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This study aimed to determine whether exposure of the oocyte and/or embryo to maternal undernutrition results in the later programming of insulin action in the liver and factors regulating gluconeogenesis. To do this, we collect livers from singleton and twin fetal sheep that were exposed to periconceptional (PCUN; -60 to 7 days) or preimplantation (PIUN; 0-7 days) undernutrition at 136-138 days of gestation (term = 150 days). The mRNA and protein abundance of insulin signaling and gluconeogenic factors were then quantified using qRT-PCR and Western blotting, respectively, and global microRNA expression was quantified using deep sequencing methodology. We found that hepatic PEPCK-C mRNA (P &lt; 0.01) and protein abundance and the protein abundance of IRS-1 (P &lt; 0.01), p110β (P &lt; 0.05), PTEN (P &lt; 0.05), CREB (P &lt; 0.01), and pCREB (Ser(133); P &lt; 0.05) were decreased in the PCUN and PIUN singletons. In contrast, hepatic protein abundance of IRS-1 (P &lt; 0.01), p85 (P &lt; 0.01), p110β (P &lt; 0.001), PTEN (P &lt; 0.01), Akt2 (P &lt; 0.01), p-Akt (Ser(473); P &lt; 0.01), and p-FOXO-1 (Thr24) (P &lt; 0.01) was increased in twins. There was a decrease in PEPCK-C mRNA (P &lt; 0.01) but, paradoxically, an increase in PEPCK-C protein (P &lt; 0.001) in twins. Both PCUN and PIUN altered the hepatic expression of 23 specific microRNAs. We propose that the differential impact of maternal undernutrition in the presence of one or two embryos on mRNAs and proteins involved in the insulin signaling and gluconeogenesis is explained by changes in the expression of a suite of specific candidate microRNAs.

endocrinology and metabolism/american journal of physiology: endocrinology and metabolism

Impact of embryo number and maternal undernutrition around the time of conception on insulin signaling and gluconeogenic factors and microRNAs in the liver of fetal sheep