Abstract
Assisted reproductive technology (ART) is being increasingly applied to overcome infertility. However, the in vitro production process, the main procedure of ART, can lead to aberrant embryonic development and health-related problems in offspring. Understanding the mechanisms underlying the ART-induced side effects is important to improve the ART process. In this study, we carried out comparative transcriptome profiling between in vivo- (IVO) and in vitro- produced (IVP) mouse blastocysts. Our results suggested that aberrant actin organization might be a major factor contributing to the impaired development of IVP embryos. To test this, we examined the effect of actin disorganization on the development of IVP preimplantation embryos. Specific disruption of actin organization by cytochalasin B (CB) indicated that well-organized actin is essential for in vitro embryonic development. Supplementing the culture medium with 10–9 M melatonin, a cytoskeletal modulator in adult somatic cells, significantly reversed the disrupted expression patterns of genes related to actin organization, including Arhgef2, Bcl2, Coro2b, Flnc, and Palld. Immunofluorescence analysis showed that melatonin treatment of IVP embryos significantly improved the distribution and organization of actin filaments (F-actin) from the 8-cell stage onwards. More importantly, we found that melatonin alleviated the CB-mediated aberrant F-actin distribution and organization and rescued CB-induced impaired embryonic development. This is the first study to indicate that actin disorganization is implicated in impaired development of IVP embryos during the preimplantation stage. We also demonstrated that improving actin organization is a promising strategy to optimize existing IVP systems.
Highlights
The number of patients receiving assisted reproductive technology (ART) to treat infertility has increased annually
Clusters involved in actin cytoskeleton organization and the regulation of actin filament-based processes were located at the center of the network
These results indicated that cytoskeletal organization, especially actin organization, was disrupted in in vitro- produced (IVP) blastocysts
Summary
The number of patients receiving assisted reproductive technology (ART) to treat infertility has increased annually. Using high-throughput methods, such as microarrays and RNA sequencing (RNA-seq), many studies have reported that a series of biological processes, including energy metabolism [5], genetic information processing [7], and epigenetic modifications [8, 9], are disrupted in IVP embryos. These deductions were generally based on the functional analysis of high-throughput data, which need to be further confirmed. In-depth understanding of mechanisms of these effects would be beneficial to optimize the IVP processes
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