Protective Effect of Minocycline Hydrochloride on the Mouse Embryonic Development Against Suboptimal Environment.

Xiaojing Hou,Yang Yang,Changming Cai,Shiyu An,Shuqin Zhao,Hao Sun,Yuanlin He

doi:10.3389/fcell.2022.799042

Abstract

Numerous studies have reported how inner cell mass (ICM) and trophectoderm (TE) was determined during the process of early mouse embryonic development from zygotes into organized blastocysts, however, multiple mysteries still remain. It is noteworthy that pluripotent stem cells (PSCs), which are derived from embryos at different developmental stages, have identical developmental potential and molecular characteristics to their counterpart embryos. Advances of PSCs research may provide us a distinctive perspective of deciphering embryonic development mechanism. Minocycline hydrochloride (MiH), a critical component for maintaining medium of novel type of extended pluripotent stem cells, which possesses developmental potential similar to both ICM and TE, can be substituted with genetic disruption of Parp1 in our previous study. Though Parp1-deficient mouse ESCs are more susceptible to differentiate into trophoblast derivatives, what role of MiH plays in mouse preimplantation embryonic development is still a subject of concern. Here, by incubating mouse zygotes in a medium containing MiH till 100 h after fertilization, we found that MiH could slow down embryonic developmental kinetics during cleavage stage without impairing blastocyst formation potential. Olaparib and Talazoparib, two FDA approved PARP1 inhibitors, exhibited similar effects on mouse embryos, indicating the aforementioned effects of MiH were through inhibiting of PARP1. Besides, we showed an embryonic protective role of MiH against suboptimal environment including long term exposure to external environment and H2O2 treatment, which could mimic inevitable manipulation during embryo culture procedures in clinical IVF laboratory. To our knowledge, it is not only for the first time to study MiH in the field of embryo development, but also for the first time to propose MiH as a protective supplement for embryo culture, giving the way to more studies on exploring the multiple molecular mechanisms on embryonic development that might be useful in assisted reproductive technology.

Highlights

During early murine embryo development, the zygote undergoes a serials of cell divisions and generates a blastocyst which consists of two distinct cell types: the inner cell mass (ICM) and the trophectoderm (TE) that surrounds the ICM (Johnson and Ziomek 1981; Fleming 1987; Morris et al, 2010)
Parp1-deficient mouse extended pluripotent stem cells (EPS) cultured in condition without minocycline hydrochloride (MiH) could develop into both TE and ICM of the embryos as well (Yang et al, 2017), indicating MiH performed through inhibiting Poly (ADP-ribose) polymerase-1 (PARP1), which was consistent with the work of Alano et al, 2006
To decipher the effect of MiH on preimplantation mouse embryos, in vitro fertilization (IVF) was employed to obtain zygotes, which were randomized into two groups and cultured with KSOM medium in the absence or presence of 2 μM MiH (MiH group) till most of them developed into blastocysts at 100 h after fertilization (Figure 1A)

Summary

Introduction

During early murine embryo development, the zygote undergoes a serials of cell divisions and generates a blastocyst which consists of two distinct cell types: the inner cell mass (ICM) and the trophectoderm (TE) that surrounds the ICM (Johnson and Ziomek 1981; Fleming 1987; Morris et al, 2010). In our previous study, extended pluripotent stem cells (EPS) were derived from blastocyst and exhibited widespread bi-potency for both embryonic and extraembryonic lineages in vivo. This novel type of pluripotent stem cells (PSCs) was maintained with a chemical cocktail, consisting of human leukemia inhibitory factor (hLIF), CHIR 99021, (S)-(+)-dimethindene maleate (DiM) and minocycline hydrochloride (MiH), which was short for LCDM (Yang et al, 2017). Parp1-deficient mouse EPS cultured in condition without MiH could develop into both TE and ICM of the embryos as well (Yang et al, 2017), indicating MiH performed through inhibiting Poly (ADP-ribose) polymerase-1 (PARP1), which was consistent with the work of Alano et al, 2006. What role MiH plays in the embryonic development procedure and whether it works through PARP1 inhibition are subjects of concern

Methods

Results

Conclusion