7th Annual MMPC Course: Isotope Tracers in Metabolic Research

Event Details

Sponsored by NIDDK and organized on behalf of the NIH-sponsored Mouse Metabolic Phenotyping Centers (MMPC)

This is the seventh annual offering of a course that provides basic introductory and comprehensive information on performing metabolic studies using tracers labeled with radioactive or stable isotopes, in man and in animals. The course is designed for beginners as well as those with experience who wish to expand their capabilities to more sophisticated problems. The faculty is well-versed in a variety of applications and methodologies.

Techniques will be presented for investigating whole body metabolism, for metabolite balance across organs, intracellular flux rates and pathway regulation. The basic aspects of modeling will be considered, as well as specific applications in the study of carbohydrate, fat, protein metabolism and energy balance. Theoretical and practical matters related to sample analysis by mass spectrometry and NMR will be discussed, including detailed numerical examples of calculations involved in determining isotopic enrichment and basic kinetic parameters. Advanced lectures will discuss in more detail the use of positional and mass isotopomer analysis for intracellular flux rates and various aspects of protein and amino acid metabolism. Applications in humans and animal models (particularly mouse) will be considered.

Course material will be available for download from http://www.mmpc.org/shared/tracers.aspx

Problems and discussion questions will highlight key concepts. In addition to organized sessions, individual attendees will have ample opportunities for personal interaction with faculty members in the form of one-on-one mentoring sessions to discuss their research projects in more depth.

Agenda

November 10, 2014

Basic characteristics of radioactive, stable isotope tracers.

General principles of mass spectrometry.
Isotopic enrichment using GC-MS.

Methods of mass spectrometry analysis.
Measurement of specific activity.

November 11, 2014

Tracer kinetics (single pool models).
Oxidation and synthesis rates.

Glucose metabolism (clamp studies).
Lipid metabolism (basic kinetics).

November 12, 2014

Pathway fluxes using NMR isotopomer analysis.
Methods in protein metabolism.

November 13, 2014

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.

November 14, 2014

Pathway discovery via association of isotopomer analysis and metabolomics.
Inherently difficult problems.