-treated group (B) and CAY10566 with oleic acid-treated blastocysts (C). Each group of blastocysts was pulled at day 7 after parthenogenesis. Each group has three replicates. Different superscript letters indicate significant differences between the control (CON), CAY10566-treated (CAY), oleic acid-treated (OA) and CAY10566 + oleic acid (CAY + O) groups (P < 0.05).

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    Lipid droplet formation of CAY10566- and oleic acid-treated embryos. Porcine embryos at different stages stained with Hoechst 33342, fixed and observed under z-stack of confocal microscopy. Nile Red fluorescence indicating lipid droplets (A). Stage-dependent effect of CAY10566 and oleic acid on lipid droplet number (B). Lipid droplet size was calculated from (C). Different superscript letters indicate significant differences between the control (CON), oleic acid- (OA), CAY10566- (CAY) and CAY10566 + oleic acid-treated groups (CAY + OA) (P < 0.05). Scale bar: 50 µm.

References

  • AardemaHVosPLLolicatoFRoelenBAKnijnHMVaandragerABHelmsJBGadellaBM 2011 Oleic acid prevents detrimental effects of saturated fatty acids on bovine oocyte developmental competence. Biology of Reproduction 85 6269. (https://doi.org/10.1095/biolreprod.110.088815)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • AardemaHvan TolHTAWubboltsRWBrouwersJGadellaBMRoelenBAJ 2017 Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress. Biology of Reproduction 96 982992. (https://doi.org/10.1095/biolreprod.116.146159)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • ActonBMJurisicovaAJurisicaICasperRF 2004 Alterations in mitochondrial membrane potential during preimplantation stages of mouse and human embryo development. Molecular Human Reproduction 10 2332. (https://doi.org/10.1093/molehr/gah004)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • ALJohaniAMSyedDNNtambiJM 2017 Insights into stearoyl-CoA desaturase-1 regulation of systemic metabolism. Trends in Endocrinology and Metabolism 28 831842. (https://doi.org/10.1016/j.tem.2017.10.003)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • AnderssonLBostromPEricsonJRutbergMMagnussonBMarchesanDRuizMAspLHuangPFrohmanMA et al. 2006 PLD1 and ERK2 regulate cytosolic lipid droplet formation. Journal of Cell Science 119 22462257. (https://doi.org/10.1242/jcs.02941)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • BellerMSztalrydCSouthallNBellMJackleHAuldDSOliverB 2008 COPI complex is a regulator of lipid homeostasis. PLoS Biology 6 25302549.

  • BoutetEEl MourabitHProtMNemaniMKhalloufEColardOMauriceMDurand-SchneiderAMChretienYGresS et al. 2009 Seipin deficiency alters fatty acid Delta9 desaturation and lipid droplet formation in Berardinelli-Seip congenital lipodystrophy. Biochimie 91 796803. (https://doi.org/10.1016/j.biochi.2009.01.011)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • BurnumKECornettDSPuolitaivalSMMilneSBMyersDSTranguchSBrownHADeySKCaprioliRM 2009 Spatial and temporal alterations of phospholipids determined by mass spectrometry during mouse embryo implantation. Journal of Lipid Research 50 22902298. (https://doi.org/10.1194/jlr.M900100-JLR200)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • CuiXSLiXYShenXHBaeYJKangJJKimNH 2007 Transcription profile in mouse four-cell, morula, and blastocyst: genes implicated in compaction and blastocoel formation. Molecular Reproduction and Development 74 133143. (https://doi.org/10.1002/mrd.20483)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • del ColladoMSaraivaNZLopesFLGasparRCPadilhaLCCostaRRRossiGFVantiniRGarciaJM 2016 Influence of bovine serum albumin and fetal bovine serum supplementation during in vitro maturation on lipid and mitochondrial behaviour in oocytes and lipid accumulation in bovine embryos. Reproduction Fertility and Development 28 17211732. (https://doi.org/10.1071/RD15067)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • DumollardRCampbellKHaletGCarrollJSwannK 2008 Regulation of cytosolic and mitochondrial ATP levels in mouse eggs and zygotes. Developmental Biology 316 431440. (https://doi.org/10.1016/j.ydbio.2008.02.004)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • DunningKRCashmanKRussellDLThompsonJGNormanRJRobkerRL 2010 Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biology of Reproduction 83 909918. (https://doi.org/10.1095/biolreprod.110.084145)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • FareseRVWaltherTC 2009 Lipid droplets finally get a little R-E-S-P-E-C-T. Cell 139 855860. (https://doi.org/10.1016/j.cell.2009.11.005)

  • FlowersMTNtambiJM 2008 Role of stearoyl-coenzyme A desaturase in regulating lipid metabolism. Current Opinion in Lipidology 19 248256. (https://doi.org/10.1097/MOL.0b013e3282f9b54d)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • Fouladi-NashtaAAGutierrezCGGongJGGarnsworthyPCWebbR 2007 Impact of dietary fatty acids on oocyte quality and development in lactating dairy cows. Biology of Reproduction 77 917. (https://doi.org/10.1095/biolreprod.106.058578)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • FukunishiSSujishiTTakeshitaAOhamaHTsuchimotoYAsaiATsudaYHiguchiK 2014 Lipopolysaccharides accelerate hepatic steatosis in the development of nonalcoholic fatty liver disease in Zucker rats. Journal of Clinical Biochemistry and Nutrition 54 3944. (https://doi.org/10.3164/jcbn.13-49)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • GenicotGLeroyJLMRVan SoomADonnayI 2005 The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes. Theriogenology 63 11811194. (https://doi.org/10.1016/j.theriogenology.2004.06.006)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • GuoYWaltherTCRaoMStuurmanNGoshimaGTerayamaKWongJSValeRDWalterPFareseRV 2008 Functional genomic screen reveals genes involved in lipid-droplet formation and utilization. Nature 453 657661. (https://doi.org/10.1038/nature06928)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • HaggartyPWoodMFergusonEHoadGSrikantharajahAMilneEHamiltonMBhattacharyaS 2006 Fatty acid metabolism in human preimplantation embryos. Human Reproduction 21 766773. (https://doi.org/10.1093/humrep/dei385)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • HallAMBruntEMChenZJViswakarmaNReddyJKWolinsNEFinckBN 2010 Dynamic and differential regulation of proteins that coat lipid droplets in fatty liver dystrophic mice. Journal of Lipid Research 51 554563. (https://doi.org/10.1194/jlr.M000976)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • HapalaIMarzaEFerreiraT 2011 Is fat so bad? Modulation of endoplasmic reticulum stress by lipid droplet formation. Biology of the Cell 103 271285. (https://doi.org/10.1042/BC20100144)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • HessDChisholmJWIgalRA 2010 Inhibition of stearoylCoA desaturase activity blocks cell cycle progression and induces programmed cell death in lung cancer cells. PLoS ONE 5 e11394. (https://doi.org/10.1371/journal.pone.0011394)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • HsiehKLeeYKLondosCRaakaBMDalenKTKimmelAR 2012 Perilipin family members preferentially sequester to either triacylglycerol-specific or cholesteryl-ester-specific intracellular lipid storage droplets. Journal of Cell Science 125 40674076. (https://doi.org/10.1242/jcs.104943)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • LeeDKParkCHChoiKHJeongYIUhKJHwangJYLeeSGLeeCK 2016 Aggregation of cloned embryos in empty zona pellucida improves derivation efficiency of pig ES-like cells. Zygote 24 909917. (https://doi.org/10.1017/S0967199416000241)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • LeeseHJ 2012 Metabolism of the preimplantation embryo: 40 years on. Reproduction 143 417427. (https://doi.org/10.1530/REP-11-0484)

  • LeroyJLGenicotGDonnayIVan SoomA 2005a Evaluation of the lipid content in bovine oocytes and embryos with nile red: a practical approach. Reproduction in Domestic Animals 40 7678. (https://doi.org/10.1111/j.1439-0531.2004.00556.x)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • LeroyJLMRVanholderTMateusenBChristopheAOpsomerGde KruifAVan SoomA 2005b Non-esterified fatty acids in follicular fluid of dairy cows and their effect on developmental capacity of bovine oocytes in vitro. Reproduction 130 485495. (https://doi.org/10.1530/rep.1.00735)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • LounisMABergeronKFBurhansMSNtambiJMMounierC 2017 Oleate activates SREBP-1 signaling activity in SCD1-deficient hepatocytes. American Journal of Physiology-Endocrinology and Metabolism 313 E710E720. (https://doi.org/10.1152/ajpendo.00151.2017)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • LynRKSingaraveluRKargmanSO’HaraSChanHOballaRHuangZJonesDMRidsdaleARussellRS et al. 2014 Stearoyl-CoA desaturase inhibition blocks formation of hepatitis C virus-induced specialized membranes. Scientific Reports 4 4549. (https://doi.org/10.1038/srep04549)

    • PubMed
    • Search Google Scholar
    • Export Citation
  • MarchiSPatergnaniSPintonP 2014 The endoplasmic reticulum-mitochondria connection: one touch, multiple functions. Biochimica et Biophysica Acta 1837 461469. (https://doi.org/10.1016/j.bbabio.2013.10.015)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • MinBLeeHSongJHHanMJChungJ 2014 Arctiin inhibits adipogenesis in 3T3-L1 cells and decreases adiposity and body weight in mice fed a high-fat diet. Nutrition Research and Practice 8 655661. (https://doi.org/10.4162/nrp.2014.8.6.655)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • MiyazakiMDobrzynAManWCChuKKSampathHKimHJNtambiJN 2004 Stearoyl-CoA desaturase 1 gene expression is necessary for fructose-mediated induction of lipogenic gene expression by sterol regulatory element-binding protein-1c-dependent and -independent mechanisms. Journal of Biological Chemistry 279 2516425171. (https://doi.org/10.1074/jbc.M402781200)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • MiyazakiMFlowersMTSampathHChuKOtzelbergerCLiuXNtambiJM 2007 Hepatic stearoyl-CoA desaturase-1 deficiency protects mice from carbohydrate-induced adiposity and hepatic steatosis. Cell Metabolism 6 484496. (https://doi.org/10.1016/j.cmet.2007.10.014)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • MuYMYanaseTNishiYTanakaASaitoMJinCHMukasaCOkabeTNomuraMGotoK 2001 Saturated FFAs, palmitic acid and stearic acid, induce apoptosis in human granulosa cells. Endocrinology 142 35903597. (https://doi.org/10.1210/endo.142.8.8293)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • NtambiJMMiyazakiMStoehrJPLanHKendziorskiCMYandellBSSongYCohenPFriedmanJMAttieAD 2002 Loss of stearoyl-CoA desaturase-1 function protects mice against adiposity. PNAS 99 1148211486. (https://doi.org/10.1073/pnas.132384699)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • PaiWYHsuCCLaiCYChangTZTsaiYLHerGM 2013 Cannabinoid receptor 1 promotes hepatic lipid accumulation and lipotoxicity through the induction of SREBP-1c expression in zebrafish. Transgenic Research 22 823838. (https://doi.org/10.1007/s11248-012-9685-0)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • PengGLiLLiuYPuJZhangSYuJZhaoJLiuP 2011 Oleate blocks palmitate-induced abnormal lipid distribution, endoplasmic reticulum expansion and stress, and insulin resistance in skeletal muscle. Endocrinology 152 22062218. (https://doi.org/10.1210/en.2010-1369)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • RenCFWangLZFanYXJiaRXZhangGMDengMTDengKPWangF 2018 Scd1 contributes to lipid droplets formation in GMEC via transcriptional regulation of Tip47 and Adrp. European Journal of Lipid Science and Technology 120 1700238. (https://doi.org/10.1002/ejlt.201700238)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • RocheJF 2006 The effect of nutritional management of the dairy cow on reproductive efficiency. Animal Reproduction Science 96 282296. (https://doi.org/10.1016/j.anireprosci.2006.08.007)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • ShiXLiJZouXJGreggainJRodkaerSVFaergemanNJLiangBWattsJL 2013 Regulation of lipid droplet size and phospholipid composition by stearoyl-CoA desaturase. Journal of Lipid Research 54 25042514. (https://doi.org/10.1194/jlr.M039669)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • SturmeyRGO’ToolePJLeeseHJ 2006 Fluorescence resonance energy transfer analysis of mitochondrial: lipid association in the porcine oocyte. Reproduction 132 829837. (https://doi.org/10.1530/REP-06-0073)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • SturmeyRGReisALeeseHJMcEvoyTGE 2009 Role of fatty acids in energy provision during oocyte maturation and early embryo development. Reproduction in Domestic Animals 44 (Supplement 3) 5058. (https://doi.org/10.1111/j.1439-0531.2009.01402.x)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • SudanoMJRascadoTDTataABelazKRSantosVGValenteRSMesquitaFSFerreiraCRAraujoJPEberlinMN et al. 2016 Lipidome signatures in early bovine embryo development. Theriogenology 86 472.e471484.e471.

    • Search Google Scholar
    • Export Citation
  • ThiamARFareseRVJrWaltherTC 2013 The biophysics and cell biology of lipid droplets. Nature Reviews Molecular Cell Biology 14 775786. (https://doi.org/10.1038/nrm3699)

  • WaltherTCFareseRVJr 2012 Lipid droplets and cellular lipid metabolism. Annual Review of Biochemistry 81 687714. (https://doi.org/10.1146/annurev-biochem-061009-102430)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • WilflingFWangHJHaasJTKrahmerNGouldTJUchidaAChengJXGrahamMChristianoRFrohlichFet al. 2013 Triacylglycerol synthesis enzymes mediate lipid droplet growth by relocalizing from the ER to lipid droplets. Developmental Cell 24 384399. (https://doi.org/10.1016/j.devcel.2013.01.013)

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • WolinsNEQuaynorBKSkinnerJRSchoenfishMJTzekovABickelPE 2005 S3-12, Adipophilin, and TIP47 package lipid in adipocytes. Journal of Biological Chemistry 280 1914619155. (https://doi.org/10.1074/jbc.M500978200)

    • Crossref
    • Search Google Scholar
    • Export Citation

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