#METABOLOMICS WORKBENCH sbeh91Jema_20250327_162802 DATATRACK_ID:5800 STUDY_ID:ST003855 ANALYSIS_ID:AN006333 PROJECT_ID:PR002412
VERSION             	1
CREATED_ON             	April 9, 2025, 9:04 pm
#PROJECT
PR:PROJECT_TITLE                 	Mammary gland metabolism and its relevance to the fetoplacental expression of
PR:PROJECT_TITLE                 	cytokine signaling in Caveolin-1 null mice
PR:PROJECT_SUMMARY               	Mice lacking Caveolin-1 (Cav1), a major protein of the lipid raft of plasma
PR:PROJECT_SUMMARY               	membrane, show dysregulated cellular proliferation of mammary gland and an
PR:PROJECT_SUMMARY               	abnormal fetoplacental communication during pregnancy. The aim of this study is
PR:PROJECT_SUMMARY               	to better understand the functional links of mammary gland metabolism with gene
PR:PROJECT_SUMMARY               	expression of the placenta and fetus. Untargeted metabolomics analysis was
PR:PROJECT_SUMMARY               	performed to examine changes in mammary gland metabolism due to the absence of
PR:PROJECT_SUMMARY               	Cav1. The metabolomics analysis detected a total of 141 metabolites. We
PR:PROJECT_SUMMARY               	identified 81 metabolites that showed differential rank order in the level of
PR:PROJECT_SUMMARY               	expression in the mammary glands of Cav1-null compared to control mice.
PR:PROJECT_SUMMARY               	Differential metabolomics analysis identified 11 metabolites that were
PR:PROJECT_SUMMARY               	significantly changed in the mammary gland due to the absence of Cav1.
PR:PROJECT_SUMMARY               	Integrative metabolomics and transcriptomics analyses were applied to untangle
PR:PROJECT_SUMMARY               	functional links of metabolic pathways of the mammary gland with the gene
PR:PROJECT_SUMMARY               	expression changes of the placenta and fetus. The findings of this study show
PR:PROJECT_SUMMARY               	that metabolism and gene expression of the mammary gland are significantly
PR:PROJECT_SUMMARY               	impacted due to the loss of Cav1. Genes associated with specific metabolic and
PR:PROJECT_SUMMARY               	signaling pathways show coordinated expression changed in the placenta, mammary
PR:PROJECT_SUMMARY               	gland and fetal brain in Cav1-null mice. The cytokine signaling pathway emerges
PR:PROJECT_SUMMARY               	as a key player of the molecular crosstalk among the mammary gland, placenta and
PR:PROJECT_SUMMARY               	fetal brain. By interrogating the single-nuclei gene expression data of placenta
PR:PROJECT_SUMMARY               	and fetal brain previously generated from Cav1-null mice, the study further
PR:PROJECT_SUMMARY               	reveals that these metabolic and signaling genes are differentially regulated in
PR:PROJECT_SUMMARY               	specific cell types of the placenta and fetal brain. The findings of this study
PR:PROJECT_SUMMARY               	expand our understanding about the role of mammary gland metabolism in the
PR:PROJECT_SUMMARY               	regulation of fetoplacental communication in mammalian pregnancy.
PR:INSTITUTE                     	Universiy of Missouri, Columbia
PR:LAST_NAME                     	Behura
PR:FIRST_NAME                    	Susanta
PR:ADDRESS                       	920 East Campus, Columbia, Missouri, 65211, USA
PR:EMAIL                         	behuras@missouri.edu
PR:PHONE                         	573-882-1722
#STUDY
ST:STUDY_TITLE                   	Mammary gland metabolism and its relevance to the fetoplacental expression of
ST:STUDY_TITLE                   	cytokine signaling in Caveolin-1 null mice
ST:STUDY_SUMMARY                 	Mice lacking Caveolin-1 (Cav1), a major protein of the lipid raft of plasma
ST:STUDY_SUMMARY                 	membrane, show dysregulated cellular proliferation of mammary gland and an
ST:STUDY_SUMMARY                 	abnormal fetoplacental communication during pregnancy. The aim of this study is
ST:STUDY_SUMMARY                 	to better understand the functional links of mammary gland metabolism with gene
ST:STUDY_SUMMARY                 	expression of the placenta and fetus. Untargeted metabolomics analysis was
ST:STUDY_SUMMARY                 	performed to examine changes in mammary gland metabolism due to the absence of
ST:STUDY_SUMMARY                 	Cav1. The metabolomics analysis detected a total of 141 metabolites. We
ST:STUDY_SUMMARY                 	identified 81 metabolites that showed differential rank order in the level of
ST:STUDY_SUMMARY                 	expression in the mammary glands of Cav1-null compared to control mice.
ST:STUDY_SUMMARY                 	Differential metabolomics analysis identified 11 metabolites that were
ST:STUDY_SUMMARY                 	significantly changed in the mammary gland due to the absence of Cav1.
ST:STUDY_SUMMARY                 	Integrative metabolomics and transcriptomics analyses were applied to untangle
ST:STUDY_SUMMARY                 	functional links of metabolic pathways of the mammary gland with the gene
ST:STUDY_SUMMARY                 	expression changes of the placenta and fetus. The findings of this study show
ST:STUDY_SUMMARY                 	that metabolism and gene expression of the mammary gland are significantly
ST:STUDY_SUMMARY                 	impacted due to the loss of Cav1. Genes associated with specific metabolic and
ST:STUDY_SUMMARY                 	signaling pathways show coordinated expression changed in the placenta, mammary
ST:STUDY_SUMMARY                 	gland and fetal brain in Cav1-null mice. The cytokine signaling pathway emerges
ST:STUDY_SUMMARY                 	as a key player of the molecular crosstalk among the mammary gland, placenta and
ST:STUDY_SUMMARY                 	fetal brain. By interrogating the single-nuclei gene expression data of placenta
ST:STUDY_SUMMARY                 	and fetal brain previously generated from Cav1-null mice, the study further
ST:STUDY_SUMMARY                 	reveals that these metabolic and signaling genes are differentially regulated in
ST:STUDY_SUMMARY                 	specific cell types of the placenta and fetal brain. The findings of this study
ST:STUDY_SUMMARY                 	expand our understanding about the role of mammary gland metabolism in the
ST:STUDY_SUMMARY                 	regulation of fetoplacental communication in mammalian pregnancy.
ST:INSTITUTE                     	Universiy of Missouri, Columbia
ST:LAST_NAME                     	Behura
ST:FIRST_NAME                    	Susanta
ST:ADDRESS                       	920 East Campus, Columbia, Missouri, 65211, USA
ST:EMAIL                         	behuras@missouri.edu
ST:PHONE                         	573-882-1722
ST:NUM_GROUPS                    	2
ST:TOTAL_SUBJECTS                	6
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENOTYPE_STRAIN               	B6.Cg-Cav1tm1Mls/J (Cav1-null) mice; C57BL/6J (Control) mice
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	-	BLMAM-1	Sample source:Mammary gland | Genotype:C57BL6/J | Sample type:Control	RAW_FILE_NAME(Raw file name)=BLMAM-1.CDF
SUBJECT_SAMPLE_FACTORS           	-	BLMAM-2	Sample source:Mammary gland | Genotype:C57BL6/J | Sample type:Control	RAW_FILE_NAME(Raw file name)=BLMAM-2.CDF
SUBJECT_SAMPLE_FACTORS           	-	BLMAM-3	Sample source:Mammary gland | Genotype:C57BL6/J | Sample type:Control	RAW_FILE_NAME(Raw file name)=BLMAM-3.CDF
SUBJECT_SAMPLE_FACTORS           	-	CVMAM-1	Sample source:Mammary gland | Genotype:B6.Cg-Cav1tm1Mls/J | Sample type:Cav1-null	RAW_FILE_NAME(Raw file name)=CVMAMA-1.CDF
SUBJECT_SAMPLE_FACTORS           	-	CVMAM-2	Sample source:Mammary gland | Genotype:B6.Cg-Cav1tm1Mls/J | Sample type:Cav1-null	RAW_FILE_NAME(Raw file name)=CVMAMA-2.CDF
SUBJECT_SAMPLE_FACTORS           	-	CVMAM-3	Sample source:Mammary gland | Genotype:B6.Cg-Cav1tm1Mls/J | Sample type:Cav1-null	RAW_FILE_NAME(Raw file name)=CVMAMA-3.CDF
#COLLECTION
CO:COLLECTION_SUMMARY            	The B6.Cg-Cav1tm1Mls/J (Cav1-null) and control (C57BL/6J) mice were purchased
CO:COLLECTION_SUMMARY            	from the Jackson Laboratory (stock numbers: 000664 and 007083 respectively).
CO:COLLECTION_SUMMARY            	Adult male and female mice (7 weeks old) were paired in cages to induce
CO:COLLECTION_SUMMARY            	pregnancy. The females were checked for vaginal plug. The start of pregnancy
CO:COLLECTION_SUMMARY            	(day 1) was considered when a vaginal plug was observed. The control and
CO:COLLECTION_SUMMARY            	Cav1-null pregnant mice were euthanized in triplicates on gestation day 15, and
CO:COLLECTION_SUMMARY            	the mammary gland samples were dissected from mice of both the groups (n=3
CO:COLLECTION_SUMMARY            	each).
CO:SAMPLE_TYPE                   	mammary gland
#TREATMENT
TR:TREATMENT_SUMMARY             	No treatment. Genotypes B6.Cg-Cav1tm1Mls/J (Cav1-null mice) C57BL/6J (Control
TR:TREATMENT_SUMMARY             	mice) were compared.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The samples were processed as per the attached protocol of GC-MS used by the
SP:SAMPLEPREP_SUMMARY            	University of Missouri Metabolomics Center that provided the metabolomics
SP:SAMPLEPREP_SUMMARY            	service.
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Protocol_CoreLab.pdf
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Agilent 6890
CH:COLUMN_NAME                   	J&W Scientific DB-5MS (60m x 0.25mm, 0.25mm)
CH:SOLVENT_A                     	-
CH:SOLVENT_B                     	-
CH:FLOW_GRADIENT                 	-
CH:FLOW_RATE                     	1.0 mL/min
CH:COLUMN_TEMPERATURE            	Temperature program: 80C for 2 min, then ramped at 5C min-1 to 315C and held for
CH:COLUMN_TEMPERATURE            	12 min
CH:METHODS_FILENAME              	Protocol_CoreLab.pdf
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	Protocol_CoreLab.pdf
#MS
MS:INSTRUMENT_NAME               	Agilent 5973
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The spectral analysis was performed by the AMDIS (Automate Mass-spectral
MS:MS_COMMENTS                   	Deconvolution and Identification System), and metabolites were identified using
MS:MS_COMMENTS                   	a commercial NIST17 mass spectral library. The abundance of the metabolites was
MS:MS_COMMENTS                   	determined by the Metabolomics Ion-Based Data Extraction Algorithm (MET-IDEA).
MS:MS_RESULTS_FILE               	ST003855_AN006333_Results.txt	UNITS:uM	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END