Summary of Study ST000965

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR000662. The data can be accessed directly via it's Project DOI: 10.21228/M83Q2T This work is supported by NIH grant, U2C- DK119886.


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Study IDST000965
Study TitleTimecourse of U13C glucose labeling of pancreatic islets in young mice
Study SummaryPancreatic islets from young mice were traced with 16.8mM U13C glucose over a 90 minute timecourse to determine the rate of metabolite labeling during glucose stimulation.
University of California, San Diego
Last NameWortham
First NameMatthew
Address2880 Torrey Pines Scenic Drive, Sanford Consortium for Regenerative Medicine, Room 3102
Submit Date2018-04-27
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailGC-MS
Release Date2019-04-27
Release Version1
Matthew Wortham Matthew Wortham application/zip

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Project ID:PR000662
Project DOI:doi: 10.21228/M83Q2T
Project Title:Integrated in vivo quantitative proteomics and nutrient tracing reveals age-related metabolic rewiring of pancreatic beta cell function
Project Summary:Aging is associated with fundamental changes in pancreatic β-cell physiology; yet, the mechanisms that drive these age-related changes are poorly understood. Here, we performed comprehensive in vivo quantitative proteomic profiling of pancreatic islets from adolescent and old mice. Nutrient tracing and targeted metabolomics demonstrated accelerated accumulation of glucose-derived metabolites and coupling factors in aged islets, indicating that age-related changes in glucose metabolism contribute to improved glucose-stimulated insulin secretion with age. Together, our study provides the first in-depth characterization of age-related changes in the islet proteome and establishes metabolic rewiring as an important mechanism for age-associated changes in β-cell function.
Institute:University of California, San Diego
Last Name:Wortham
First Name:Matthew
Address:2880 Torrey Pines Scenic Drive, Sanford Consortium for Regenerative Medicine, Room 3102, La Jolla, California, 92037, USA


Subject ID:SU001004
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL/6N
Age Or Age Range:4 weeks old (young) or 10-15 months old (old)
Gender:Male and female
Animal Animal Supplier:Charles River Labs
Animal Feed:standard chow diet (PicoLab Rodent Diet 20 pellets, #5053


Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Age Treatment
SA0577121_9_2018_islet_ugluc_timecourse_0_1Young (4 weeks) 16.8mM Glucose (0 min)
SA0577131_9_2018_islet_ugluc_timecourse_0_7Young (4 weeks) 16.8mM Glucose (0 min)
SA0577141_9_2018_islet_ugluc_timecourse_0_6Young (4 weeks) 16.8mM Glucose (0 min)
SA0577151_9_2018_islet_ugluc_timecourse_0_2Young (4 weeks) 16.8mM Glucose (0 min)
SA0577161_9_2018_islet_ugluc_timecourse_0_3Young (4 weeks) 16.8mM Glucose (0 min)
SA0577171_9_2018_islet_ugluc_timecourse_0_5Young (4 weeks) 16.8mM Glucose (0 min)
SA0577181_9_2018_islet_ugluc_timecourse_30mins_8Young (4 weeks) 16.8mM Glucose (30 min)
SA0577191_9_2018_islet_ugluc_timecourse_30mins_9Young (4 weeks) 16.8mM Glucose (30 min)
SA0577201_9_2018_islet_ugluc_timecourse_30mins_14Young (4 weeks) 16.8mM Glucose (30 min)
SA0577211_9_2018_islet_ugluc_timecourse_30mins_12Young (4 weeks) 16.8mM Glucose (30 min)
SA0577221_9_2018_islet_ugluc_timecourse_30mins_10Young (4 weeks) 16.8mM Glucose (30 min)
SA0577231_9_2018_islet_ugluc_timecourse_30mins_13Young (4 weeks) 16.8mM Glucose (30 min)
SA0577241_9_2018_islet_ugluc_timecourse_30mins_11Young (4 weeks) 16.8mM Glucose (30 min)
SA0577251_9_2018_islet_ugluc_timecourse_60mins_20Young (4 weeks) 16.8mM Glucose (60 min)
SA0577261_9_2018_islet_ugluc_timecourse_60mins_21Young (4 weeks) 16.8mM Glucose (60 min)
SA0577271_9_2018_islet_ugluc_timecourse_60mins_19Young (4 weeks) 16.8mM Glucose (60 min)
SA0577281_9_2018_islet_ugluc_timecourse_60mins_15Young (4 weeks) 16.8mM Glucose (60 min)
SA0577291_9_2018_islet_ugluc_timecourse_60mins_18Young (4 weeks) 16.8mM Glucose (60 min)
SA0577301_9_2018_islet_ugluc_timecourse_60mins_16Young (4 weeks) 16.8mM Glucose (60 min)
SA0577311_9_2018_islet_ugluc_timecourse_60mins_17Young (4 weeks) 16.8mM Glucose (60 min)
SA0577321_9_2018_islet_ugluc_timecourse_90mins_27Young (4 weeks) 16.8mM Glucose (90 min)
SA0577331_9_2018_islet_ugluc_timecourse_90mins_28Young (4 weeks) 16.8mM Glucose (90 min)
SA0577341_9_2018_islet_ugluc_timecourse_90mins_26Young (4 weeks) 16.8mM Glucose (90 min)
SA0577351_9_2018_islet_ugluc_timecourse_90mins_24Young (4 weeks) 16.8mM Glucose (90 min)
SA0577361_9_2018_islet_ugluc_timecourse_90mins_22Young (4 weeks) 16.8mM Glucose (90 min)
SA0577371_9_2018_islet_ugluc_timecourse_90mins_23Young (4 weeks) 16.8mM Glucose (90 min)
SA0577381_9_2018_islet_ugluc_timecourse_90mins_25Young (4 weeks) 16.8mM Glucose (90 min)
Showing results 1 to 27 of 27


Collection ID:CO000998
Collection Summary:Pancreata were digested using Liberase TL, and islets were purified using a Histopaque gradient followed by hand-picking under a dissecting microscope. Islets were allowed to recover overnight in RPMI 1640 media supplemented with 8 mM glucose, 10% FBS, 2 mM L-glutamine, 100 U/mL Pen/Strep, 1 mM sodium pyruvate, 10 mM HEPES, and 0.25 mg/mL amphoterecin B.
Collection Protocol Filename:Methods.pdf
Sample Type:Pancreatic Islets


Treatment ID:TR001018
Treatment Summary:Islets were starved for 1 hour in KRBH supplemented with 2.8 mM glucose as for GSIS assays. Islets were then transferred to petri dishes containing U-13C-labeled glucose as indicated and immediately seeded into wells of non-treated 12-well plates at a density of 100 size-matched islets per replicate in a final volume of 500 μl KRBH containing basal (2.8 mM) or stimulatory (16.8 mM) concentrations of labeled glucose. Tracing was performed for the indicated durations at 37oC with 5% CO2. Following the trace, islets were rapidly chilled by swirling plate to resuspend islets then transferring entire contents of each well to an Eppendorf tube on ice. Islets were then pelleted by centrifugation at 500 x g for 2 min at 4oC, washed in 1 mL of ice cold 0.9% NaCl, centrifuged as before, then pellets were frozen in a dry ice/ethanol bath and stored at -80oC.

Sample Preparation:

Sampleprep ID:SP001011
Sampleprep Summary:Metabolites were extracted from pellets using a bligh and dyer-based methanol/chloroform/water extraction with inclusion of norvaline as a polar internal standard. Briefly, 250 μl MeOH, 250 μl CHCL3, 100 μl water containing norvaline were added to pellets. This was vortexed for 10 minutes followed by centrifugation at 10,000 g for 5 minutes at 4°C. The upper phase was separated and dried under vacuum at 4°C until dry. Polar metabolites were derivatized in 2% (w/v) methoxyamine hydrochloride in pyridine and incubated at 45°C for 60 minutes. Samples were then silylated with N-tertbutyldimethylsilyl-N-methyltrifluoroacetamide (MtBSTFA) with 1% tert-butyldimethylchlorosilane (tBDMCS) at 45°C for 30 minutes. Polar derivatives were analyzed by GC-MS using a DB-35MS column (30m x 0.25 mm i.d. x 0.25 μm) installed in an Agilent 7890B gas chromatograph (GC) interfaced with an Agilent 5977A mass spectrometer (MS) with an XTR EI source using the following temperature program: 100 °C initial, increase by 3.5 °C/min to 255 °C, increase by 15 °C/min to 320 °C and hold for 3 min.

Combined analysis:

Analysis ID AN001579
Analysis type MS
Chromatography type GC
Chromatography system Agilent 7890B
Column Agilent DB5-MS (15m × 0.25mm)
MS Type EI
MS instrument type Single quadrupole
MS instrument name Agilent 5977A
Units Total ion counts


Chromatography ID:CH001108
Chromatography Summary:Polar derivatives were analyzed by GC-MS using a DB-35MS column (30m x 0.25 mm i.d. x 0.25 μm) installed in an Agilent 7890B gas chromatograph (GC) interfaced with an Agilent 5977A mass spectrometer (MS) with an XTR EI source using the following temperature program: 100 °C initial, increase by 3.5 °C/min to 255 °C, increase by 15 °C/min to 320 °C and hold for 3 min.
Instrument Name:Agilent 7890B
Column Name:Agilent DB5-MS (15m × 0.25mm)
Chromatography Type:GC


MS ID:MS001457
Analysis ID:AN001579
Instrument Name:Agilent 5977A
Instrument Type:Single quadrupole
MS Type:EI