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

Miguel A Velazquez,Bhavwanti Sheth,Stephanie J Smith,Judith J Eckert,Clive Osmond,Tom P Fleming

doi:10.1016/j.bbadis.2017.11.020

Miguel A Velazquez, Bhavwanti Sheth + Show 4 more

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https://doi.org/10.1016/j.bbadis.2017.11.020

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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.

Highlights

Undernutrition is a worldwide concern affecting countries with developing and emerging economies and populations in countries with a high human development index [1]
The main objective of our study was to determine whether insulin and/or branched-chain amino acids (BCAA) depletion during early development of the embryo can act as inductive factors of altered phenotypes during postnatal life
The current in vitro embryo culture (IVEC)-embryo transfer (ET) model, using the same MF1 mouse strain as used in the Emb-LPD model, attempted in the control group (N-insulin + N-bcaa) to mimic the embryo compositional environment for both amino acids (AA) detected in uterine luminal fluid (ULF) and the physiological level of serum insulin found in control diet mothers

Summary

Introduction

Undernutrition is a worldwide concern affecting countries with developing and emerging economies and populations in countries with a high human development index [1]. In a murine model of protein restriction it was shown that dams fed with a low protein diet (9% casein) exclusively during the preimplantation period (days 0–3.5 of embryonic development; Emb-LPD) produced offspring that displayed altered phenotypes during postnatal life, including increased postnatal growth, high blood pressure, vascular dysfunction and hyperactive behavior [5,6,7]. This animal model of undernutrition has revealed that compensatory mechanisms exist to maintain viable growth of the developing fetus by altering cellular characteristics of the placental lineages. These phenotypic alterations seem to be induced at the blastocyst stage, around the time of cell lineage determination, since recipients fed with normal levels of protein receiving protein-restricted embryos through embryo transfer produced conceptuses with increased weight [5]

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