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

Reprod Sci. 2023 Oct;30(10):3084-3091. doi: 10.1007/s43032-023-01243-7. Epub 2023 May 1.

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.

Keywords: Immunohistochemistry; Implantation; Integrin αVβ3; Kisspeptin; LIF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chorionic Gonadotropin / metabolism
  • Chorionic Gonadotropin / pharmacology
  • Embryo Implantation / physiology
  • Endometrium / metabolism
  • Female
  • Horses
  • Humans
  • Immunohistochemistry
  • Integrin alphaVbeta3* / metabolism
  • Kisspeptins* / metabolism
  • Leukemia Inhibitory Factor / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Ovulation
  • Ovulation Induction / methods
  • Pregnancy

Substances

  • Integrin alphaVbeta3
  • Kisspeptins
  • Leukemia Inhibitory Factor
  • Chorionic Gonadotropin