Alterations in the uterine echotexture, hemodynamics, and histological findings in relation to metabolomic profiles in goats with different ovarian activities (active versus inactive ovaries).

Abstract

This study investigated, for the first time, the alterations in the uterine echotexture and blood flow in cyclic and acyclic (inactive ovary) goats using ultrasonography. The study aimed also to evaluate the metabolomic changes in the plasma of cyclic and acyclic goats. Furthermore, the histopathological approach was applied to the specimens of the uterus to validate the findings of this study. Based on monitoring the estrous cyclicity, goats were assigned into either a cyclic group or an acyclic one (n = 7, each). Ovarian morphometry and hemodynamics were assessed to confirm group assignment. Full ultrasonographic examinations were performed to assess the uterine echotexture by B-mode ultrasonography and uterine hemodynamics by color Doppler ultrasonography in the cyclic group (at days 10-12) and acyclic group. Additionally, blood samples were withdrawn for measuring hormonal concentrations and for metabolomics analysis. Specimens of the uterus were executed for histopathological evaluation in both groups. Results revealed alterations in the uterine hemodynamics and endometrial echotexture. Goats in the cyclic group attained a significantly higher color pixel area of the endometrium compared to those in the acyclic one (P< 0.001). However, the pixel intensity of the endometrium echotexture was significantly (P< 0.05) lower in the cyclic group than in the smooth inactive ovary one. There were significant (P< 0.05) increases in the concentrations of FSH, LH, and inhibin in the cyclic group compared to their concentrations in the acyclic one. Goats in the acyclic group attained noticeable (P< 0.001) lower concentrations of E₂ and P₄ than in the cyclic goats. The metabolomic results revealed the existence of several up- and down-regulated metabolites among the studied groups. In this investigation, untargeted metabolomic analysis revealed the existence of 5 up-regulated metabolites (ketoleucine, L-fucose, D-glucurono-6,3-lactone, melatonin, and 5-methoxy tryptamine) and 5 down-regulated ones (p-octopamine, 3-hydroxyisovaleric acid, methylmalonic acid, 4-hydroxyphenylpyruvic acid, and cadaverine) in the cyclic group compared to the acyclic one. The enrichment analysis of the significant metabolites showed top pathways that may be involved in these changes, such as fructose and mannose metabolism, valine. Leucine, and isoleucine biosynthesis, linoleic acid metabolism, arginine biosynthesis, and vitamin B6 metabolism based on the KEGG enriched pathway. Altogether, the histopathological assessment showed noticeable changes in the columnar epithelial lining of the endometrial epithelium, endometrial vascularity, and endometrial glands among the studied groups. In conclusion, this study extrapolated the differences between cyclic goats (during the mid-luteal phase) and acyclic ones in terms of hormonal, hemodynamics, echotexture of the uterus, and circulating metabolomics. These findings are very crucial to fully assess the fertility potential in goats.