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

in Reproduction
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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.


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