Summary of Study ST002706

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 PR001677. The data can be accessed directly via it's Project DOI: 10.21228/M8XQ5D 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	ST002706
Study Title	Plasma metabolomics of Bmal1-KO and Bmal1-KO+AAV
Study Summary	Disruption of the circadian clock in skeletal muscle worsens local and systemic health, leading to decreased muscle strength, metabolic dysfunction, and aging-like phenotypes. Whole-body knockout mice that lack Bmal1, a key component of the molecular clock, display premature aging. Here, by using adeno-associated viruses, we rescued Bmal1 expression specifically in the skeletal muscle of Bmal1-KO mice and found that this improves their healthspan and lifespan. Plasma samples from 40-week-old KO and KO+AAV male mice were collected at ZT1 to characterize the systemic effects of muscle-specific expression of Bmal1. Overall, our findings highlight the critical role of skeletal muscle in systemic health.
Institute	University of Florida
Last Name	Esser
First Name	Karyn
Address	1345 Center Drive, M552, Gainesville, Florida, 32610, USA
Email	kaesser@ufl.edu
Phone	352-273-5728
Submit Date	2023-05-16
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2024-05-31
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP002817
Sampleprep Summary:	All provided samples were extracted following our cellular extraction procedure with pre-normalization to the sample protein content. Each extract was spiked with 1 ul of standard mixture consisting of deuterium labeled carnitine, acetylcarnitine, propionylcarnitine, butyrylcarnitine, isovalerylcarnitine, octanoylcarnitine, myristoylcarnitine and palmitoylcarnitine to targeted identification of carnitines in the sample. Metabolite extraction was done by protein precipitation using 200 µl of 8:1:1 Acetonitrile: Methanol: Acetone with 0.1% formic acid. Further protein precipitation was allowed by incubating the samples at 4°C for 20 min. Samples were placed in an ultrasonic bath for 10 min and then centrifuged at 20 000 xg for 5min at 4°C to pellet the protein. 190 µl supernatant was transferred from each sample into clean tube and dried under a gentle stream of nitrogen at 30°C. The dried extracts were re-suspended with 25 µL water with 0.1% formic acid. Resuspension was allowed at 4°C for 10 -15 min then samples were centrifuged at 20000 xg for 5 min at 4°C. Supernatants were transferred into clean LC-vials for targeted LC-MS quantitation on a Thermo Q-Exactive Oribtrap mass spectrometer with Dionex UHPLC and autosampler.

Sampleprep ID:

SP002817

Sampleprep Summary:

All provided samples were extracted following our cellular extraction procedure with pre-normalization to the sample protein content. Each extract was spiked with 1 ul of standard mixture consisting of deuterium labeled carnitine, acetylcarnitine, propionylcarnitine, butyrylcarnitine, isovalerylcarnitine, octanoylcarnitine, myristoylcarnitine and palmitoylcarnitine to targeted identification of carnitines in the sample. Metabolite extraction was done by protein precipitation using 200 µl of 8:1:1 Acetonitrile: Methanol: Acetone with 0.1% formic acid. Further protein precipitation was allowed by incubating the samples at 4°C for 20 min. Samples were placed in an ultrasonic bath for 10 min and then centrifuged at 20 000 xg for 5min at 4°C to pellet the protein. 190 µl supernatant was transferred from each sample into clean tube and dried under a gentle stream of nitrogen at 30°C. The dried extracts were re-suspended with 25 µL water with 0.1% formic acid. Resuspension was allowed at 4°C for 10 -15 min then samples were centrifuged at 20000 xg for 5 min at 4°C. Supernatants were transferred into clean LC-vials for targeted LC-MS quantitation on a Thermo Q-Exactive Oribtrap mass spectrometer with Dionex UHPLC and autosampler.