GDF-8 improves in vitro implantation and cryo-tolerance by stimulating the ALK5-SMAD2/3 signaling in bovine IVF embryo development

Seon-Min Kang; Muhammad Idrees; Chalani Dilshani Perera; Seo-Hyun Lee; Mingjun Zhang; Xianfeng Yu; Yongxun Jin; Il-Keun Kong

doi:10.3389/fcell.2024.1345669

GDF-8 improves in vitro implantation and cryo-tolerance by stimulating the ALK5-SMAD2/3 signaling in bovine IVF embryo development

Front Cell Dev Biol. 2024 Mar 21:12:1345669. doi: 10.3389/fcell.2024.1345669. eCollection 2024.

Authors

Seon-Min Kang¹, Muhammad Idrees^{1

2}, Chalani Dilshani Perera¹, Seo-Hyun Lee¹, Mingjun Zhang³, Xianfeng Yu³, Yongxun Jin³, Il-Keun Kong^{1

2}

Affiliations

¹ Division of Applied Life Science (BK21 Four), Graduate School of Applied Life Science, Gyeongsang National University, Jinju, Republic of Korea.
² Division of Animal Science, Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, Republic of Korea.
³ Jilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun, China.

Abstract

Transforming growth factor-beta (TGF-β) plays a critical role in regulating trophoblast invasion and proliferation. Growth differentiation factor-8 (GDF-8) is a member of the TGF-β superfamily and is categorized as a myostatin subtype. It is primarily a secreted protein synthesized in skeletal muscle cells. It is expressed in the placenta, reproductive tissues, and cells. In this study, we investigated the role of GDF-8 in the development and hatching rate of bovine embryos. We noted a notable elevation (p < 0.05) in the development and hatching rates compared to the control embryos. Furthermore, the GDF-8 group showed a significantly improved total cell number (p < 0.05) and an increase in trophectoderm ratio inner cell mass (trophectoderm: inner cell mass) cells (p < 0.001) compared to the control group. Additionally, blastocysts treated with GDF-8 exhibited significantly higher mRNA levels of caudal-type homeobox 2 (CDX2) (p < 0.05). The trophoblast invasion area was significantly larger in the GDF-8 group than in the control group (p < 0.01). Furthermore, qRT-PCR analysis revealed significantly higher mRNA levels (p < 0.05) of matrix metalloproteinases 9 (MMP9) and follistatin-like 3(FSTL3), both of which are associated with the ALK5-SMAD2/3 signaling pathway, in the GDF-8 group than those in the control group. The mRNA expression levels of genes related to tight junctions (TJ) and adherent junctions were higher in the GDF-8 group than those in the control group (p < 0.05). After 24 h of thawing, blastocysts were analyzed using 4-kDa FITC-dextran, which revealed a higher TJ integrity in the GDF-8 group (p < 0.01). Thus, GDF-8 plays a crucial role in bovine embryonic development, in vitro implantation, and cryotolerance.

Keywords: ALK5-SMAD2/3 signaling; GDF-8; bovine; cryotolerance; embryo development; in vitro implantation.

Grants and funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was partially supported by a National Research Foundation of Korea (NRF) grant funded by the Korean Government (MSIT; no. RS-2023-00208894), Cooperative Research Program for Agriculture Science and Technology Development (Project title: Carbon Reduction Korean Beef Selection and Carbon Emission Evaluation Base Construction Research, Project No. RS-2023-00237137)", Rural Development Administration, Republic of Korea, and a scholarship from BK21. Institutional Review Board Statement: The experiments were carried out under the guidelines of GNU and the Institutional Animal Care and Use Committee (Approval ID: GNU-230425-A0088).