Long-term and transgenerational phenotypic, transcriptional and metabolic effects in rabbit males born following vitrified embryo transfer

Francisco Marco-Jiménez,Joaquín Cañizares,José S. Vicente,David S. Peñaranda,Gianfranco Diretto,Ximo Garcia-Dominguez,Víctor García-Carpintero

doi:10.1038/s41598-020-68195-9

Abstract

The advent of assisted reproductive technologies (ART) in mammals involved an extraordinary change in the environment where the beginning of a new organism takes place. Under in vitro conditions, in which ART is currently being performed, it likely fails to mimic optimal in vivo conditions. This suboptimal environment could mediate in the natural developmental trajectory of the embryo, inducing lasting effects until later life stages that may be inherited by subsequent generations (transgenerational effects). Therefore, we evaluated the potential transgenerational effects of embryo exposure to the cryopreservation-transfer procedure in a rabbit model on the offspring phenotype, molecular physiology of the liver (transcriptome and metabolome) and reproductive performance during three generations (F1, F2 and F3). The results showed that, compared to naturally-conceived animals (NC group), progeny generated after embryo exposure to the cryopreservation-transfer procedure (VT group) exhibited lower body growth, which incurred lower adult body weight in the F1 (direct effects), F2 (intergenerational effects) and F3 (transgenerational effects) generations. Furthermore, VT animals showed intergenerational effects on heart weight and transgenerational effects on liver weight. The RNA-seq data of liver tissue revealed 642 differentially expressed transcripts (DETs) in VT animals from the F1 generation. Of those, 133 were inherited from the F2 and 120 from the F3 generation. Accordingly, 151, 190 and 159 differentially accumulated metabolites (DAMs) were detected from the F1, F2 and F3, respectively. Moreover, targeted metabolomics analysis demonstrated that transgenerational effects were mostly presented in the non-polar fraction. Functional analysis of molecular data suggests weakened zinc and fatty acid metabolism across the generations, associated with alterations in a complex molecular network affecting global hepatic metabolism that could be associated with the phenotype of VT animals. However, these VT animals showed proper reproductive performance, which verified a functional health status. In conclusion, our results establish the long-term transgenerational effects following a vitrified embryo transfer procedure. We showed that the VT phenotype could be the result of the manifestation of embryonic developmental plasticity in response to the stressful conditions during ART procedures.

Highlights

The advent of assisted reproductive technologies (ART) in mammals involved an extraordinary change in the environment where the beginning of a new organism takes place
No previous reports have examined the transgenerational effects of the embryo cryopreservation-transfer procedure (CTP)
For the first time, the transgenerational inheritance of changes induced by embryo exposure to CTP over the offspring growth performance, adult body weight, phenotype of vital organs and their molecular physiology and metabolism

Summary

Introduction

The advent of assisted reproductive technologies (ART) in mammals involved an extraordinary change in the environment where the beginning of a new organism takes place. In which ART is currently being performed, it likely fails to mimic optimal in vivo conditions This suboptimal environment could mediate in the natural developmental trajectory of the embryo, inducing lasting effects until later life stages that may be inherited by subsequent generations (transgenerational effects). It is well accepted that in responding to environmental cues the embryo demonstrates a high degree of developmental plasticity, modulating its metabolism, gene expression and p henotype[4] This embryo adaptation is thought to increase short-term survival in suboptimal environments during ART, several studies have associated this embryo developmental deviation with adverse consequences later in life (Developmental Origins of Health and Disease theory), as has been extensively described in h umans[5,6,7,8,9,10,11]. Taking all these data into account, the aim in this work is to determine whether embryo exposure to a CTP can alter offspring features and molecular signatures across three generations (transgenerational effects)

Methods

Results

Conclusion