Summary of Study ST003395

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 PR002102. The data can be accessed directly via it's Project DOI: 10.21228/M8X24G 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.

Study ID	ST003395
Study Title	Short chain fatty acids analysis of Feces and Plasma in Pla2g10 knockout mice.
Study Summary	Group X Phospholipase A2 Links Colonic Lipid Homeostasis to Systemic Metabolism via Host-Microbiota Interaction. Analysis of short-chain fatty acids, including acetic acid, propionic acid, and butyric acid, in the feces and plasma of WT and Pla2g10 knockout mice that were fed either a low-fat diet (LFD), high-fat diet (HFD), lard diet, or omega-3 diet. This is a targeted analysis.
Institute	University of Tokyo
Last Name	Sato
First Name	Hiroyasu
Address	7-3-1 Hongo, Bunkyo-ku Tokyo 113-8655, Japan.
Email	sato-hr@m.u-tokyo.ac.jp
Phone	81-3-5841-1433
Submit Date	2024-08-01
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2024-08-30
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002102
Project DOI:	doi: 10.21228/M8X24G
Project Title:	Group X Phospholipase A2 Links Colonic Lipid Homeostasis to Systemic Metabolism via Host-Microbiota Interaction
Project Summary:	The gut microbiota influences physiological functions of the host, ranging from the maintenance of local gut homeostasis to systemic immunity and metabolism. Secreted phospholipase A2 group X (sPLA2-X) is abundantly expressed in colonic epithelial cells but is barely detectable in metabolic and immune tissues. Despite this distribution, sPLA2-X-deficient (Pla2g10-/-) mice displayed variable obesity-related phenotypes that were abrogated after treatment with antibiotics or cohousing with Pla2g10+/+ mice, suggesting the involvement of gut microbiota. Under housing conditions where Pla2g10-/- mice showed aggravation of diet-induced obesity and insulin resistance, they displayed increased colonic inflammation and epithelial damage, reduced production of polyunsaturated fatty acids (PUFAs) and lysophospholipids, decreased abundance of several Clostridium species, and reduced levels of short-chain fatty acids (SCFAs). These obesity-related phenotypes in Pla2g10-/- mice were reversed by dietary supplementation with omega-3 PUFAs or SCFAs. Thus, colonic sPLA2-X orchestrates omega-3 PUFA-SCFA interplay via modulation of gut microbiota, thereby secondarily affecting systemic metabolism.
Institute:	University of Tokyo
Last Name:	Sato
First Name:	Hiroyasu
Address:	Tokyo 113-8655, Japan.
Email:	sato-hr@m.u-tokyoac.jp
Phone:	81-3-5841-1433

Subject:

Subject ID:	SU003518
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Female

Factors:

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

mb_sample_id	local_sample_id	Genotype	Treatment	Sample source
SA368504	HF8	Pla2g10+/+	High fat diet	Feces
SA368505	HF22	Pla2g10+/+	High fat diet	Feces
SA368506	HF21	Pla2g10+/+	High fat diet	Feces
SA368507	HF20	Pla2g10+/+	High fat diet	Feces
SA368508	HF10	Pla2g10+/+	High fat diet	Feces
SA368509	HF9	Pla2g10+/+	High fat diet	Feces
SA368510	HF23	Pla2g10+/+	High fat diet	Feces
SA368511	HF7	Pla2g10+/+	High fat diet	Feces
SA368562	HF31	Pla2g10-/-	High fat diet	Feces
SA368563	HF32	Pla2g10-/-	High fat diet	Feces
SA368564	HF29	Pla2g10-/-	High fat diet	Feces
SA368565	HF30	Pla2g10-/-	High fat diet	Feces
SA368566	HF14	Pla2g10-/-	High fat diet	Feces
SA368567	HF13	Pla2g10-/-	High fat diet	Feces
SA368568	HF12	Pla2g10-/-	High fat diet	Feces
SA368569	HF11	Pla2g10-/-	High fat diet	Feces
SA368512	HP23	Pla2g10+/+	High fat diet	Plasma
SA368513	HP9	Pla2g10+/+	High fat diet	Plasma
SA368514	HP10	Pla2g10+/+	High fat diet	Plasma
SA368515	HP20	Pla2g10+/+	High fat diet	Plasma
SA368516	HP21	Pla2g10+/+	High fat diet	Plasma
SA368517	HP22	Pla2g10+/+	High fat diet	Plasma
SA368518	HP8	Pla2g10+/+	High fat diet	Plasma
SA368519	HP7	Pla2g10+/+	High fat diet	Plasma
SA368570	HP14	Pla2g10-/-	High fat diet	Plasma
SA368571	HP11	Pla2g10-/-	High fat diet	Plasma
SA368572	HP13	Pla2g10-/-	High fat diet	Plasma
SA368573	HP32	Pla2g10-/-	High fat diet	Plasma
SA368574	HP29	Pla2g10-/-	High fat diet	Plasma
SA368575	HP30	Pla2g10-/-	High fat diet	Plasma
SA368576	HP12	Pla2g10-/-	High fat diet	Plasma
SA368577	HP31	Pla2g10-/-	High fat diet	Plasma
SA368520	LAF21	Pla2g10+/+	Lard-rich diet	Feces
SA368521	LAF20	Pla2g10+/+	Lard-rich diet	Feces
SA368522	LAF19	Pla2g10+/+	Lard-rich diet	Feces
SA368523	LAF22	Pla2g10+/+	Lard-rich diet	Feces
SA368524	LAF3	Pla2g10+/+	Lard-rich diet	Feces
SA368525	LAF2	Pla2g10+/+	Lard-rich diet	Feces
SA368526	LAF1	Pla2g10+/+	Lard-rich diet	Feces
SA368527	LAF39	Pla2g10+/+	Lard-rich diet	Feces
SA368528	LAF38	Pla2g10+/+	Lard-rich diet	Feces
SA368529	LAF37	Pla2g10+/+	Lard-rich diet	Feces
SA368530	LAF36	Pla2g10+/+	Lard-rich diet	Feces
SA368531	LAF23	Pla2g10+/+	Lard-rich diet	Feces
SA368532	LAF4	Pla2g10+/+	Lard-rich diet	Feces
SA368578	LAF12	Pla2g10-/-	Lard-rich diet	Feces
SA368579	LAF9	Pla2g10-/-	Lard-rich diet	Feces
SA368580	LAF46	Pla2g10-/-	Lard-rich diet	Feces
SA368581	LAF45	Pla2g10-/-	Lard-rich diet	Feces
SA368582	LAF44	Pla2g10-/-	Lard-rich diet	Feces
SA368583	LAF43	Pla2g10-/-	Lard-rich diet	Feces
SA368584	LAF32	Pla2g10-/-	Lard-rich diet	Feces
SA368585	LAF31	Pla2g10-/-	Lard-rich diet	Feces
SA368586	LAF30	Pla2g10-/-	Lard-rich diet	Feces
SA368587	LAF11	Pla2g10-/-	Lard-rich diet	Feces
SA368588	LAF10	Pla2g10-/-	Lard-rich diet	Feces
SA368589	LAF29	Pla2g10-/-	Lard-rich diet	Feces
SA368533	LF2	Pla2g10+/+	Low fat diet	Feces
SA368534	LF1	Pla2g10+/+	Low fat diet	Feces
SA368535	LF17	Pla2g10+/+	Low fat diet	Feces
SA368536	LF19	Pla2g10+/+	Low fat diet	Feces
SA368537	LF3	Pla2g10+/+	Low fat diet	Feces
SA368538	LF15	Pla2g10+/+	Low fat diet	Feces
SA368539	LF16	Pla2g10+/+	Low fat diet	Feces
SA368540	LF18	Pla2g10+/+	Low fat diet	Feces
SA368590	LF27	Pla2g10-/-	Low fat diet	Feces
SA368591	LF5	Pla2g10-/-	Low fat diet	Feces
SA368592	LF28	Pla2g10-/-	Low fat diet	Feces
SA368593	LF4	Pla2g10-/-	Low fat diet	Feces
SA368594	LF6	Pla2g10-/-	Low fat diet	Feces
SA368595	LF25	Pla2g10-/-	Low fat diet	Feces
SA368596	LF26	Pla2g10-/-	Low fat diet	Feces
SA368541	LP3	Pla2g10+/+	Low fat diet	Plasma
SA368542	LP2	Pla2g10+/+	Low fat diet	Plasma
SA368543	LP19	Pla2g10+/+	Low fat diet	Plasma
SA368544	LP1	Pla2g10+/+	Low fat diet	Plasma
SA368545	LP18	Pla2g10+/+	Low fat diet	Plasma
SA368546	LP17	Pla2g10+/+	Low fat diet	Plasma
SA368547	LP16	Pla2g10+/+	Low fat diet	Plasma
SA368548	LP15	Pla2g10+/+	Low fat diet	Plasma
SA368597	LP26	Pla2g10-/-	Low fat diet	Plasma
SA368598	LP27	Pla2g10-/-	Low fat diet	Plasma
SA368599	LP28	Pla2g10-/-	Low fat diet	Plasma
SA368600	LP25	Pla2g10-/-	Low fat diet	Plasma
SA368601	LP6	Pla2g10-/-	Low fat diet	Plasma
SA368602	LP5	Pla2g10-/-	Low fat diet	Plasma
SA368603	LP4	Pla2g10-/-	Low fat diet	Plasma
SA368549	W3F6	Pla2g10+/+	w3-rich diet	Feces
SA368550	W3F7	Pla2g10+/+	w3-rich diet	Feces
SA368551	W3F8	Pla2g10+/+	w3-rich diet	Feces
SA368552	W3F5	Pla2g10+/+	w3-rich diet	Feces
SA368553	W3F25	Pla2g10+/+	w3-rich diet	Feces
SA368554	W3F26	Pla2g10+/+	w3-rich diet	Feces
SA368555	W3F27	Pla2g10+/+	w3-rich diet	Feces
SA368556	W3F28	Pla2g10+/+	w3-rich diet	Feces
SA368557	W3F29	Pla2g10+/+	w3-rich diet	Feces
SA368558	W3F40	Pla2g10+/+	w3-rich diet	Feces
SA368559	W3F41	Pla2g10+/+	w3-rich diet	Feces
SA368560	W3F42	Pla2g10+/+	w3-rich diet	Feces
SA368561	W3F24	Pla2g10+/+	w3-rich diet	Feces

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Collection:

Collection ID:	CO003511
Collection Summary:	Mice were anesthetized using isofluorane. Blood was collected by intracardiac puncture. EDTA was used as anticoagulant. Blood was centrifuged at 12,000xg for 20 mins and plasma was collected and subsequently frozen in liquid nitrogen. Samples are stored at -80°C. Allow mouse to defecate normally and collect the first two fecal pellets per animal into an tube. Samples are stored at -80°C.
Sample Type:	Feces, Plasma

Treatment:

Treatment ID:	TR003527
Treatment Summary:	Pla2g10+/+ and Pla2g10-/- mice fed a high fat diet, low fat diet, ω3 PUFA (fish oil)-rich diet or lard-rich control diet for 24 weeks.

Sample Preparation:

Sampleprep ID:	SP003525
Sampleprep Summary:	After homogenization with a bead homogenizer, the samples were centrifuged at 18,900× g for 10 min at 4 °C, and the supernatants were collected. Afterwards, the solutions were transferred to membrane filters (Nanosep 3K centrifugal device, Omega membrane; Pall Corporation) and centrifuged at 19,000× g for 20 min to remove any floating particulates in the supernatants. For analysis of mouse plasma, 380 μL of water and 20 μL of the internal standard solution were added to 200 μL of the plasma and mixed well. For derivatization of SCFAs, 20 μL of 20 mM 4-acetoamido-7-mercapto-2,1,3-benzoxadiazole (Tokyo Chemical Industry), triphenylphosphine, and 2,2’-dipyridyl disulfide (Sigma-Aldrich) in dichloromethane were added to the samples in a glass tube, and derivatization was performed at room temperature for 5 min while vortexing. The reaction solution was dried under vacuum, and then reconstituted with 200 μL of methanol.

Sampleprep ID:

SP003525

Sampleprep Summary:

After homogenization with a bead homogenizer, the samples were centrifuged at 18,900× g for 10 min at 4 °C, and the supernatants were collected. Afterwards, the solutions were transferred to membrane filters (Nanosep 3K centrifugal device, Omega membrane; Pall Corporation) and centrifuged at 19,000× g for 20 min to remove any floating particulates in the supernatants. For analysis of mouse plasma, 380 μL of water and 20 μL of the internal standard solution were added to 200 μL of the plasma and mixed well. For derivatization of SCFAs, 20 μL of 20 mM 4-acetoamido-7-mercapto-2,1,3-benzoxadiazole (Tokyo Chemical Industry), triphenylphosphine, and 2,2’-dipyridyl disulfide (Sigma-Aldrich) in dichloromethane were added to the samples in a glass tube, and derivatization was performed at room temperature for 5 min while vortexing. The reaction solution was dried under vacuum, and then reconstituted with 200 μL of methanol.

Combined analysis:

Analysis ID	AN005569
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	ExionLC
Column	Pursuit 5 C18 column (150 × 2.0mm, 5um)
MS Type	ESI
MS instrument type	Triple quadrupole
MS instrument name	ABI Sciex 6500 QTrap
Ion Mode	POSITIVE
Units	µM (for plasma) or mmol/kg (for feces)

Chromatography:

Chromatography ID:	CH004233
Instrument Name:	ExionLC
Column Name:	Pursuit 5 C18 column (150 × 2.0mm, 5um)
Column Temperature:	40°C
Flow Gradient:	30% B at 0 min, 30 to 50% B in 30 min, 50 to 30% B in 0.1 min, and 30% B in 4.9 min.
Flow Rate:	0.5 ml/min
Solvent A:	100% water; 0.1% formic acid
Solvent B:	100% acetonitrile; 0.1% formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS005294
Analysis ID:	AN005569
Instrument Name:	ABI Sciex 6500 QTrap
Instrument Type:	Triple quadrupole
MS Type:	ESI
MS Comments:	Identification was conducted using MRM transition and retention times, and quantification was performed based on the peak area of each isotope-labeled internal standard to normalize that of straight or branched SCFA having the same number of carbons.
Ion Mode:	POSITIVE