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Absolute quantification of a steroid hormone that regulates development in Caenorhabditis elegans.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Li, Tie-Mei Chen, Jie Li, Xiangke Ding, Xiao-jun Wu, Yao Yu Zhao, Li-Feng Chen, She Lei, Xiaoguang Dong, Meng-Qiu |
| Copyright Year | 2013 |
| Abstract | Under favorable conditions, Caenorhabditis elegans larvae grow into reproductive adults after a series of molting cycles. When environmental conditions are harsh, they arrest as dauer larvae. Dafachronic acid (DA), a C. elegans steroid hormone, is required for reproductive development. Here, we report a mass spectrometry (MS) method for absolute quantitation of DA in C. elegans. The extraction of DA from C. elegans was optimized to achieve a recovery rate of greater than 83%. The MS sensitivity to DA increased 100-fold after carboxyl group derivatization with 2-picolylamine. High-resolution selected ion monitoring (HR-SIM) on a Q-Orbitrap mass spectrometer Q Exactive outperformed targeted-MS2 on the same instrument and selected reaction monitoring (SRM) on a triple-quadrupole mass spectrometer TSQ Quantum Discovery. With a limit of quantification as low as 1 pg of DA, the HR-SIM method enables absolute quantification of endogenous DA during the reproductive development of C. elegans. We found that in wild-type (WT) worms, DA increases from 0.04 ± 0.02 ng/mg protein in the L1 larval stage to 1.21 ± 0.67 ng/mg protein in the L2 larval stage and decreases again after the L3 stage. In comparison, four genetic mutants that have a constitutive dauer-formation phenotype due to disrupted insulin, TGF-β, or cGMP signaling all have a very low DA level in the L2 stage (below 15% of the WT). These mutants are able to escape the dauer fate and most of them grow into fertile adults when supplied with exogenous DA. Therefore, a DA spike in the L2 stage is critical for the reproductive development of C. elegans. |
| Starting Page | 663 |
| Ending Page | 667 |
| Page Count | 5 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://www.nibs.ac.cn/articles/20131001_02.pdf |
| PubMed reference number | 24010904v1 |
| Alternate Webpage(s) | https://doi.org/10.1021/ac402025c |
| DOI | 10.1021/ac402025c |
| Journal | Analytical chemistry |
| Volume Number | 85 |
| Issue Number | 19 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Blood Glucose Self-Monitoring Caenorhabditis elegans Carboxyl Group Cyclic GMP Fertility Ions Larva Mass Spectrometers (device) Mass Spectrometry Providing (action) Quadrupole Quantitation Reproduction Steroid hormone Steroids UNC13B gene dafachronic acid heart rate mg/kg mutant reproductive development |
| Content Type | Text |
| Resource Type | Article |
