MAGE-TAB Version	1.1
Comment[MetaboBank accession]	MTBKS233
Study Title	Impact of low birthweight on intergenerational growth of the offspring in model mouse
Study Description	Low birthweight (LBW) is associated with an elevated risk of adult-onset diseases, including kidney diseases, and affects subsequent generations. However, the underlying mechanism driving these intergenerational impacts remains poorly understood. In this study, we established an animal model of LBW induced by reduced uterine perfusion pressure (RUPP) procedure that recapitulates the phenotype of LBW. The study compared the two groups of pups: those born from the sham group and the RUPP group. We analyzed metabolites in the organs (liver and placenta) of the animal model using GC-MS/MS TQ8040 (Shimadzu). The retention times indicated in the Smart Metabolites Database (Shimadzu) were used as references to create a library for data analysis. From the results, we identified relevant pathologies in adulthood and subsequent generations based on metabolic changes in the organs affected by LBW.

Experimental Design	growth condition design
Experimental Factor Name	phenotype	tissue
Experimental Factor Type	phenotype	tissue

Person Last Name	Sato	Saigusa	Sekimoto	Yamakoshi
Person First Name	Emiko	Daisuke	Akiyo	Seiko
Person Mid Initials
Person Affiliation	Tohoku University	Teikyo University	Tohoku Medical and Phermaceutical University	Tohoku University
Person Roles	submitter	submitter	submitter	submitter

PubMed ID
Publication DOI

Protocol Name	Sample collection	Extraction	Chromatography	Mass spectrometry	Data processing	Metabolite identification
Protocol Type	Sample collection	Extraction	Chromatography	Mass spectrometry	Data processing	Metabolite identification
Protocol Description	"The date of vaginal plug confirmation, an indicator of mating success, was set at 0.5 day post coitum (dpc). Female mice (ICR) were weighed daily and underwent Sham or reduced uterine perfusion pressure (RUPP) operation at 14.5 dpc with anesthetization with isoflurane. RUPP operation was performed by ligation ovarian arterioles and uterine arterioles with a 0.75 mm diameter nylon threads between them and then the nylon thread was pulled out to narrow the vessel to 0.75 mm. The laparotomy was opened for the same amount of time required for RUPP operation without stenosis, followed by suturing, which was as the Sham operation. After operation, buprenorphine hydrochloride (0.002 mg) and antibiotics were given. Immediately after operation and at 15.5 dpc, 0.5 mL of rehydration solution containing 0.3% NaCl and 4% Sucrose was administered orally. 
After operation, the pups were allowed to give birth spontaneously, and the pups of mother who had undergone Sham or RUPP operation were designated as Sham 1st generation (S1) and RUPP 1st generation (R1) groups, respectively. Spontaneously born S1 and R1 were mated with ICR male mice when they were 7~13 weeks old, and then dissected at 13.5 dpc or 18.5 dcp, placenta and liver were harvested. Samples were frozen in liquid nitrogen and stored at -80°C."	Fifty µg of placenta or liver was homogenized with 250 µL of a solution containing 55% methanol and 22% chloroform dissolved in distilled water containing 0.5 mg/mL 2-isopropylmalate (10 mL; Sigma-Aldrich). Then, dissolved in distilled water was incubated in Thermo mixier C (Eppendorf) at 37 °C with 1200 rpm shaking for 30 min. The samples were centrifuged at 4°C and 16,000 × g for 3 min. Two hundred and twenty-five µL of supernatant was collected and added to 200 µL of distilled water. The sample was dried using an evaporator under reduced pressure and lyophilized. For oximation, 20 mg/mL methoxyamine hydrochloride (80 μL; Sigma-Aldrich) dissolved in pyridine were mixed with the lyophilized sample, sonicated for 20 min, and shaken at 1,200 rpm for 90 min at 30°C. Next, 40 μL of N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA) (GL Sciences, Tokyo, Japan) were added for derivatization. The mixture was then mixed at 1,200 rpm for 30 min at 37°C, centrifuged at 16,000 × g for 5 min at 4°C, and the resulting supernatant (1 μL) was subjected to GC-MS/MS.	GC-MS/MS analysis was performed using a GC-MS/MS TQ8040 (Shimadzu) with a fused silica capillary column (BPX-5; 30 m × 0.25 mm inner diameter, film thickness: 0.25 μm; Shimadzu) and a front inlet temperature of 250°C, and a helium gas flow rate through the column of 39.0 cm/seconds. The column temperature was held at 60°C for 2 min, then raised by 15°C/min to 330°C and maintained for 3 min. The interface and ion source temperatures were 280°C and 200°C , respectively.	GC-MS/MS analysis was performed using a GC-MS/MS TQ8040 (Shimadzu). Ion source and scan polarity were electron ionization and positive, respectively. The interface and ion source temperatures were 280°C and 200°C , respectively. Data acuisition was performed by multiple reaction monitoring and data was acquired at m/z 45-600. Mass analyzer was triple quadruple.	data not submitted	data not submitted
Protocol Parameters		Post extraction;Derivatization	Chromatography instrument;Autosampler model;Column model;Column type;Guard column	Scan polarity;Scan m/z range;Instrument;Ion source;Mass analyzer
Protocol Hardware			GC-MS/MS TQ8040 (Shimadzu), BPX5 (Catalog No.: 054101, Shimadzu GLC), AOC-20i (Shimadzu)	GC-MS/MS TQ8040 (Shimadzu)
Protocol Software

Public Release Date	2023-11-24

Term Source Name
Term Source File
Term Source Version

SDRF File	MTBKS233.sdrf.txt

Comment[Study type]	targeted metabolite profiling
Comment[Experiment type]	gas chromatography-mass spectrometry
Comment[Submission type]	GC-MS
Comment[BioProject]	PRJDB17028
Comment[Related study]
Comment[Contributor]	Sato Emiko (Tohoku University), Saigusa Daisuke (Teikyo University), Akiyo Sekimoto (Tohoku Medical and Pharmaceutical University), Seiko Yamakoshi (Tohoku University)
Comment[Submission Date]	2023-11-20
Comment[Last Update Date]	2023-11-24
