Department of Biological Systems Engineering, University of Nebraska-Lincoln, P.O. Box 830726, Lincoln, NE 68583, USA.
USDA, U.S. Meat Animal Research Center, P.O. Box 166, Clay Center, NE 68933, USA.
Reprod Fertil Dev. 2023 Mar;35(5):375-394. doi: 10.1071/RD22210.
The exact mechanisms regulating the initiation of porcine conceptus elongation are not known due to the complexity of the uterine environment.
To identify contributing factors for initiation of conceptus elongation in vitro , this study evaluated differential metabolite abundance within media following culture of blastocysts within unmodified alginate (ALG) or Arg-Gly-Asp (RGD)-modified alginate hydrogel culture systems.
Blastocysts were harvested from pregnant gilts, encapsulated within ALG or RGD or as non-encapsulated control blastocysts (CONT), and cultured. At the termination of 96h culture, media were separated into blastocyst media groups: non-encapsulated control blastocysts (CONT); ALG and RGD blastocysts with no morphological change (ALG- and RGD-); ALG and RGD blastocysts with morphological changes (ALG+ and RGD+) and evaluated for non-targeted metabolomic profiling by liquid chromatography (LC)-mass spectrometry (MS) techniques and gas chromatography-(GC-MS).
Analysis of variance identified 280 (LC-MS) and 1 (GC-MS) compounds that differed (P <0.05), of which 134 (LC-MS) and 1 (GC-MS) were annotated. Metabolites abundance between ALG+ vs ALG-, RGD+ vs RGD-, and RGD+ vs ALG+ were further investigated to identify potential differences in metabolic processes during the initiation of elongation.
This study identified changes in phospholipid, glycosphingolipid, lipid signalling, and amino acid metabolic processes as potential RGD-independent mechanisms of elongation and identified changes in lysophosphatidylcholine and sphingolipid secretions during RGD-mediated elongation.
These results illustrate changes in phospholipid and sphingolipid metabolic processes and secretions may act as mediators of the RGD-integrin adhesion that promotes porcine conceptus elongation.
由于子宫环境的复杂性，调控猪胚胎延伸起始的确切机制尚不清楚。
为了鉴定体外胚胎延伸起始的影响因素，本研究评估了在未经修饰的藻酸盐（ALG）或 Arg-Gly-Asp（RGD）修饰的藻酸盐水凝胶培养系统中培养囊胚后培养基中差异代谢物丰度。
从怀孕母猪中收获囊胚，将其包裹在 ALG 或 RGD 中或作为未包裹的对照囊胚（CONT）进行培养。在 96h 培养结束时，将培养基分为囊胚培养基组：未包裹的对照囊胚（CONT）；无形态变化的 ALG 和 RGD 囊胚（ALG-和 RGD-）；形态变化的 ALG 和 RGD 囊胚（ALG+和 RGD+），并通过液相色谱（LC）-质谱（MS）技术和气相色谱-（GC-MS）评估非靶向代谢组学分析。
方差分析鉴定出 280 种（LC-MS）和 1 种（GC-MS）差异（P <0.05）的化合物，其中 134 种（LC-MS）和 1 种（GC-MS）进行了注释。进一步研究了 ALG+与 ALG-、RGD+与 RGD-和 RGD+与 ALG+之间的代谢物丰度，以鉴定伸长起始时代谢过程中的潜在差异。
本研究鉴定出磷脂、糖脂、脂质信号和氨基酸代谢过程的变化可能是伸长的 RGD 独立机制，并且在 RGD 介导的伸长过程中鉴定出溶血磷脂酰胆碱和鞘脂的分泌变化。
这些结果表明，磷脂和鞘脂代谢过程和分泌的变化可能作为促进猪胚胎伸长的 RGD-整合素黏附的介质。
