Bangmin Liu, Likun Duan, Xiaojing Liu, Xiaoqiu Wang
During the
peri-implantation period of pregnancy in pigs, the rapid elongation of
conceptuses (embryonic/fetus and its extraembryonic membranes) is highly
dependent on the composition of histotroph secreted from uterine luminal (LE)
and glandular (GE) epithelial and stroma cells, as well as selective transport
of nutrients. However, little is known about the metabolites in the uterine
luminal fluid (ULF) associated with the uterine-conceptus communication during
early pregnancy in pigs. Thus, this study was conducted to profile the
metabolome in porcine ULF between days 10 and 16 of estrus cycle (C) and
pregnancy (P). Gilts were observed for estrus and/or bred via artificial
insemination at 12 and 24 h after onset of estrus (day 0). On days 10, 12, 14,
or 16 of the estrous cycle and pregnancy (n = 6 gilts per day and status),
uteri were flushed with 20 mL sterile PBS (pH 7.2) after gilts were subjected
to a midventral laparotomy. Pregnancy was confirmed by the presence of one or
more morphologically normal conceptuses. Recoverable uterine flushings (i.e.,
ULF) were then subjected to metabolomic analysis by LC-MS. Overall, 222
metabolites were detected in the ULF of which 102 were altered by status and 63
were altered by days (P < 0.05; fold change>1.5). Comparing to cyclic day
10 (10C), pregnancy stimulated increases in citric acid, lysine, arginine and
other 19 metabolites in the ULF at pregnant day 10 (10P). At 12P when porcine conceptus
initiates its morphological changes, ornithine, α-D-glucose, phenylalanine and
other 13 metabolites were up-regulated as compared with 12C. At 14P when
conceptus initiates implantation, 15 metabolites changed in ULF. At 16P, 123
metabolites were altered in ULF as compared with 16C. Interestingly, of 15
altered metabolites in ULF, 12 were downregulated including asparagine,
nicotinamide riboside, and citrulline at 14P. Whereas 102 of 123 altered
metabolites were upregulated including phosphorylcholine, 6-phosphate fructose
and 6-phosphate glucose in ULF at 16P. To determine the absolute amount of
amino acids in the ULF, we performed the targeted metabolomic analyses for 19
amino acids. As a result, arginine, asparagine, glutamic acid, glutamine,
histidine, leucine/isoleucine, lysine, phenylalanine, proline, and valine were
increased between 10P and 16P. They were less expressed in the ULF of estrus
cycle as compared with pregnancy, and remained unchanged between 10C and 16C. Pathway
analysis based on KEGG database indicating that arginine biosynthesis, alanine,
aspartate and glutamate metabolism, and aminoacyl-tRNA biosynthesis have the
highest degree of enrichment. These novel findings provide the foundation for
future investigation of metabolomic and functional studies on both endometrium
and conceptus required for elongation of porcine conceptus during
peri-implantation period of pregnancy.
2023,JAS,101:Issue Supplement_1
https://doi.org/10.1093/jas/skad068.046
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