Summary of Study ST003473
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 PR002132. The data can be accessed directly via it's Project DOI: 10.21228/M81N7D 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.
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| Study ID | ST003473 |
| Study Title | 1H NMR metabolomics applied to assess the direct and transgenerational effects of simvastatin on the metabolism of the amphipod Gammarus locusta |
| Study Type | 1H NMR metabolomics to study the metabolic effects of direct exposure to simvastatin and transgenerational effects on the polar metabolome of amphipod Gammarus locusta |
| Study Summary | In this study, a comprehensive untargeted 1H NMR metabolomics strategy was applied to measure the metabolic impact of direct and transgenerational exposure (F0 and F3 generations, respectively) of amphipods G. locusta to simvastatin (SIM), a pharmaceutical widely prescribed for the treatment of hypercholesterolemia. To assess the direct and transgenerational effects of exposure to SIM on the metabolism of G. locusta, the following conditions were studied: i) direct exposure of parental generation (F0) of males and females, GM/GF-F0 (Exp); and ii) transgenerational effects, considering the first unexposed generation (F3) of males and females, to assess transgenerational effects, GM/GF-F3 (Tr). The obtained data added important knowledge, unveiling individual metabolic effects of direct exposure to simvastatin and its transgenerational effects, potentially contributing towards improving hazard and risk assessment of biologically active compounds. |
| Institute | University of Aveiro |
| Department | CICECO – Aveiro Institute of Materials, Department of Chemistry |
| Laboratory | Metabolomics Group |
| Last Name | Rodrigues |
| First Name | João A. |
| Address | University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal |
| joao.rodrigues@ua.pt | |
| Phone | 00351234370707 |
| Submit Date | 2024-08-19 |
| Num Groups | 8 |
| Total Subjects | 39 |
| Num Males | 20 |
| Num Females | 19 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | fid |
| Analysis Type Detail | NMR |
| Release Date | 2025-01-12 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002132 |
| Project DOI: | doi: 10.21228/M81N7D |
| Project Title: | 1H NMR metabolomics applied to assess the direct and transgenerational effects of simvastatin on the metabolism of the amphipod Gammarus locusta |
| Project Type: | 1H NMR metabolomics to study the metabolic effects of direct exposure to simvastatin and transgenerational effects on the polar metabolome of amphipod Gammarus locusta |
| Project Summary: | Pharmaceutical compounds (PhACs) and their metabolites are considered contaminants of emerging concern (CECs) to public and environmental health, due to their high stability, bioactivity, and persistence in conventional wastewater treatment plants. Aquatic organisms can be chronically exposed to PhACs during critical periods of their life or even through multi-generations. This may result in severe physiological/metabolic/endocrine disturbances, not only through direct exposure but also through inter- and transgenerational inheritance (in the absence of the insult). Special attention is given to simvastatin, one of the PhACs most prescribed to humans for the primary treatment of hypercholesterolemia, known to affect endocrine functions and disrupt reproduction, development, neuronal processes and/or other important physiological responses; not only in exposed individuals, but also in subsequent non-exposed generations. Previous studies observed that direct exposure and transgenerational exposure to simvastatin has been found to impact severely on crustaceans’ reproduction, growth and development, although the underlying mechanism remains partially unclear. NMR metabolomic approaches may provide crucial complementary mechanistic information about the cascade of metabolic events occurring with exposure. In this study a comprehensive untargetd 1H NMR metabolomics study was applied to measure the metabolic effects of direct exposure to environmentally relevant concentrations of simvastatin (F0) and transgenerational exposure (F3, where only the F0 generation was exposed) on the keystone marine amphipod species Gammarus locusta. Furthermore, NMR is here employed for the first time to address G. locusta metabolic behavior. The obtained data added important knowledge, paving the way to an improved understanding of the metabolic events cascade associated with simvastatin exposure. |
| Institute: | University of Aveiro |
| Department: | CICECO – Aveiro Institute of Materials, Department of Chemistry |
| Laboratory: | Metabolomics Group |
| Last Name: | Rodrigues |
| First Name: | Joao A. |
| Address: | University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal |
| Email: | joao.rodrigues@ua.pt |
| Phone: | 00351234370707 |
| Funding Source: | This work was developed within TRANSEPIC − Exploring Transgenerational Epigenetic Inheritance: New Methods and Strategies to Improve Environmental Hazard and Risk Assessment of Key Contaminants of Emerging Concern (CECs) [Reference: 2022.02922.PTDC, doi: 10.54499/2022.02922.PTDC], financed by the Portuguese Foundation for Science and Technology (FCT). This work was also developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 (doi: 10.54499/UIDB/50011/2020), UIDB/50011/2020, UIDP/50011/2020 (doi: 10.54499/UIDP/50011/2020) & LA/P/0006/2020, financed by national funds through the FCT/MCTES (PIDDAC). The NMR spectrometer is part of the National NMR Network (PTNMR) and are partially supported by Infrastructure Project Nº 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). T.N. acknowledges FCT Individual Call to Scientific Employment Stimulus 2022 (2022.02925.CEECIND/CP1728/CT0004, doi: 10.54499/ 2022.02925.CEECIND/CP1728/CT0004). |
Subject:
| Subject ID: | SU003601 |
| Subject Type: | Other organism |
| Subject Species: | Gammarus locusta |
| Taxonomy ID: | 65426 |
| Weight Or Weight Range: | length: 14-16 and 10-12.5 mm for males and females, respectively (whole body) |
| Gender: | Male and female |
| Species Group: | Other |
Factors:
Subject type: Other organism; Subject species: Gammarus locusta (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Factor |
|---|---|---|---|
| SA383414 | GF-F0_Ctr_05 | whole_body | GF-F0_Ctr |
| SA383415 | GF-F0_Ctr_02 | whole_body | GF-F0_Ctr |
| SA383416 | GF-F0_Ctr_01 | whole_body | GF-F0_Ctr |
| SA383417 | GF-F0_Ctr_04 | whole_body | GF-F0_Ctr |
| SA383418 | GF-F0_Ctr_03 | whole_body | GF-F0_Ctr |
| SA383419 | GF-F0_Exp_07 | whole_body | GF-F0_Exp |
| SA383420 | GF-F0_Exp_08 | whole_body | GF-F0_Exp |
| SA383421 | GF-F0_Exp_09 | whole_body | GF-F0_Exp |
| SA383422 | GF-F0_Exp_10 | whole_body | GF-F0_Exp |
| SA383423 | GF-F0_Exp_06 | whole_body | GF-F0_Exp |
| SA383424 | GF-F3_Ctr_01 | whole_body | GF-F3_Ctr |
| SA383425 | GF-F3_Ctr_02 | whole_body | GF-F3_Ctr |
| SA383426 | GF-F3_Ctr_04 | whole_body | GF-F3_Ctr |
| SA383427 | GF-F3_Ctr_03 | whole_body | GF-F3_Ctr |
| SA383428 | GF-F3_Tr_06 | whole_body | GF-F3_Tr |
| SA383429 | GF-F3_Tr_07 | whole_body | GF-F3_Tr |
| SA383430 | GF-F3_Tr_08 | whole_body | GF-F3_Tr |
| SA383431 | GF-F3_Tr_09 | whole_body | GF-F3_Tr |
| SA383432 | GF-F3_Tr_10 | whole_body | GF-F3_Tr |
| SA383433 | GM-F0_Ctr_05 | whole_body | GM-F0_Ctr |
| SA383434 | GM-F0_Ctr_04 | whole_body | GM-F0_Ctr |
| SA383435 | GM-F0_Ctr_03 | whole_body | GM-F0_Ctr |
| SA383436 | GM-F0_Ctr_01 | whole_body | GM-F0_Ctr |
| SA383437 | GM-F0_Ctr_02 | whole_body | GM-F0_Ctr |
| SA383438 | GM-F0_Exp_07 | whole_body | GM-F0_Exp |
| SA383439 | GM-F0_Exp_08 | whole_body | GM-F0_Exp |
| SA383440 | GM-F0_Exp_09 | whole_body | GM-F0_Exp |
| SA383441 | GM-F0_Exp_10 | whole_body | GM-F0_Exp |
| SA383442 | GM-F0_Exp_06 | whole_body | GM-F0_Exp |
| SA383443 | GM-F3_Ctr_01 | whole_body | GM-F3_Ctr |
| SA383444 | GM-F3_Ctr_02 | whole_body | GM-F3_Ctr |
| SA383445 | GM-F3_Ctr_04 | whole_body | GM-F3_Ctr |
| SA383446 | GM-F3_Ctr_05 | whole_body | GM-F3_Ctr |
| SA383447 | GM-F3_Ctr_03 | whole_body | GM-F3_Ctr |
| SA383448 | GM-F3_Tr_06 | whole_body | GM-F3_Tr |
| SA383449 | GM-F3_Tr_07 | whole_body | GM-F3_Tr |
| SA383450 | GM-F3_Tr_08 | whole_body | GM-F3_Tr |
| SA383451 | GM-F3_Tr_09 | whole_body | GM-F3_Tr |
| SA383452 | GM-F3_Tr_10 | whole_body | GM-F3_Tr |
| Showing results 1 to 39 of 39 |
Collection:
| Collection ID: | CO003594 |
| Collection Summary: | To assess the direct and transgenerational effects of exposure to SIM on the metabolism of G. locusta, the following conditions were studied: i) direct exposure of parental generation (F0) of males and females, GM/GF-F0 (Exp); and ii) transgenerational effects, considering the first unexposed generation (F3) of males and females, to assess transgenerational effects, GM/GF-F3 (Tr). The experimental protocol followed an earlier report (Neuparth et al., 2020). Direct exposure (F0 generation) experiments involved the selection of 50 one-week-old G. locusta offspring randomly placed in 7 L aquaria. Each aquarium was filled with natural filtered seawater (5 L) defining the following two groups: i) exposed to a SIM (320 ng/L) solution in 0.0005% acetone, GM/GF-F0 (Exp) group; and ii) controls, in filtered natural seawater with 0.0005% acetone, GM/GF-F0 (Ct) group. The chosen SIM concentration (320 ng/L) was based on concentrations found/estimated in Portuguese wastewaters. In each aquarium, a sediment layer of ca. 1 cm and several small stones were placed to simulate the species’ natural habitat. G. locusta individuals were kept in filtered natural saltwater (33–35‰ salinity), at 20 °C, under a 16:8h (light: dark) photoperiod, and were fed ad libitum with the macroalgae Ulva sp.. The water was renewed every 3 days, with the SIM concentration being re-established by spiking each aquarium with a standard solution. The actual SIM concentration was monitored during the assay. G. locusta individuals were kept under the conditions above for 55 days (until adulthood), after which 5 males (GM) and 5 females (GF) were randomly collected (whole body, 14-16 and 10-12.5 mm in length, for males and females respectively) from each condition and stored at - 80°C until tissue extraction. To evaluate the transgenerational effects of SIM exposure, the offspring of the F0 generation organisms (exposed to SIM) were placed in SIM-free water (natural filtered seawater) during three consecutive generations (F1, F2 and F3), giving rise to the group designated as GM/GF-F3 (Tr) (Figure 1). The corresponding control group comprised the F3 generation of the individuals previously unexposed to SIM, designated as GM/GF-F3 (Ct). For each condition, 4 replicate aquaria were used. Each generation was started with 50 offspring of the previous one, and kept for 55-65 days, to achieve adulthood. At the F3 generation, 5 males (GM) and 5 females (GF) (measuring 14-16 and 10-12.5 mm in length, respectively) were randomly collected for transgenerational, F3 (Tr), and control groups, F3 (Ct), and stored at -80°C until tissue extraction. |
| Collection Protocol Filename: | 1. Gammarus experimental procedure |
| Sample Type: | Whole animals |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003610 |
| Treatment Summary: | To assess the direct and transgenerational effects of exposure to SIM on the metabolism of G. locusta, the following conditions were studied: i) direct exposure of parental generation (F0) of males and females, GM/GF-F0 (Exp); and ii) transgenerational effects, considering the first unexposed generation (F3) of males and females, to assess transgenerational effects, GM/GF-F3 (Tr). Direct exposure (F0 generation) experiments involved the selection of 50 one-week-old G. locusta offspring randomly placed in 7 L aquaria. Each aquarium was filled with natural filtered seawater (5 L) defining the following two groups: i) exposed to a SIM (320 ng/L) solution in 0.0005% acetone, GM/GF-F0 (Exp) group; and ii) controls, in filtered natural seawater with 0.0005% acetone, GM/GF-F0 (Ct) group. The chosen SIM concentration (320 ng/L) was based on concentrations found/estimated in Portuguese wastewaters. In each aquarium, a sediment layer of ca. 1 cm and several small stones were placed to simulate the species’ natural habitat. G. locusta individuals were kept in filtered natural saltwater (33–35‰ salinity), at 20 °C, under a 16:8h (light: dark) photoperiod, and were fed ad libitum with the macroalgae Ulva sp.. The water was renewed every 3 days, with the SIM concentration being re-established by spiking each aquarium with a standard solution. The actual SIM concentration was monitored during the assay. G. locusta individuals were kept under the conditions above for 55 days (until adulthood), after which 5 males (GM) and 5 females (GF) were randomly collected (whole body, 14-16 and 10-12.5 mm in length, for males and females respectively) from each condition and stored at - 80°C until tissue extraction. To evaluate the transgenerational effects of SIM exposure, the offspring of the F0 generation organisms (exposed to SIM) were placed in SIM-free water (natural filtered seawater) during three consecutive generations (F1, F2 and F3), giving rise to the group designated as GM/GF-F3 (Tr) (Figure 1). The corresponding control group comprised the F3 generation of the individuals previously unexposed to SIM, designated as GM/GF-F3 (Ct). For each condition, 4 replicate aquaria were used. Each generation was started with 50 offspring of the previous one, and kept for 55-65 days, to achieve adulthood. At the F3 generation, 5 males (GM) and 5 females (GF) (measuring 14-16 and 10-12.5 mm in length, respectively) were randomly collected for transgenerational, F3 (Tr), and control groups, F3 (Ct), and stored at -80°C until tissue extraction. |
| Treatment Protocol Filename: | 1. Gammarus experimental procedure |
| Treatment Compound: | Direct (F0 generation) and transgenerational (F3 generation) exposure to simvastatin |
| Treatment Dose: | G. locusta animals were: i) directly exposed to 320 ng/L of simvastatin for 55 days (until adulthood); and ii) transgenerational exposed to simvastatin by the offspring of the F0 generation organisms being exposed to 320 ng/L of simvastatin for 55 days (F0 generations) and the following generations were placed in a simvastatin-free water (natural filtered seawater) during three consecutive generations (F1, F2 and F3). |
| Treatment Doseduration: | 55 days for direct exposure; 220 days for transgenerational exposure |
Sample Preparation:
| Sampleprep ID: | SP003608 |
| Sampleprep Summary: | Metabolite extraction was carried out using a water/methanol/chloroform method, 0.75:1:1 (v/v/v), as described elsewhere (Hines, Oladiran, Bignell et al., 2007). Briefly, each frozen G. locusta individual was placed in 2 mL Precellys tubes with 150 mg glass beads (ø = 0.5 mm). A volume of 750 μL of cold methanol: ultrapure water (4:1) solution was added before homogenization with a Precellys Evolution Touch homogenizer (Bertin Technologies, Montigny-le-Bretonneux, France) (two cycles of 10 s homogenization at 6500 rpm, with 15 s of rest between cycles). Subsequently, 300 µL cold chloroform was added and samples were vortexed, followed by addition of 300 µL cold chloroform and 300 µL cold ultrapure water. After 10 min at -20 °C, samples were centrifuged (2,500 g, 4 °C, 10 min) and the upper (polar) layer was transferred into vials, dried in a centrifugal vacuum concentrator (UNIVAP 100H) and stored at -80 °C until NMR analysis. |
| Sampleprep Protocol Filename: | 1. Gammarus experimental procedure |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | Water/methanol/chloroform method, as described in (Hines, Oladiran, Bignell et al., 2007) |
| Extract Storage: | -80℃ |
| Sample Resuspension: | The dried polar extracts of G. locusta samples were re-suspended in 600 μL of 100 mM sodium phosphate buffer (pH 7.4), in D2O (99.9% deuterium) containing 0.5 mM sodium salt of 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP-d4, in D2O, for chemical shift referencing). After vortex homogenization and centrifugation (16,000 g, room temperature, 10 min), 550 μL of the supernatant were transferred to 5 mm NMR tubes. |
| Sample Spiking: | 0.5 mM sodium salt of 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP-d4), as a chemical shift reference. |
Analysis:
| Analysis ID: | AN005708 |
| Laboratory Name: | Metabolomics Group |
| Analysis Type: | NMR |
| Software Version: | TopSpin3.2 and Amix3.9.14 |
| Operator Name: | Joao A. Rodrigues |
| Detector Type: | NMR Bruker Avance III 500 MHz |
| Results File: | ST003473_AN005708_Results.txt |
| Units: | Intensity |
NMR:
| NMR ID: | NM000290 |
| Analysis ID: | AN005708 |
| Instrument Name: | Bruker Avance III |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D-1H |
| Spectrometer Frequency: | 500 MHz |
| NMR Probe: | TXI |
| NMR Solvent: | 100 mM sodium phosphate buffer (pH 7.4), in D2O (99.9% deuterium) containing 0.5 mM sodium salt of 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP-d4, in D2O, for chemical shift referencing). |
| NMR Tube Size: | 5 mm NMR tubes |
| Shimming Method: | Topshim |
| Pulse Sequence: | noesypr1d |
| Water Suppression: | presat |
| Pulse Width: | 90-degree |
| Receiver Gain: | 203 |
| Offset Frequency: | 2352 Hz |
| Chemical Shift Ref Cpd: | 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP-d4) |
| Temperature: | 298 K |
| Number Of Scans: | 256 scans |
| Dummy Scans: | 8 |
| Acquisition Time: | 2.33 s |
| Relaxation Delay: | 5 s |
| Spectral Width: | 7,002.8 Hz |
| Num Data Points Acquired: | 32 k points |
| Line Broadening: | 0.3 Hz |
| Zero Filling: | 64 k points |
| Apodization: | Exponential |
| Baseline Correction Method: | Manual |
| Chemical Shift Ref Std: | 0 ppm for TSP-d4 |
