Summary of Study ST003120
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001938. The data can be accessed directly via it's Project DOI: 10.21228/M86T69 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST003120 |
| Study Title | Mannose is crucial for mesoderm specification and symmetry breaking in gastruloids. |
| Study Summary | Patterning and growth are fundamental features of embryonic development that must be tightly coordinated. To understand how metabolism impacts early mesoderm development, we used mouse embryonic stem cell-derived gastruloids, that co-expressed glucose transporters with the mesodermal marker T/Bra. While the glucose mimic, 2-deoxy-D-glucose (2-DG), blocked T/Bra expression and abolished axial elongation in gastruloids, removal of glucose did not phenocopy 2-DG treatment despite a decline in glycolytic intermediates occurring under both conditions. As 2-DG could also act as a competitive inhibitor of mannose in protein glycosylation, we added mannose together with 2-DG and found that it could rescue the mesoderm specification both in vivo and in vitro. We further showed that blocking production and intracellular recycling of mannose abrogated mesoderm specification. Proteomics analysis revealed that mannose reversed glycosylation of the Wnt pathway regulator, Secreted Frizzled Receptor, Frzb. Our study showed how mannose is crucial for mesoderm specification in gastruloids. |
| Institute | Dept of Genetics, University of Cambridge |
| Last Name | Dingare |
| First Name | Chaitanya |
| Address | Downing Site, Cambridge, Cambridgeshire, CB2 3EH, United Kingdom |
| cd705@cam.ac.uk | |
| Phone | +447916677460 |
| Submit Date | 2024-02-24 |
| Publications | https://doi.org/10.1101/2023.06.05.543730 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-03-13 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN005114 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Vanquish UHPLC |
| Column | Atlantis Premier BEH Z-HILIC (100 x 2.1 mm, 1.7 um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Fisher Scientific Orbitrap Exploris 240 |
| Ion Mode | NEGATIVE |
| Units | counts per second (cps) |
Chromatography:
| Chromatography ID: | CH003870 |
| Chromatography Summary: | Snap-frozen gastruloids were sent to EMBL-Heidelberg, Germany, for the metabolomics analysis. Reagents: LC-MS grade water, acetonitrile and methanol were obtained from Th. Geyer (Germany). High-purity ammonium acetate, ammonium hydroxide, and formic acid were purchased from Merck (Germany). Stable isotope labelled amino acids (MSK-MET1-1; Cambridge Isotope Laboratories, MA, USA) were used as internal standards for untargeted metabolomics. LC-MS/MS analysis was performed on a Vanquish UHPLC system coupled to an Orbitrap Exploris 240 high-resolution mass spectrometer (Thermo Fisher Scientific, MA, USA) in negative ESI (electrospray ionization) mode. Chromatographic separation was carried out on an Atlantis Premier BEH Z-HILIC column (Waters, MA, USA; 2.1?mm x 100 mm, 1.7 µm) at a flow rate of 0.25 mL/min. The mobile phase consisted of water:acetonitrile (9:1, v/v; mobile phase A) and acetonitrile:water (9:1, v/v; mobile phase B), which were modified with a total buffer concentration of 10 mM ammonium acetate. The aqueous portion of each mobile phase was adjusted to pH 9.0 via addition of ammonium hydroxide. The following gradient (20 min total run time including re-equilibration) was applied (time[min]/%B): 0/95, 2/95, 14.5/60, 16/60, 16.5/95, 20/95. Column temperature was maintained at 40°C, the autosampler was set to 4°C and sample injection volume was 7 µL. |
| Methods Filename: | LC_MS_MS_Full_Protocol.pdf |
| Instrument Name: | Vanquish UHPLC |
| Column Name: | Atlantis Premier BEH Z-HILIC (100 x 2.1 mm, 1.7 um) |
| Column Temperature: | 40 |
| Flow Gradient: | time [min]/%B - 0/95, 2/95, 14.5/60, 16/60, 16.5/95, 20/95 |
| Flow Rate: | 0.25mL/min |
| Solvent A: | water:acetonitrile 9:1, v/v; 10mM ammonium acetate |
| Solvent B: | acetonitrile:water 9:1, v/v; 10mM ammonium acetate |
| Chromatography Type: | HILIC |
