Summary of Study ST001721

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 PR001103. The data can be accessed directly via it's Project DOI: 10.21228/M84988 This work is supported by NIH grant, U2C- DK119886.

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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.

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Study IDST001721
Study TitleDetecting sex-related changes to the metabolome of a critically endangered freshwater crayfish during the mating season
Study TypeLC-MS analysis of crustacean haemolymph
Study SummaryCaptive breeding is a vital tool in the conservation of highly endangered species, as it is for the Margaret River hairy marron, Cherax tenuimanus, from the south west of Australia. A close relative, Cherax cainii, has almost completely displaced C. tenuimanus in the wild and is a successful aquaculture species, whereas C. tenuimanus has performed poorly in captivity. We used untargeted liquid chromatography-mass spectrometry to obtain metabolomic profiles of female and male C. tenuimanus held in controlled aquarium conditions during their reproductive period. Using repeated haemolymph sampling we tracked the metabolomic profiles of animals just prior to and for a period of up to 34 days after pairing with a similar sized potential mate. We identified 54 reproducible annotated metabolites including amino acids, fatty acids, biogenic amines, purine and pyrimidine metabolites and excretion metabolites. Hierarchical clustering analysis distinguished five metabolite clusters. Principal component-canonical variate analysis clearly distinguished females from males, both unpaired and paired; similar trends in profile changes in both sexes after pairing; and a striking shift in males upon pairing. We discuss three main patterns of metabolomic responses: differentiation between sexes; reactive responses to the disturbance of pairing; and convergent response to the disturbance of pairing for males. Females generally had higher concentrations of metabolites involved in metabolic rate, mobilisation of energy stores and stress. Responses to the disturbance of pairing were also related to elevated stress. Females were mobilising lipid stores to deposit yolk, whereas males had a rapid and strong response to pairing, with shifts in metabolites associated with gonad development and communication, indicating males could complete reproductive readiness only once paired with a female. The metabolomic profiles support a previously proposed potential mechanism for displacement of C. tenuimanus by C. cainii in the wild and identify several biomarkers for testing hypotheses regarding reproductive success using targeted metabolomics.
Institute
Edith Cowan University
DepartmentSchool of Science
Last NameLette
First NameEmily
Address270 Joondalup Drive, Joondalup, WA, 6027, Australia
Emaile.lette@ecu.edu.au
Phone+61 8 6304 5513
Submit Date2021-02-25
Total Subjects10
Num Males5
Num Females5
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2021-03-17
Release Version1
Emily Lette Emily Lette
https://dx.doi.org/10.21228/M84988
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN002804 AN002805
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Dionex Ultimate 3000 RS Thermo Dionex Ultimate 3000 RS
Column Thermo Hypersil GOLD C18 (100 x 2.1mm,1.9m) Thermo Hypersil GOLD C18 (100 x 2.1mm,1.9m)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units peak area values peak area values

MS:

MS ID:MS002599
Analysis ID:AN002804
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Full scans with data-dependent tandem mass spectrometry were acquired on the Orbitrap mass analyzer. Full scans were acquired at a resolution of 70,000 at mass-to-charge ratio (m/z) 200 over the m/z range 70–1000 with the ESI conditions as follows: source heater = 350°C, sheath gas = 35 (arbitrary units), auxiliary gas = 10 (arbitrary units), capillary temperature 350°C, ion spray voltage = 3.0 kV (positive ion mode) and 2.5 kV (negative ion mode), S-lens 50%, and automatic gain control = 110¬¬¬-6. Tandem mass spectrometry experiments were performed at a resolution of 17,500 at m/z 200 on each sample with the higher energy collisional dissociation energy set at 20 eV. Data acquisition was carried out using Xcalibur software (Thermo Fisher Scientific). Before analysis, the Orbitrap was externally calibrated using ready-made calibration solutions (ESI-negative ion calibration and ESI-positive ion calibration solutions) obtained from Thermo Fisher Scientific.
Ion Mode:POSITIVE
Analysis Protocol File:Methods_Lette_marron
  
MS ID:MS002600
Analysis ID:AN002805
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Full scans with data-dependent tandem mass spectrometry were acquired on the Orbitrap mass analyzer. Full scans were acquired at a resolution of 70,000 at mass-to-charge ratio (m/z) 200 over the m/z range 70–1000 with the ESI conditions as follows: source heater = 350°C, sheath gas = 35 (arbitrary units), auxiliary gas = 10 (arbitrary units), capillary temperature 350°C, ion spray voltage = 3.0 kV (positive ion mode) and 2.5 kV (negative ion mode), S-lens 50%, and automatic gain control = 110¬¬¬-6. Tandem mass spectrometry experiments were performed at a resolution of 17,500 at m/z 200 on each sample with the higher energy collisional dissociation energy set at 20 eV. Data acquisition was carried out using Xcalibur software (Thermo Fisher Scientific). Before analysis, the Orbitrap was externally calibrated using ready-made calibration solutions (ESI-negative ion calibration and ESI-positive ion calibration solutions) obtained from Thermo Fisher Scientific.
Ion Mode:NEGATIVE
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