在经历形态变化的 3D 藻酸盐水凝胶培养系统中包封的猪囊胚分泌代谢组提供了对猪胚胎延伸起始过程中涉及的特定机制的深入了解。

Secreted metabolome of porcine blastocysts encapsulated within 3D alginate hydrogel culture systems undergoing morphological changes provides insights into specific mechanisms involved in the initiation of porcine conceptus elongation.

机构信息

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.

出版信息

Reprod Fertil Dev. 2023 Mar;35(5):375-394. doi: 10.1071/RD22210.

DOI:10.1071/RD22210

PMID:36780705

Abstract

CONTEXT

The exact mechanisms regulating the initiation of porcine conceptus elongation are not known due to the complexity of the uterine environment.

AIMS

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.

METHODS

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).

KEY RESULTS

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.

CONCLUSIONS

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.

IMPLICATIONS

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.

摘要

背景

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。

目的

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。

方法

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。