#METABOLOMICS WORKBENCH hormel101_20170712_094704 DATATRACK_ID:1121 STUDY_ID:ST000792 ANALYSIS_ID:AN001263 PROJECT_ID:PR000575
VERSION             	1
CREATED_ON             	July 12, 2017, 2:45 pm
#PROJECT
PR:PROJECT_TITLE                 	Mayo Pilot and Feasibility: Targeting Myelin Metabolism to Enhance Recovery of
PR:PROJECT_TITLE                 	Function after SCI
PR:PROJECT_SUMMARY               	The loss of myelin, degeneration of the myelin producing oligodendroglia and
PR:PROJECT_SUMMARY               	impaired remyelination are essential features of traumatic spinal cord injury
PR:PROJECT_SUMMARY               	(SCI) that significantly limit patient recovery of function. The lipid rich
PR:PROJECT_SUMMARY               	composition of myelin, including exceptionally high levels of saturated fatty
PR:PROJECT_SUMMARY               	acids, underlie its essential physiological roles, including its structural and
PR:PROJECT_SUMMARY               	signaling properties and electrical insulation of axons to facilitate the
PR:PROJECT_SUMMARY               	conduction of nerve impulses. The myelin sheaths also provide metabolic support
PR:PROJECT_SUMMARY               	to the axons they wrap, and myelin health is therefore essential to the
PR:PROJECT_SUMMARY               	maintenance of axon integrity and function in the brain and spinal cord. The
PR:PROJECT_SUMMARY               	primary goal of this Pilot Proposal to the Mayo Clinic Metabolomics Core is to
PR:PROJECT_SUMMARY               	integrate highly sensitive metabolomics liquid chromatography-tandem mass
PR:PROJECT_SUMMARY               	spectrometry (LC/MS/MS) assays to quantify the lipid composition of the myelin
PR:PROJECT_SUMMARY               	membrane, with our conventional neurobehavioral approaches, enabling us to
PR:PROJECT_SUMMARY               	explore the metabolic basis of new interventions capable of promoting myelin
PR:PROJECT_SUMMARY               	regeneration and restoration of patient function. Metabolomics Core expertise in
PR:PROJECT_SUMMARY               	Magnetic Resonance Spectroscopy (NMR) based evaluation of key metabolites
PR:PROJECT_SUMMARY               	involved in CNS injury and repair (N-acetyl-L-aspartate, choline, myo-inositol,
PR:PROJECT_SUMMARY               	glucose/ glutamine and lactate) will also be applied to strengthen our
PR:PROJECT_SUMMARY               	mechanistic understanding of myelin injury and repair. Specifically, utilizing
PR:PROJECT_SUMMARY               	these innovative approaches we will test a novel hypothesis driven by new
PR:PROJECT_SUMMARY               	preliminary findings that the levels of dietary fatty acids can be optimized
PR:PROJECT_SUMMARY               	alone, or in combination with exercise training, to facilitate myelin
PR:PROJECT_SUMMARY               	regeneration and recovery of neurobehavioral function after injury to the adult
PR:PROJECT_SUMMARY               	spinal cord. In Aim 1, we will determine whether alterations in dietary fat,
PR:PROJECT_SUMMARY               	including saturated fat or omega-3 fatty acids, facilitate restoration of the
PR:PROJECT_SUMMARY               	myelin membrane and metabolite signatures of central nervous system repair after
PR:PROJECT_SUMMARY               	experimental SCI in adult mice. In Aim 2, we will determine whether exercise
PR:PROJECT_SUMMARY               	training alone or in combination with dietary fatty acid supplementation fosters
PR:PROJECT_SUMMARY               	myelin regeneration and recovery of function after experimental SCI. The
PR:PROJECT_SUMMARY               	proposed studies will leverage the expertise of the Mayo Metabolomics Core with
PR:PROJECT_SUMMARY               	that of Dr. Scarisbrick (Mayo) in myelin biology and Dr. Gomez Pinilla (UCLA) in
PR:PROJECT_SUMMARY               	central nervous system plasticity to investigate whether two highly targetable
PR:PROJECT_SUMMARY               	lifestyle variables, that is diet and exercise, can be modulated to improve
PR:PROJECT_SUMMARY               	myelin metabolism and functional recovery after SCI.
PR:INSTITUTE                     	Mayo Clinic
PR:LAST_NAME                     	Scarisbrick
PR:FIRST_NAME                    	Isobel
PR:ADDRESS                       	200 First St. SW, Rochester, Minnesota, 55905, USA
PR:EMAIL                         	scarisbrick.isobel@mayo.edu
PR:PHONE                         	507-284-0124
#STUDY
ST:STUDY_TITLE                   	Large Untargeted Profiling of Myelin to Enhance Recovery of Function after SCI
ST:STUDY_SUMMARY                 	Tissue is from adult mouse spinal cord (SC). We are submitting these samples for
ST:STUDY_SUMMARY                 	Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The
ST:STUDY_SUMMARY                 	lipid assays we request are 1) free fatty acid composition of lipids; 2) free
ST:STUDY_SUMMARY                 	fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides,
ST:STUDY_SUMMARY                 	including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is
ST:STUDY_SUMMARY                 	our top priority, followed by the lipid assays as listed. All samples were snap
ST:STUDY_SUMMARY                 	frozen at the point of harvest and approximate weights are provided. The samples
ST:STUDY_SUMMARY                 	are submitted as intact pieces of tissue. There are 20 samples total, n=5 for
ST:STUDY_SUMMARY                 	each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat
ST:STUDY_SUMMARY                 	high sucrose diet); and Keto (ketogenic diet).
ST:INSTITUTE                     	Mayo Clinic
ST:LAST_NAME                     	Scarisbrick
ST:FIRST_NAME                    	Isobel
ST:ADDRESS                       	200 First St. SW, Rochester, Minnesota, 55905, USA
ST:EMAIL                         	scarisbrick.isobel@mayo.edu
ST:PHONE                         	507-284-0124
#SUBJECT
SU:SUBJECT_TYPE                  	Mouse
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	MS6006-4	nC18-2jun16-004-r001	Group:HFHS | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-4	nC18-2jun16-004-r002	Group:HFHS | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-5	nC18-2jun16-005-r001	Group:Keto | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-5	nC18-2jun16-005-r002	Group:Keto | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-6	nC18-2jun16-006-r001	Group:LFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-6	nC18-2jun16-006-r002	Group:LFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-7	nC18-2jun16-007-r001	Group:HFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-7	nC18-2jun16-007-r002	Group:HFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-8	nC18-2jun16-008-r001	Group:HFHS | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-8	nC18-2jun16-008-r002	Group:HFHS | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-9	nC18-2jun16-009-r001	Group:Keto | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-9	nC18-2jun16-009-r002	Group:Keto | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-10	nC18-2jun16-010-r001	Group:HFHS | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-10	nC18-2jun16-010-r002	Group:HFHS | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-11	nC18-2jun16-011-r001	Group:LFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-11	nC18-2jun16-011-r002	Group:LFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-12	nC18-2jun16-012-r001	Group:HFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-12	nC18-2jun16-012-r002	Group:HFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-13	nC18-2jun16-013-r001	Group:HFHS | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-13	nC18-2jun16-013-r002	Group:HFHS | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-14	nC18-2jun16-014-r001	Group:HFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-14	nC18-2jun16-014-r002	Group:HFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-15	nC18-2jun16-015-r001	Group:Keto | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-15	nC18-2jun16-015-r002	Group:Keto | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-16	nC18-2jun16-016-r001	Group:HFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-16	nC18-2jun16-016-r002	Group:HFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-17	nC18-2jun16-017-r001	Group:LFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-17	nC18-2jun16-017-r002	Group:LFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-18	nC18-2jun16-018-r001	Group:Keto | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-18	nC18-2jun16-018-r002	Group:Keto | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-19	nC18-2jun16-019-r001	Group:HFHS | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-19	nC18-2jun16-019-r002	Group:HFHS | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-20	nC18-2jun16-020-r001	Group:LFD | rep:1	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-20	nC18-2jun16-020-r002	Group:LFD | rep:2	type=NC18
SUBJECT_SAMPLE_FACTORS           	MS6006-4	pC18-31may16-004-r001	Group:HFHS | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-4	pC18-31may16-004-r002	Group:HFHS | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-5	pC18-31may16-005-r001	Group:Keto | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-5	pC18-31may16-005-r002	Group:Keto | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-6	pC18-31may16-006-r001	Group:LFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-6	pC18-31may16-006-r002	Group:LFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-7	pC18-31may16-007-r001	Group:HFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-7	pC18-31may16-007-r002	Group:HFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-8	pC18-31may16-008-r001	Group:HFHS | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-8	pC18-31may16-008-r002	Group:HFHS | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-9	pC18-31may16-009-r001	Group:Keto | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-9	pC18-31may16-009-r002	Group:Keto | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-10	pC18-31may16-010-r001	Group:HFHS | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-10	pC18-31may16-010-r002	Group:HFHS | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-11	pC18-31may16-011-r001	Group:LFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-11	pC18-31may16-011-r002	Group:LFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-12	pC18-31may16-012-r001	Group:HFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-12	pC18-31may16-012-r002	Group:HFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-13	pC18-31may16-013-r001	Group:HFHS | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-13	pC18-31may16-013-r002	Group:HFHS | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-14	pC18-31may16-014-r001	Group:HFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-14	pC18-31may16-014-r002	Group:HFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-15	pC18-31may16-015-r001	Group:Keto | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-15	pC18-31may16-015-r002	Group:Keto | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-16	pC18-31may16-016-r001	Group:HFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-16	pC18-31may16-016-r002	Group:HFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-17	pC18-31may16-017-r001	Group:LFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-17	pC18-31may16-017-r002	Group:LFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-18	pC18-31may16-018-r001	Group:Keto | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-18	pC18-31may16-018-r002	Group:Keto | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-19	pC18-31may16-019-r001	Group:HFHS | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-19	pC18-31may16-019-r002	Group:HFHS | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-20	pC18-31may16-020-r001	Group:LFD | rep:1	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-20	pC18-31may16-020-r002	Group:LFD | rep:2	type=PC18
SUBJECT_SAMPLE_FACTORS           	MS6006-4	nhilic-1jun16-004-r001_1	Group:HFHS | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-4	nhilic-1jun16-004-r002_1	Group:HFHS | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-5	nhilic-1jun16-005-r001_1	Group:Keto | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-5	nhilic-1jun16-005-r002_1	Group:Keto | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-6	nhilic-1jun16-006-r001_1	Group:LFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-6	nhilic-1jun16-006-r002_1	Group:LFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-7	nhilic-1jun16-007-r001_1	Group:HFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-7	nhilic-1jun16-007-r002_1	Group:HFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-8	nhilic-1jun16-008-r001_1	Group:HFHS | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-8	nhilic-1jun16-008-r002_1	Group:HFHS | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-9	nhilic-1jun16-009-r001_1	Group:Keto | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-9	nhilic-1jun16-009-r002_1	Group:Keto | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-10	nhilic-1jun16-010-r001_1	Group:HFHS | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-10	nhilic-1jun16-010-r002_1	Group:HFHS | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-11	nhilic-1jun16-011-r001_1	Group:LFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-11	nhilic-1jun16-011-r002_1	Group:LFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-12	nhilic-1jun16-012-r001_1	Group:HFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-12	nhilic-1jun16-012-r002_1	Group:HFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-13	nhilic-1jun16-013-r001_1	Group:HFHS | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-13	nhilic-1jun16-013-r002_1	Group:HFHS | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-14	nhilic-1jun16-014-r001_1	Group:HFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-14	nhilic-1jun16-014-r002_1	Group:HFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-15	nhilic-1jun16-015-r001_1	Group:Keto | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-15	nhilic-1jun16-015-r002_1	Group:Keto | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-16	nhilic-1jun16-016-r001_1	Group:HFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-16	nhilic-1jun16-016-r002_1	Group:HFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-17	nhilic-1jun16-017-r001_1	Group:LFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-17	nhilic-1jun16-017-r002_1	Group:LFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-18	nhilic-1jun16-018-r001_1	Group:Keto | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-18	nhilic-1jun16-018-r002_1	Group:Keto | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-19	nhilic-1jun16-019-r001_1	Group:HFHS | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-19	nhilic-1jun16-019-r002_1	Group:HFHS | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-20	nhilic-1jun16-020-r001_1	Group:LFD | rep:1	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-20	nhilic-1jun16-020-r002_1	Group:LFD | rep:2	type=NHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-4	philic-31may16-004-r001	Group:HFHS | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-4	philic-31may16-004-r002	Group:HFHS | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-5	philic-31may16-005-r001	Group:Keto | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-5	philic-31may16-005-r002	Group:Keto | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-6	philic-31may16-006-r001	Group:LFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-6	philic-31may16-006-r002	Group:LFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-7	philic-31may16-007-r001	Group:HFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-7	philic-31may16-007-r002	Group:HFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-8	philic-31may16-008-r001	Group:HFHS | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-8	philic-31may16-008-r002	Group:HFHS | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-9	philic-31may16-009-r001	Group:Keto | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-9	philic-31may16-009-r002	Group:Keto | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-10	philic-31may16-010-r001	Group:HFHS | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-10	philic-31may16-010-r002	Group:HFHS | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-11	philic-31may16-011-r001	Group:LFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-11	philic-31may16-011-r002	Group:LFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-12	philic-31may16-012-r001	Group:HFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-12	philic-31may16-012-r002	Group:HFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-13	philic-31may16-013-r001	Group:HFHS | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-13	philic-31may16-013-r002	Group:HFHS | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-14	philic-31may16-014-r001	Group:HFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-14	philic-31may16-014-r002	Group:HFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-15	philic-31may16-015-r001	Group:Keto | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-15	philic-31may16-015-r002	Group:Keto | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-16	philic-31may16-016-r001	Group:HFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-16	philic-31may16-016-r002	Group:HFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-17	philic-31may16-017-r001	Group:LFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-17	philic-31may16-017-r002	Group:LFD | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-18	philic-31may16-018-r001	Group:Keto | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-18	philic-31may16-018-r002	Group:Keto | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-19	philic-31may16-019-r001	Group:HFHS | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-19	philic-31may16-019-r002	Group:HFHS | rep:2	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-20	philic-31may16-020-r001	Group:LFD | rep:1	type=PHILIC
SUBJECT_SAMPLE_FACTORS           	MS6006-20	philic-31may16-020-r002	Group:LFD | rep:2	type=PHILIC
#COLLECTION
CO:COLLECTION_SUMMARY            	Uninjured or SCI mice will be randomly assigned to one of four groups, LF (low
CO:COLLECTION_SUMMARY            	fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto
CO:COLLECTION_SUMMARY            	(ketogenic diet). All diets will be obtained from Research Diets, NJ USA43,45.
CO:COLLECTION_SUMMARY            	Dietary fat supplementation will be initiated at 1 week after SCI. This time
CO:COLLECTION_SUMMARY            	point for intervention was chosen to provide a meaningful timeframe for clinical
CO:COLLECTION_SUMMARY            	translation. Also, a 1 week period will allow time for mice to recover prior to
CO:COLLECTION_SUMMARY            	providing access to wheel running (Aim 2). The impact of dietary fat
CO:COLLECTION_SUMMARY            	supplementation on myelin metabolism will be examined after a period of 7 weeks,
CO:COLLECTION_SUMMARY            	including determination of (i) the lipid profile of the myelin membrane using
CO:COLLECTION_SUMMARY            	LC/MS/MS; and (ii) metabolic markers of spinal cord metabolism by NMR. Results
CO:COLLECTION_SUMMARY            	will be correlated with (iii) cellular and molecular markers of spinal cord
CO:COLLECTION_SUMMARY            	pathophysiology including the appearance of OPCs, oligodendroglia and myelin,
CO:COLLECTION_SUMMARY            	axon health, astrogliosis and inflammation; and (iv) the extent of sensorimotor
CO:COLLECTION_SUMMARY            	recovery. (v) In addition, to gauge the impact of the dietary fat on systemic
CO:COLLECTION_SUMMARY            	metabolic status, Insulin and Glucose Resistance Tests will be performed at 7
CO:COLLECTION_SUMMARY            	weeks. Food intake (g/day) and body weight gain (% initial weight) will be
CO:COLLECTION_SUMMARY            	measured daily until the endpoint of each experiment. Mice will be housed
CO:COLLECTION_SUMMARY            	individually in a temperature-controlled facility with a 12:12-h light-dark
CO:COLLECTION_SUMMARY            	cycle and ad libitum access to each diet and water. A Power Analysis was
CO:COLLECTION_SUMMARY            	performed based on histological outcomes in mice with contusion compression
CO:COLLECTION_SUMMARY            	injury. To detect a difference between groups of 20%, which would very
CO:COLLECTION_SUMMARY            	meaningful, a group size of 8 will be needed to achieve a power of 0.85. An
CO:COLLECTION_SUMMARY            	additional 2 mice per group has been added to account for mortality. The Mayo
CO:COLLECTION_SUMMARY            	Clinic Institutional Animal Care and Use Committee has approved of the proposed
CO:COLLECTION_SUMMARY            	studies.
#TREATMENT
TR:TREATMENT_SUMMARY             	To test the hypothesis that optimizing dietary fat will facilitate myelin repair
TR:TREATMENT_SUMMARY             	after SCI, the diet of uninjured adult female C57BL6/J mice (12 week, 22-25g,
TR:TREATMENT_SUMMARY             	Jackson), or those with experimental contusion-compression SCI of the
TR:TREATMENT_SUMMARY             	lumbosacral spinal cord (L2-L3) (Fejota Clip 3g Force, applied for 30s)32,47
TR:TREATMENT_SUMMARY             	will be supplemented with saturated fat. The 3g Clip produces moderate SCI
TR:TREATMENT_SUMMARY             	including demyelination and clinical impairment and we recently published a
TR:TREATMENT_SUMMARY             	detailed methodology. At 1 week after injury, the 3g injured mice are expected
TR:TREATMENT_SUMMARY             	to have an average Basso Mouse Scale score (BMS)=5 on a 9 point scale such that
TR:TREATMENT_SUMMARY             	they have frequent plantar stepping with no or some coordination. This level of
TR:TREATMENT_SUMMARY             	impairment was chosen to provide a sufficient window to observe recovery, and to
TR:TREATMENT_SUMMARY             	be at a level compatible with examination of exercise training by wheel running
TR:TREATMENT_SUMMARY             	(Aim 2).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	"large scale profiling of mouse spinal cord Lipids will be quantified in myelin
SP:SAMPLEPREP_SUMMARY            	isolated in high yield and purity by subcellular fractionation from the
SP:SAMPLEPREP_SUMMARY            	lumbosacral spinal cord. While there are no absolutely ‘myelin-specific’
SP:SAMPLEPREP_SUMMARY            	lipids, galactocerebroside is the most typical of myelin in the adult nervous
SP:SAMPLEPREP_SUMMARY            	system being directly proportional to the amount of myelin. Sulfatide is another
SP:SAMPLEPREP_SUMMARY            	galactolipid enriched in myelin. Together with cholesterol, these form 78% of
SP:SAMPLEPREP_SUMMARY            	the total amount of lipid in the myelin membrane and each will be quantified
SP:SAMPLEPREP_SUMMARY            	using LC/MS/MS. A highly sensitive assay for galactocerebroside was recently
SP:SAMPLEPREP_SUMMARY            	established by the Mayo Metabolomics Core and can be implemented immediately.
SP:SAMPLEPREP_SUMMARY            	The LC/MS/MS panel for free fatty acids, including the very long chain fatty
SP:SAMPLEPREP_SUMMARY            	acids found in myelin is also routinely performed by the Core. Cholesterol will
SP:SAMPLEPREP_SUMMARY            	be quantified using an NMR-based approach by the Mayo Dept. of Laboratory
SP:SAMPLEPREP_SUMMARY            	Medicine Clinical Core. Additionally, we have a plan in place with the
SP:SAMPLEPREP_SUMMARY            	Metabolomics Core to develop LC/MS/MS assays for sulfatide and sphingomyelin
SP:SAMPLEPREP_SUMMARY            	during the Pilot proposal. Having quantitative assays for each of these key
SP:SAMPLEPREP_SUMMARY            	myelin lipids will facilitate our goal to comprehensively profile myelin lipid
SP:SAMPLEPREP_SUMMARY            	metabolism and will form foundational assays for a future NIH grant focused on
SP:SAMPLEPREP_SUMMARY            	myelin metabolism."
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Agilent 1290 Infinity
CH:COLUMN_NAME                   	Waters Acquity HSS C18 (150 x 2.1mm, 1.8um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6550 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_RESULTS_FILE               	ST000792_AN001263_Results.txt	UNITS:intensity
#END