#METABOLOMICS WORKBENCH efahy_20151226_121228 STUDY_ID:ST000287 ANALYSIS_ID:AN000456 PROJECT_ID:PR000229
VERSION             	1
CREATED_ON             	December 26, 2015, 12:12 pm
#PROJECT
PR:PROJECT_TITLE                 	Energetics in dystrophic muscle
PR:PROJECT_TYPE                  	Cross-sectional
PR:PROJECT_SUMMARY               	This project utilizes mouse models to establish whether lack of
PR:PROJECT_SUMMARY               	sarcolemma-localized nNOS impairs muscle metabolism and perfusion.
PR:INSTITUTE                     	University of Florida
PR:DEPARTMENT                    	Physical Therapy
PR:LABORATORY                    	Muscle Physiology Laboratory
PR:LAST_NAME                     	Forbes
PR:FIRST_NAME                    	Sean
PR:ADDRESS                       	University of Florida, College of Public Health & Health Professions, Department
PR:ADDRESS                       	of Physical Therapy, Box 100154, UFHSC
PR:EMAIL                         	scforbes@ufl.edu
PR:PHONE                         	352-273-6111
PR:FUNDING_SOURCE                	Start-up funds
#STUDY
ST:STUDY_TITLE                   	Energetics in dystrophic muscle
ST:STUDY_TYPE                    	Cross-sectional
ST:STUDY_SUMMARY                 	This project utilizes mouse models to establish whether lack of
ST:STUDY_SUMMARY                 	sarcolemma-localized nNOS impairs muscle metabolism and perfusion.
ST:INSTITUTE                     	University of Florida
ST:DEPARTMENT                    	SECIM
ST:LABORATORY                    	Muscle Physiology Laboratory
ST:LAST_NAME                     	Forbes
ST:FIRST_NAME                    	Sean
ST:ADDRESS                       	University of Florida, College of Public Health & Health Professions, Department
ST:ADDRESS                       	of Physical Therapy, Box 100154, UFHSC
ST:EMAIL                         	scforbes@ufl.edu
ST:PHONE                         	352-273-6111
ST:NUM_GROUPS                    	4
#SUBJECT
SU:SUBJECT_TYPE                  	Animal
SU:SUBJECT_SPECIES               	Mus musculus
SU:GENDER                        	Male
SU:ANIMAL_ANIMAL_SUPPLIER        	Jackson Lab
SU:ANIMAL_HOUSING                	McKnight Brain Institute, UF
SU:ANIMAL_LIGHT_CYCLE            	12 hr light/12 hr dark
SU:ANIMAL_FEED                   	ad libtum
SU:ANIMAL_WATER                  	ad libtum
SU:ANIMAL_INCLUSION_CRITERIA     	males only
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	scf_01-M1	Strain:nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_01-M2	Strain:nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_01-M3	Strain:nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_01-M4	Strain:nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_01-M5	Strain:nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_02-M1	Strain:mdx-nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_02-M2	Strain:mdx-nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_02-M3	Strain:mdx-nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_02-M4	Strain:mdx-nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_02-M5	Strain:mdx-nNOS-/-	
SUBJECT_SAMPLE_FACTORS           	-	scf_03-M1	Strain:mdx	
SUBJECT_SAMPLE_FACTORS           	-	scf_03-M2	Strain:mdx	
SUBJECT_SAMPLE_FACTORS           	-	scf_03-M3	Strain:mdx	
SUBJECT_SAMPLE_FACTORS           	-	scf_03-M4	Strain:mdx	
SUBJECT_SAMPLE_FACTORS           	-	scf_03-M5	Strain:mdx	
SUBJECT_SAMPLE_FACTORS           	-	scf_04-M1	Strain:C57Bl/10	
SUBJECT_SAMPLE_FACTORS           	-	scf_04-M2	Strain:C57Bl/10	
SUBJECT_SAMPLE_FACTORS           	-	scf_04-M3	Strain:C57Bl/10	
SUBJECT_SAMPLE_FACTORS           	-	scf_04-M4	Strain:C57Bl/10	
SUBJECT_SAMPLE_FACTORS           	-	scf_04-M5	Strain:C57Bl/10	
#COLLECTION
CO:COLLECTION_SUMMARY            	Muscles extracted under isoflurane, snap frozen with liquid nitrogen, stored at
CO:COLLECTION_SUMMARY            	-80C
CO:COLLECTION_PROTOCOL_COMMENTS  	L3-190
CO:COLLECTION_METHOD             	muscle extraction, frozen
CO:COLLECTION_LOCATION           	UF, musle physiology laboratory
CO:COLLECTION_FREQUENCY          	once/mouse
CO:COLLECTION_DURATION           	Total ~5 min
CO:COLLECTION_TIME               	no dose
CO:VOLUMEORAMOUNT_COLLECTED      	entire muscles extracted
CO:STORAGE_CONDITIONS            	-80C freezer
CO:COLLECTION_VIALS              	2 ml vials
CO:STORAGE_VIALS                 	2 ml vials
CO:COLLECTION_TUBE_TEMP          	-80C
CO:ADDITIVES                     	none
CO:BLOOD_SERUM_OR_PLASMA         	none
CO:TISSUE_CELL_IDENTIFICATION    	none
CO:TISSUE_CELL_QUANTITY_TAKEN    	none
#TREATMENT
TR:TREATMENT_SUMMARY             	No treatment
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	NAD/ATP or NADH
SP:SAMPLEPREP_PROTOCOL_FILENAME  	SB_NAD-ATP_Assay.pdf, SB_NADH_Assay_Agilent.pdf
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	NAD-ATP method
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1290 Infinity
CH:COLUMN_NAME                   	Thermo Hypercarb (50 x 2.1mm, 3um)
CH:METHODS_FILENAME              	SB_NAD-ATP_Assay_Agilent.pdf
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	NAD/ATP
MS:INSTRUMENT_NAME               	Agilent 6490 QQQ
MS:INSTRUMENT_NAME               	Agilent 6490
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	pmol/mg dry powder
MS_METABOLITE_DATA_START
Samples	scf_01-M1	scf_01-M2	scf_01-M3	scf_01-M4	scf_01-M5	scf_02-M1	scf_02-M2	scf_02-M3	scf_02-M4	scf_02-M5	scf_03-M1	scf_03-M2	scf_03-M3	scf_03-M4	scf_03-M5	scf_04-M1	scf_04-M2	scf_04-M3	scf_04-M4	scf_04-M5
Factors	Strain:nNOS-/-	Strain:nNOS-/-	Strain:nNOS-/-	Strain:nNOS-/-	Strain:nNOS-/-	Strain:mdx-nNOS-/-	Strain:mdx-nNOS-/-	Strain:mdx-nNOS-/-	Strain:mdx-nNOS-/-	Strain:mdx-nNOS-/-	Strain:mdx	Strain:mdx	Strain:mdx	Strain:mdx	Strain:mdx	Strain:C57Bl/10	Strain:C57Bl/10	Strain:C57Bl/10	Strain:C57Bl/10	Strain:C57Bl/10
NMN		2.57		3.26	3.32	3.57	2.49	3.30		5.35	3.63		3.16	3.40	5.17	2.50	2.91	2.67	2.27	2.97
NAD	1208.52	1601.82	1484.41	1837.65	1892.69	1772.16	1732.87	1157.78	1392.64	1710.87	1209.64	852.58	1478.12	1416.72	924.83	2118.29	2313.90	2318.14	2186.24	2341.62
NADP					37.08					42.88			34.52							
AMP	0.61	0.34	0.62	0.35	0.47	0.51	0.34	0.36	0.45	0.27	0.33	1.18	0.22	0.23	0.34		0.19			0.32
ADP	4.16	3.50	4.70	3.57	4.01	4.36	4.07	3.08	4.16	3.66	3.33	4.20	3.02	3.13	2.66	2.72	3.30	3.11	3.23	3.64
ATP	17.74	25.06	22.81	24.31	20.41	24.48	26.02	14.27	18.93	23.10	16.90	10.52	18.07	16.88	10.84	23.07	26.83	24.82	27.29	26.14
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Retention Index	Moverz Quant	PubChem CID
NMN	1.20	335.2, 123.1	14180
NAD	1.80	664.2, 136.1	5893
NADP	1.70	744.2, 136.1	5886
AMP	1.50	348.1, 136.1	6083
ADP	1.50	428.0, 136.1	6022
ATP	1.50	508.1, 136.1	5957
METABOLITES_END
#END