Summary of Study ST002507
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 PR001619. The data can be accessed directly via it's Project DOI: 10.21228/M8F41P 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 | ST002507 |
| Study Title | Time-course analysis of C13 labeling in mouse eye organoids. |
| Study Summary | The direct quantification of glucose consumption using 13C glucose time-course tracing was performed in cultured eye organoids and measured by LC-MS/MS analysis. The incubation was performed from 15 minutes to 2 hours. Cell Name AES0145 : Rx-GFP K/I EB5 (RIKEN Cell Bank): Organoid method (PMID: 21475194). |
| Institute | Northwestern University |
| Last Name | TAKATA |
| First Name | NOZOMU |
| Address | 303 East Superior Street, 10-220 |
| nozomu.takata@northwestern.edu | |
| Phone | 3125036066 |
| Submit Date | 2023-03-15 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-06-08 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001619 |
| Project DOI: | doi: 10.21228/M8F41P |
| Project Title: | Lactate-dependent Transcriptional Regulation Controls Mammalian Eye Morphogenesis |
| Project Summary: | Mammalian retinal metabolism favors aerobic glycolysis. However, the role of glycolytic metabolism in retinal morphogenesis remains unknown. Here we report that aerobic glycolysis is necessary for the early stages of retinal development. Taking advantage of an unbiased approach that combines the use of eye organoids and single-cell RNA sequencing we identified specific glucose transporters and glycolytic genes in retinal progenitors. Next, we determined that the optic vesicle territory of mouse embryos displays elevated levels of glycolytic activity. At the functional level, we found that removal of Glut1 (also known as Slc2a1) and Lactate dehydrogenase A (Ldha) gene activity from developing retinal progenitors arrested eye morphogenesis. Surprisingly, we uncovered that lactate-mediated upregulation of key eye-field transcription factors was controlled by the epigenetic modification of histone H3 acetylation through histone deacetylase (Hdac) activity. Our results identify a novel bioenergetic independent role of lactate as a signaling molecule necessary for mammalian eye morphogenesis. |
| Institute: | Northwestern University |
| Last Name: | TAKATA |
| First Name: | NOZOMU |
| Address: | 303 East Superior Street, 10-220, Chicago, Illinois, 60611, USA |
| Email: | nozomu.takata@northwestern.edu |
| Phone: | 13125036066 |
Subject:
| Subject ID: | SU002607 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Treatment |
|---|---|---|---|
| SA252368 | 120mins-2 | Wild-type | 120mins |
| SA252369 | 120mins-1 | Wild-type | 120mins |
| SA252370 | 15mins-1 | Wild-type | 15mins |
| SA252371 | 15mins-2 | Wild-type | 15mins |
| SA252372 | 30mins-2 | Wild-type | 30mins |
| SA252373 | 30mins-1 | Wild-type | 30mins |
| SA252374 | 60mins-1 | Wild-type | 60mins |
| SA252375 | 60mins-2 | Wild-type | 60mins |
| Showing results 1 to 8 of 8 |
Collection:
| Collection ID: | CO002600 |
| Collection Summary: | The eye organoids were collected and washed twice with PBS before pellets were flash-frozen and stored at −80°C until metabolite extraction. Three biologically independent repeated samples were collected. |
| Collection Protocol Filename: | Glucose_and_lactate_isotope_labeling.pdf |
| Sample Type: | Retina |
Treatment:
| Treatment ID: | TR002619 |
| Treatment Summary: | All the organoids (50 organoids per condition) were washed with blank SILAC before being reconstituted in 13C medium at a concentration of 10 eye organoids/ml The incubation was performed from 15 minutes to 2 hours for time-course analysis of the glucose labeling. |
Sample Preparation:
| Sampleprep ID: | SP002613 |
| Sampleprep Summary: | The eye organoids were collected and washed twice with PBS before pellets were flash-frozen and stored at −80°C until metabolite extraction. Metabolite analysis was performed as described previously (PMID: 33931446). Briefly, for metabolite extraction, 80% methanol was used followed by the rapid freeze-thaw method to break the tissues. The supernatant underwent speedvac drying. The samples were prepared in 80% acetonitrile and were analyzed by High-Resolution Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). |
Combined analysis:
| Analysis ID | AN004129 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Q-exactive |
| Column | Waters XBridge BEH Amide (100 x 3.0mm, 3.5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003059 |
| Instrument Name: | Q-exactive |
| Column Name: | Waters XBridge BEH Amide (100 x 3.0mm, 3.5um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1-18 min, 75% A; 18-25 min, 15% A with a flow rate of 150 μL/min |
| Flow Rate: | 150 μL/mi |
| Solvent A: | 95% water/5% acetonitrile; 10 mM ammonium hydroxide; 10 mM ammonium acetate, pH 9.0 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003876 |
| Analysis ID: | AN004129 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The capillary of the ESI source was set to 275 °C, with sheath gas at 35 arbitrary units, auxiliary gas at 5 arbitrary units and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, an m/z scan range from 60 to 900 was chosen and MS1 data was collected at a resolution of 70,000. The automatic gain control (AGC) target was set at 1 × 106 and the maximum injection time was 200 ms. The top 5 precursor ions were subsequently fragmented, in a data-dependent manner, using the higher energy collisional dissociation (HCD) cell set to 30% normalized collision energy in MS2 at a resolution power of 17,500. Besides matching m/z, metabolites are identified by matching either retention time with analytical standards and/or MS2 fragmentation pattern. Data acquisition and analysis were carried out by Xcalibur 4.1 software and Tracefinder 4.1 software, respectively (both from Thermo Fisher Scientific) |
| Ion Mode: | UNSPECIFIED |
