Summary of Study ST004284
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 PR002681. The data can be accessed directly via it's Project DOI: 10.21228/M83R9T 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 | ST004284 |
| Study Title | Acetyl-CoA Carboxylase depletion in germline results in increased Acetyl-CoA level |
| Study Summary | Summary of the study: Reproduction is tightly linked to nutrient availability and lipid metabolism, and disturbances in these processes often lead to reproductive failure. In our study, we investigated the metabolic mechanisms governed by Acetyl-CoA Carboxylase (ACC), a rate-limiting enzyme in fatty acid synthesis, that support oogenesis. We found that ACC modulates nutrient-responsive TOR signaling to sustain endosomal trafficking, a process essential for oocyte specification. Loss of ACC shifts cellular metabolism toward fatty acid oxidation (FAO), elevating Acetyl-CoA levels and enhancing flux through the TCA cycle and electron transport chain (ETC). This metabolic reprogramming results in hyperactivation of TOR signaling, leading to excessive protein synthesis, disrupted endosomal trafficking, and defective germ cell differentiation. |
| Institute | Institute of Cellular and Organismic Biology, Academia Sinica |
| Department | ICOB |
| Laboratory | R337, Hsu Lab |
| Last Name | Hsu |
| First Name | Hweijan |
| Address | no 128, sec 2, Academia Rd, Academia Sinica, ICOB 337, Taipei, Taipei, 11529, Taiwan |
| cohsu@gate.sinica.edu.tw | |
| Phone | 886-227871542 |
| Submit Date | 2025-09-22 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-10-16 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002681 |
| Project DOI: | doi: 10.21228/M83R9T |
| Project Title: | Acetyl-CoA Carboxylase Maintains Energetic Balance for Functional Oogenesis |
| Project Type: | MS quantitative analysis |
| Project Summary: | PROJECT SUMMARY Reproduction requires the integration of nutrient availability and lipid metabolism to sustain oogenesis and ensure fertility. Disruptions in this metabolic balance are strongly associated with reproductive failure, yet the mechanisms that coordinate these processes remain poorly defined. This project aimed to dissect how lipid metabolic pathways intersect with nutrient-sensing signals to govern germ cell differentiation. Specifically, we focused on the role of Acetyl-CoA Carboxylase (ACC), the rate-limiting enzyme for fatty acid synthesis, in maintaining oocyte development. We demonstrate that ACC functions as a key metabolic regulator by sustaining endosomal trafficking and modulating nutrient-responsive TOR signalling. Loss of ACC activity shifts metabolism toward fatty acid oxidation (FAO), which increases flux through the tricarboxylic acid (TCA) cycle and electron transport chain (ETC). This altered metabolic state hyperactivates TOR signalling, leading to excessive protein synthesis, disrupted endosomal trafficking, and impaired germ cell fate determination. Importantly, we show that these defects are reversible. Targeted interventions—including inhibition of FAO or TOR signalling, attenuation of protein synthesis, or dietary protein adjustment—restore cellular homeostasis and rescue oogenic defects. The findings from this study reveal a previously unrecognised link between lipid metabolism, nutrient-sensing pathways, and reproductive development. By defining ACC as a central coordinator of metabolic and signalling networks in oogenesis, this work advances fundamental understanding of how energy metabolism shapes reproductive outcomes. Furthermore, the identification of corrective strategies highlights potential therapeutic avenues for addressing infertility and reproductive disorders associated with metabolic imbalance. |
| Institute: | Academia Sinica |
| Department: | Institute of Cellular and Organismic Biology |
| Laboratory: | R337, Hsu Lab |
| Last Name: | Amamrtuvshin |
| First Name: | Oyundari |
| Address: | R337, ICOB, No. 128, Section 2, Academia Rd, Nangang District, Taipei City, Taiwan 115 |
| Email: | oyundariamar@gmail.com |
| Phone: | 886-227871542 |
Subject:
| Subject ID: | SU004437 |
| Subject Type: | Invertebrate |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Genotype Strain: | nos>mCherryRNAi and nos>ACCRNAi |
Factors:
Subject type: Invertebrate; Subject species: Drosophila melanogaster (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Treatment |
|---|---|---|---|
| SA498453 | AccKD1-1 | Drosophila Ovary | Acc-depleted |
| SA498454 | AccKD2-1 | Drosophila Ovary | Acc-depleted |
| SA498455 | AccKD2-2 | Drosophila Ovary | Acc-depleted |
| SA498456 | mcherryKD1-1 | Drosophila Ovary | control |
| SA498457 | mcherryKD2-1 | Drosophila Ovary | control |
| SA498458 | mcherryKD2-2 | Drosophila Ovary | control |
| Showing results 1 to 6 of 6 |
Collection:
| Collection ID: | CO004430 |
| Collection Summary: | An extraction solution (2:2:1 acetonitrile: methanol: ddH2O) was prepared and stored overnight at -20°C before use. Hundred pairs of ovaries from each genotype were dissected in cold 1xPBS. Since nos>ACC RNAi ovaries lacked vitellogenic egg chambers, the vitellogenic egg chambers from control ovaries were removed using forceps, leaving the transparent regions for metabolite extraction. Drosophila dissection was completed within 1 hour. Ovary samples for each genotype were collected in an Eppendorf tube, and metabolites from each sample (containing about 12.5 µg DNA) were extracted using 200 µL metabolite extraction solvent. Samples were homogenized by vortexing for 5 sec followed by cold bath sonication for 5 min, repeated twice. Tissue debris was pelleted by centrifugation at 15871x g (rcf) for 10 min at 4°C. A total of 200 µL metabolite-containing supernatant was transferred into a new tube. The supernatant was freeze-dried for at least 3 h using a freeze-drier (VirTis BenchTop K). Dried samples were kept at -80°C until analysed for amino acids and TCA cycle byproducts. |
| Sample Type: | Ovaries |
Treatment:
| Treatment ID: | TR004446 |
| Treatment Summary: | nosGAL4 virgins were crossed with either mCherryRNAi or AccRNAi flies. Eggs were laid and allowed to develop into adults at 18 degrees. Adult flies were then shifted to 29 degrees to increase the expression of RNAi in the germline for 7 days. The food was changed daily |
Sample Preparation:
| Sampleprep ID: | SP004443 |
| Sampleprep Summary: | Fifty pairs of ovaries from each genotype were dissected in cold 1xPBS. Since nosGAL4>ACC RNAi ovaries lacked vitellogenic egg chambers, the vitellogenic egg chambers from control ovaries were removed using forceps, leaving the transparent regions for metabolite extraction. Metabolites from each sample (containing about 12.5 µg DNA) were extracted using 200 µL metabolite extraction solvent (2:2:1 acetonitrile: methanol: ddH2O) and stored at -20°C overnight. Samples were homogenized by vortexing for 5 sec followed by cold bath sonication for 5 min, repeated twice. Tissue debris was pelleted by centrifugation at 15871x g (rcf) for 10 min at 4°C. A total of 200 µL metabolite-containing supernatant was transferred into a new tube. The supernatant was freeze-dried for at least 3 h using a freeze drier (VirTis BenchTop K). Dried samples were kept at -80°C until analyzed for amino acids and TCA cycle byproducts. |
Chromatography:
| Chromatography ID: | CH005414 |
| Chromatography Summary: | The sample was separated with ACQUITY BEH Amide column (1.7 μm particle size, 2.1 × 100 mm, Waters). The UPLC was operated at a flow rate of 0.3 mL/min and a column temperature of 40°C. The composition of mobile phase A was water containing 15 mM ammonium acetate and 0.3% ammonium hydroxide. Mobile phase B was 90% acetonitrile containing 15 mM ammonium acetate and 0.3% ammonium hydroxide. Flow gradient changes as follows: 0min-10% A (Solvent A), 8min-50% A (Solvent A), 10min-50% A (Solvent A), 11min-10% A (Solvent A), 16min-10%A (Solvent A). Characteristic MS transitions were monitored using positive multiple reaction monitoring (MRM) mode for acetyl-CoA (m/z, 810>303). Data acquisition and processing were performed using MassLynx version 4.1 and TargetLynx software (Waters Corp.). |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters ACQUITY UPLC BEH Amide (100 x 2.1 mm, 1.7 µm) |
| Column Temperature: | 40℃ |
| Flow Gradient: | 0min-10%A (Solvent A), 8min-50%A, 10min-50%A, 11min-10%A, 16min-10%A. |
| Flow Rate: | 0.3 mL/min |
| Solvent A: | 100% Water; 15 mM Ammonium acetate; 0.3% Ammonium hydroxide |
| Solvent B: | 90% Acetonitrile/10% Water; 15 mM Ammonium acetate; 0.3% Ammonium hydroxide |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN007124 |
| Analysis Type: | MS |
| Chromatography ID: | CH005414 |
| Num Factors: | 2 |
| Num Metabolites: | 1 |
| Units: | peak area |
