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Immunohistochemistry of Leukemia Inhibitory Factor and Integrin αVβ3 in Mouse Endometrium Following Kisspeptin-54 Ovulation Trigger

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Abstract

Kisspeptin (KP) is a group of hypothalamic neuropeptides encoded by KISS-1 gene. KP-54, a 54-amino-acid peptide, helps regulate the hypothalamic-pituitary-ovarian axis and plays a potential role in implantation. C57BL/6 J female mice were superovulated via intraperitoneal injection of 5 International Units (IU) pregnant mare serum gonadotrophin (day 1). Forty-eight hours later, mice (5/group) were injected with phosphate-buffered saline (PBS) (group A), 5 IU human chorionic gonadotrophin (hCG) (group B), or 3 nmol KP-54 (group C). On day 7, mice were euthanized and uteri excised to create paraformaldehyde-fixed paraffin-embedded sections that were immunostained for the implantation markers: leukemia inhibitory factor (LIF) and integrin αVβ3 (ITG αVβ3). Slides were scored for intensity of staining in endometrial glandular epithelium (GE) and stromal cells (SCs) via histoscore (H-score). Data were analyzed using the Kruskal–Wallis test followed by the Mann–Whitney U test for pairwise comparisons. LIF expression was significantly higher in GE and SCs of mice triggered with KP-54 compared to placebo (P = .009 for both), but only higher than hCG trigger group in SCs (P = .009). Meanwhile, ITG αVβ3 expression was significantly higher in SCs of mice triggered with KP-54 compared to placebo (P = .028). In conclusion, using KP-54 as an ovulation trigger resulted in higher expression of the implantation markers LIF and ITG αVβ3 in mice endometrium compared to hCG or placebo. This suggests a potential role for KP-54 trigger in improving embryo implantation in clinical IVF. However, further studies are needed to correlate these results with clinical implantation rates and pregnancy outcomes.

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Data Availability

The data that support the findings of this study are available from the corresponding author.

The research ethics board at the University of British Columbia approved laboratory animal care and use (#A16-0295).

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Funding

The Department of Obstetrics & Gynaecology at the University of British Columbia financially supported this study.

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Authors and Affiliations

  1. Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynaecology, Faculty of Medicine, University of British Columbia, Vancouver, BC, Canada

    Amr O. Abdelkareem, Mahmoud S. Iews, Ruth Habte & Mohamed A. Bedaiwy

  2. Department of Obstetrics and Gynecology, Faculty of Medicine, Sohag University, Sohag, Egypt

    Amr O. Abdelkareem, Abdou S. Ait-Allah, Salah M. Rasheed & Yasser A. Helmy

  3. Department of Obstetrics and Gynecology, Faculty of Medicine, South Valley University, Qena, Egypt

    Mahmoud S. Iews

  4. Department of Obstetrics and Gynecology, Faculty of Medicine, Assiut University, Assiut, Egypt

    Faten F. Abdelhafez

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  1. Amr O. Abdelkareem
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  2. Mahmoud S. Iews
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Contributions

A.S. Ait-Allah and M.A. Bedaiwy were involved in idea conceptualization. A.O. Abdelkareem, A.S. Ait-Allah, S.M. Rasheed, Y.A. Helmy, and M.A. Bedaiwy were involved in study design. A.O. Abdelkareem was involved in animal experiments, immunohistochemistry, and data analysis. M.S. Iews was involved in animal experiments. A.O. Abdelkareem and R. Habte were involved in manuscript writing. R. Habte, F.F. Abdelhafez, and M.A. Bedaiwy were involved in manuscript critical revision and submission.

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Correspondence to Mohamed A. Bedaiwy.

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Conflict of Interest

Dr. Bedaiwy has received grants from the Canadian Institutes of Health Research & Ferring Pharmaceuticals. He is also on the advisory boards of AbbVie and Baxter. All other authors have no competing interests to declare.

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Abdelkareem, A.O., Iews, M.S., Ait-Allah, A.S. et al. Immunohistochemistry of Leukemia Inhibitory Factor and Integrin αVβ3 in Mouse Endometrium Following Kisspeptin-54 Ovulation Trigger. Reprod. Sci. 30, 3084–3091 (2023). https://doi.org/10.1007/s43032-023-01243-7

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