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: -60d-7d) or preimplantation (PIUN: 0d-7d) undernutrition at 136-138d gestation (term=150d). The mRNA and protein abundance of insulin signalling 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<0.01) and protein abundance and the protein abundance of IRS-1 (P<0.01), p110β (P<0.05), PTEN (P<0.05), CREB (P<0.01) and pCREB (Ser133) (P<0.05) were decreased in the PCUN and PIUN singletons. In contrast, hepatic protein abundance of IRS-1 (P<0.01), p85 (P<0.01), p110β (P<0.001), PTEN (P<0.01), Akt2 (P<0.01), pAkt (Ser473) (P<0.01) and pFOXO-1(Thr24) (P<0.01) was increased in twins. There was a decrease in PEPCK-C mRNA (P<0.01), but paradoxically an increase in PEPCK-C protein (P<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 signalling and gluconeogenesis are explained by the changes in the expression of a suite of specific candidate microRNAs.