This course provides basic introductory and comprehensive information on performing metabolic studies using tracers labeled with radioactive or
stable isotopes in humans, animals and cells. The course is designed for beginners as well as for those with experience who wish to expand their
capabilities to more sophisticated problems. The faculty has expertise in a wide variety of applications and methodologies.
Techniques are taught for the investigation of whole body metabolism, for metabolite balance across organs, intracellular flux rates and pathway
regulation in humans, animals and cells. Basic aspects of modeling will be considered, as well as specific applications to the study of carbohydrate,
fat, protein metabolism and energy balance. Theoretical and practical matters related to sample analysis by mass spectrometry and NMR are discussed,
including examples of calculations involved in determining isotopic enrichment and basic kinetic parameters. Attendees will have opportunities to
discuss their research projects in one-on-one mentoring sessions with faculty.
Course Syllabus Example:
Download 2019 Course Syllabus
Course Overview (tentative)
Monday
Basic tracer theory
Basic characteristics of radioactive isotopes
General principles of mass spectrometry
Specific activity and isotopic enrichment (GC-MS)
Methods of mass spectrometry analysis MS, LC-MS
Problem solving sessions
Tuesday
Tracer kinetics (single pool models)
Oxidation and synthesis rates
Glucose metabolism (clamp studies)
Lipid metabolism (basic kinetics)
Introduction to the NIH grant process
Practical application of the insulin clamp (breakout session)
Wednesday
Using positional isotopomer analysis to assess pathway fluxes using NMR
Protein metabolism
Thursday
Energy expenditure with doubly labeled water
Synthesis rates with deuterated water: proteins, fatty acids sterols, glucose, nucleic acids
Mass isotopomer distribution analysis: polymer synthesis, multiple flux pathways, TCA cycle, anaplerosis
Metabolic flux analysis workshop using MFA software suite
Friday
Small Group sessions
- Working with Mass Spec data
- Lipid flux
- Carbohydrate flux
- Protein flux
Pathway discovery via association of isotopomer analysis and metabolomics.
Discussion of inherently difficult problems
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