Abnormal Glucose Metabolism in Male Mice Offspring Conceived by in vitro Fertilization and Frozen-Thawed Embryo Transfer

Abstract

Frozen and thawed embryo transfer (FET) is currently widely applied in routine assisted reproductive technology (ART) procedure. It is of great necessity to assess the safety of FET and investigate the long-term effect including glucose metabolism on FET-conceived offspring. The mouse model is a highly efficient method to figure out the relationship between the process of FET and offspring health. In this study, we obtained mouse offspring of natural conception (NC), in vitro fertilization (IVF), and FET. Glucose and insulin tolerance test (GTT/ITT) were performed on both chow fed or high fat diet (HFD) fed offspring to examine the glucose metabolism status. We detected hepatic PI3K/AKT pathway by western blotting and transcriptome status by RNA-sequencing. Impaired glucose tolerance (IGT) and decreased insulin tolerance were occurred in FET conceived male offspring. After challenged with the HFD-fed, male offspring in FET group performed earlier and severer IGT than IVF group. Furthermore, higher HOMA-IR index and higher serum insulin level post glucose injected in FET-chow group suggested the insulin resistance status. The PI3K/AKT signaling pathway, the major pathway of insulin in the liver, were also disrupted in FET group. Transcriptomics of the liver reveals significantly downregulated in glucose metabolic process and insulin resistance in the FET-chow group. In our study, FET-conceived male mouse offspring presented glucose metabolism dysfunction mainly manifesting insulin resistance. The hepatic insulin signaling pathway were in concordance with reduced glycogen synthesis, increased glycolysis and enhanced gluconeogenesis status in FET-conceived male offspring.

Early Embryo Exposure to Assisted Reproductive Manipulation Induced Subtle Changes in Liver Epigenetics with No Apparent Negative Health Consequences in Rabbit

“…In agreement with our findings, several studies described non-polar and semi-polar metabolism alteration after embryonic manipulation [ 10 , 20 , 21 , 22 , 23 , 24 , 25 ]. Globally, semi-polar metabolites were up-accumulated while non-polar metabolites were down-accumulated in MOVET animals compared to NC animals, suggesting a metabolic adaptation strategy [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. Related to this, Feuer et al observed that conception by IVF reduces growth, impairs glucose tolerance and impacts the serum and liver metabolome, affecting a wide range of compounds such as carbohydrates, amino acids, and, especially, lipids [ 10 , 23 , 24 ].…”

“…Investigating the effects of ART in fertile model species circumvents issues raised by underlying infertility and avoided other confounding factors, especially those regarding birth weight and growth trajectories, metabolic health, cardiovascular troubles and epigenetic alterations [ 15 ]. The mechanism underlying the alterations in offspring conceived with ARTs are not entirely clear, but epigenetic changes are a well-excepted contributor [ 2 , 5 , 14 , 15 , 16 , 17 ]. Here, we exploit our rabbit ARTs model to conduct a hepatic DNA methylation association study with metabolomics and proteomics in the F3 generation for embryonic exposure (multiple ovulation, vitrification and embryo transfer) conceived offspring, as well as assess the potential health implications.…”

Anxiety and depression-like behaviours are more frequent in aged male mice conceived by ART compared with natural conception

“…Two-cell embryos in IVF-ET and IVF-FET groups were obtained by standard B6D2F1 × B6D2F1 IVF. The IVF, embryo transfer, embryo frozen, and thawed procedures have been described previously ( Qin et al 2021 ). The offspring were weaned and separated by sex until postnatal day 20.…”

Cryopreservation of Gametes and Embryos and Their Molecular Changes

“…These changes are characterized by insulin resistance and alterations of the function of proteins related to glucose metabolism. Therefore, these changes could be a risk factor for metabolic diseases as type 2 diabetes [ 66 , 67 , 68 ].…”