Summary of Study ST003554

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 PR002188. The data can be accessed directly via it's Project DOI: 10.21228/M8SV5J 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 IDST003554
Study TitleInvestigation of age-dependent changes in the brain of honeybee workers using targeted metabolomics for amino acid and biogenic amine determination
Study SummaryIn the honeybee guts we measured the concentrations of 21 amino acids and 6 biogenic amines. Using a k-means clustering, the metabolites were grouped by their trends into six clusters. Tyramine and tyrosine are involved in the octopamine and dopamine biosynthesis. We found that all four metabolites were part of the same cluster, with their concentrations being significantly decreased in comparison to newly emerged workers from day 1 onwards for the entire duration of the study.
Institute
Helmholtz Centre for Environmental Research
DepartmentMolecular Systems Biology
Last NameEngelmann
First NameBeatrice
AddressPermoserstraße 15, Leipzipg, Saxony, 03418, Germany
Emailbeatrice.engelmann@ufz.de
Phone004934160251099
Submit Date2024-10-28
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2024-11-27
Release Version1
Beatrice Engelmann Beatrice Engelmann
https://dx.doi.org/10.21228/M8SV5J
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002188
Project DOI:doi: 10.21228/M8SV5J
Project Title:Comprehension of the age-dependent gut and brain interaction of honeybee workers by integration of multi omics approaches
Project Summary:In honeybees, division of labour is a key feature, with age-related behavioural transitions being closely associated with molecular changes in the brain, gut, and microbiota. In this study, to investigate these molecular changes and thus better understand their contribution to behavioural responses and modulation, we analysed the global metabolomic shifts in honeybee workers and their microbiota throughout their lives. Overall, our findings provide new insights toward developing potential biomarkers for evaluation of different functional changes related to various environmental stressors.
Institute:Helmholtz Centre for Environmental Research
Department:Molecular Systems Biology
Last Name:Engelmann
First Name:Beatrice
Address:Permoserstraße 15, Leipzipg, Saxony, 03418, Germany
Email:beatrice.engelmann@ufz.de
Phone:004934160251099

Subject:

Subject ID:SU003683
Subject Type:Insect
Subject Species:Apis mellifera
Taxonomy ID:7460
Species Group:Insects

Factors:

Subject type: Insect; Subject species: Apis mellifera (Factor headings shown in green)

mb_sample_id local_sample_id life stage age of workers in days
SA388307H2_D1_3_dilutedestablishment 1
SA388308H1_D1_3_dilutedestablishment 1
SA388309H1_D1_4_dilutedestablishment 1
SA388310H1_D1_5_dilutedestablishment 1
SA388311H2_D1_1establishment 1
SA388312H2_D1_2establishment 1
SA388313H2_D1_3establishment 1
SA388314H2_D1_4establishment 1
SA388315H2_D1_5establishment 1
SA388316H2_D1_1_dilutedestablishment 1
SA388317H2_D1_2_dilutedestablishment 1
SA388318H2_D1_5_dilutedestablishment 1
SA388319H1_D1_1_dilutedestablishment 1
SA388320H3_D1_1establishment 1
SA388321H3_D1_2establishment 1
SA388322H3_D1_3establishment 1
SA388323H3_D1_4establishment 1
SA388324H3_D1_5establishment 1
SA388325H3_D1_1_dilutedestablishment 1
SA388326H3_D1_2_dilutedestablishment 1
SA388327H3_D1_3_dilutedestablishment 1
SA388328H3_D1_4_dilutedestablishment 1
SA388329H3_D1_5_dilutedestablishment 1
SA388330H1_D1_2_dilutedestablishment 1
SA388331H2_D1_4_dilutedestablishment 1
SA388332H1_D1_5establishment 1
SA388333H1_D1_4establishment 1
SA388334H1_D1_3establishment 1
SA388335H1_D1_1establishment 1
SA388336H1_D1_2establishment 1
SA388337H2_D3_3establishment 3
SA388338H2_D3_2establishment 3
SA388339H2_D3_1establishment 3
SA388340H2_D3_3_dilutedestablishment 3
SA388341H3_D3_1_dilutedestablishment 3
SA388342H2_D3_4establishment 3
SA388343H2_D3_5establishment 3
SA388344H3_D3_2_dilutedestablishment 3
SA388345H3_D3_3_dilutedestablishment 3
SA388346H3_D3_4_dilutedestablishment 3
SA388347H3_D3_5_dilutedestablishment 3
SA388348H1_D3_2establishment 3
SA388349H1_D3_1establishment 3
SA388350H2_D3_2_dilutedestablishment 3
SA388351H2_D3_5_dilutedestablishment 3
SA388352H1_D3_2_dilutedestablishment 3
SA388353H2_D3_1_dilutedestablishment 3
SA388354H1_D3_3establishment 3
SA388355H2_D3_4_dilutedestablishment 3
SA388356H3_D3_5establishment 3
SA388357H3_D3_4establishment 3
SA388358H3_D3_3establishment 3
SA388359H3_D3_2establishment 3
SA388360H3_D3_1establishment 3
SA388361H1_D3_1_dilutedestablishment 3
SA388362H1_D3_3_dilutedestablishment 3
SA388363H1_D3_4_dilutedestablishment 3
SA388364H1_D3_5_dilutedestablishment 3
SA388365H1_D3_4establishment 3
SA388366H1_D3_5establishment 3
SA388367H2_D5_1_dilutedestablishment 5
SA388368H2_D5_5_dilutedestablishment 5
SA388369H3_D5_5establishment 5
SA388370H2_D5_4_dilutedestablishment 5
SA388371H2_D5_3_dilutedestablishment 5
SA388372H2_D5_2_dilutedestablishment 5
SA388373H2_D5_4establishment 5
SA388374H1_D5_1establishment 5
SA388375H2_D5_5establishment 5
SA388376H3_D5_4establishment 5
SA388377H2_D5_3establishment 5
SA388378H3_D5_1_dilutedestablishment 5
SA388379H3_D5_3establishment 5
SA388380H3_D5_2establishment 5
SA388381H3_D5_1establishment 5
SA388382H1_D5_1_dilutedestablishment 5
SA388383H1_D5_2_dilutedestablishment 5
SA388384H1_D5_3_dilutedestablishment 5
SA388385H1_D5_4_dilutedestablishment 5
SA388386H1_D5_5_dilutedestablishment 5
SA388387H3_D5_5_dilutedestablishment 5
SA388388H3_D5_4_dilutedestablishment 5
SA388389H3_D5_3_dilutedestablishment 5
SA388390H3_D5_2_dilutedestablishment 5
SA388391H1_D5_2establishment 5
SA388392H2_D5_2establishment 5
SA388393H1_D5_5establishment 5
SA388394H1_D5_4establishment 5
SA388395H1_D5_3establishment 5
SA388396H2_D5_1establishment 5
SA388397H3_EF_4experienced_forager >12
SA388398H3_EF_3experienced_forager >12
SA388399H3_EF_2experienced_forager >12
SA388400H3_EF_1experienced_forager >12
SA388401H2_EF_1_dilutedexperienced_forager >12
SA388402H2_EF_2_dilutedexperienced_forager >12
SA388403H2_EF_3_dilutedexperienced_forager >12
SA388404H2_EF_4_dilutedexperienced_forager >12
SA388405H2_EF_5_dilutedexperienced_forager >12
SA388406H3_EF_5experienced_forager >12
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Collection:

Collection ID:CO003676
Collection Summary:Workers were pooled to obtain a final sample number of n = 5. The brain(s) and gut(s) from the same workers were assigned the same sample names. In total, we had 15 samples from each age group, with the exception of the foraging stages, where owing to the lack of foragers from Hive 1, we only had 11. All sampled bees were placed directly in liquid nitrogen to prevent changes in the metabolic profile. They were then decapitated in the laboratory so that the head and body of each bee were kept in a separate Eppendorf tube (Eppendorf, Germany) for later dissection. Honeybee brains were dissected using a dissecting microscope (SZX7, Olympus). The head was placed on a petri dish lined with SYLGARD® 184 silicone. Before pinning the head for dissection, antennae and mouthpieces were removed. Subsequently, double-distilled water (ddH2O) was added until the head was fully submerged to facilitate dissection. The head cuticle was removed, and the hypopharyngeal glands and trachea were extracted from the inside. The brain was then separated from the cuticle and remaining tracheal remnants were removed before transferring the clean brain to a new Eppendorf tube.
Sample Type:Bee brain

Treatment:

Treatment ID:TR003692
Treatment Summary:On the day of newly emerged workers (day 0), bees were samples as a baseline for no or low bacterial abundance in the gut. Moreover, bees from each hive were collected on Day 1, 3 and 5 to cover the establishment phase of the gut. Samples for the in-hive worker phase, defined as workers inside the hives that have a fully developed microbiome, were taken every day from Day 7 to Day 10. For the foraging phase, we differentiated between new and experienced foragers, as these show different behaviour due to different levels of experience.Based on experience, they were either collected one to two days after onset (new foragers) or at least four days after onset (experienced foragers).

Sample Preparation:

Sampleprep ID:SP003690
Sampleprep Summary:In brief, x mg pool brain was mixed with five times the volume (in µL) of acetonitrile (ACN):Water (1:1, v/v) and homogenized using a TissueLyser II (30 Hz, 10 min; Retsch Qiagen). After Centrifugation (2 min, 14000 rpm), 10 µL were used for amino acid derivatization. First, the supernatant was evaporated to dryness (SpeedVac, Eppendorf), resuspended in 50 µL of 5% phenyl isothiocyanate (PITC) in ethanol: H2O: pyridine (1:1:1, v/v/v), and incubated for 25 min at RT. Subsequently, the samples were dried to remove excess PITC and resuspended in 10 µL 5 mM ammonium acetate in methanol. After incubation (10 min at 14,000 rpm) 90 µL of H2O: ACN + 0.2% formic acid were added. Each derivative was measured in an undiluted and diluted (1:25) manner.

Chromatography:

Chromatography ID:CH004437
Instrument Name:Waters Acquity
Column Name:Zorbax Eclipse XDB-C18 (100 x 3.0mm, 3.5um)
Column Temperature:50
Flow Gradient:0-0.5 min at 0% B, 0.5-4 min 0-70% B, 4-5.3 min 70% B, 5.3-5.4 min 70-0% B, 5.4-7.3 min 0% B
Flow Rate:0.5 mL/min
Solvent A:100% water; 0.2% formic acid
Solvent B:100% acetonitrile; 0.2% formic acid
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN005842
Analysis Type:MS
Chromatography ID:CH004437
Num Factors:7
Num Metabolites:21
Units:µMol
  
Analysis ID:AN005843
Analysis Type:MS
Chromatography ID:CH004437
Num Factors:7
Num Metabolites:6
Units:µMol
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