Insulin and branched-chain amino acid depletion during mouse preimplantation embryo culture programmes body weight gain and raised blood pressure during early postnatal life

Abstract

Mouse maternal low protein diet exclusively during preimplantation development (Emb-LPD) is sufficient to programme altered growth and cardiovascular dysfunction in offspring. Here, we use an in vitro model comprising preimplantation culture in medium depleted in insulin and branched-chain amino acids (BCAA), two proposed embryo programming inductive factors from Emb-LPD studies, to examine the consequences for blastocyst organisation and, after embryo transfer (ET), postnatal disease origin. Two-cell embryos were cultured to blastocyst stage in defined KSOM medium supplemented with four combinations of insulin and BCAA concentrations. Control medium contained serum insulin and uterine luminal fluid amino acid concentrations (including BCAA) found in control mothers from the maternal diet model (N-insulin + N-bcaa). Experimental medium (three groups) contained 50% reduction in insulin and/or BCAA (L-insulin + N-bcaa, N-insulin + L-bcaa, and L-insulin + N-bcaa). Lineage-specific cell numbers of resultant blastocysts were not affected by treatment. Following ET, a combined depletion of insulin and BCAA during embryo culture induced a non sex-specific increase in birth weight and weight gain during early postnatal life. Furthermore, male offspring displayed relative hypertension and female offspring reduced heart/body weight, both characteristics of Emb-LPD offspring. Combined depletion of metabolites also resulted in a strong positive correlation between body weight and glucose metabolism that was absent in the control group. Our results support the notion that composition of preimplantation culture medium can programme development and associate with disease origin affecting postnatal growth and cardiovascular phenotypes and implicate two important nutritional mediators in the inductive mechanism. Our data also have implications for human assisted reproductive treatment (ART) practice.

Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model

“…Therefore, differences in body weight confirmed a lower growth performance in VT animals, which agreed with previous studies carried out in rabbits undergoing VET [15,30,39]. Concordantly, previous studies have demonstrated that the nature of the preimplantation environment during ART can program development, affecting postnatal growth and phenotype [7,14,[45][46][47]. Notably, although the pattern of postnatal growth alteration in VT animals was similar in both sexes, males duplicated body weight differences between VT and NC compared to the females at the age of 20 weeks.…”

High maternal folic acid intake around conception alters mouse blastocyst lineage allocation and expression of key developmental regulatory genes

“…All data analyses from the embryo studies were analyzed in parallel by a multilevel random effect regression model (Table S2) which takes into account the hierarchical nature of the data considering between‐dams and between‐litter variations and the different parameters measured from individual embryos including number of cells, gene expression, and immunofluorescence. This regression model accounting for embryo number has been widely used previously (Kwong et al, 2004; Velazquez et al, 2018; Watkins et al, 2007, 2010, 2011; Watkins, Ursell, et al, 2008; Watkins, Wilkins, et al, 2008; C. L. Williams et al, 2011). Both ANOVA (reported in tables and figures in main manuscript) and the multilevel regression model (Table S2) gave similar results.…”

“…This regression model accounting for embryo number has been widely used previously (Kwong et al, 2004;Velazquez et al, 2018;Watkins et al, 2007Watkins et al, , 2010Watkins et al, , 2011Watkins, Ursell, et al, 2008;C. L. Williams et al, 2011).…”

Periconceptional environment and the developmental origins of disease

“…Insulin and BCAAs are potent activators of the mTOR signalling pathway regulating cellular growth (Wang & Proud 2009) and, as a consequence of dietary-induced reduction in these metabolites, blastocyst mTOR activity is reduced by Emb-LPD (Eckert et al 2012). This early maternal-embryo interaction is critical since it activates later adverse programming as shown both by an in vitro culture model in medium reduced in insulin and BCAAs (Velazquez et al 2018) and by embryo transfer of Emb-LPD blastocysts into control, normal-fed, recipients (Watkins et al 2008).…”