ABSTRACT
Individuals with the inborn error of metabolism, maple syrup urine disease (MSUD), are identified by newborn screening programs and treated with protein-modified diets that allow near normal growth and development. However, regardless of cause, a protein insult leads to metabolic decompensation, resulting in brain cell damage. The mechanism responsible for the damage is not well characterized due, in part, to the lack of an appropriate experimental model system with impaired branched chain alpha-ketoacid dehydrogenase (BCKD) activity. Here, we describe the construction of a rat pheochromocytoma cell (PC12) model harboring a doxycycline-controlled BCKD-kinase transgene. When BCKD-kinase is over-expressed in these cells, the endogenous BCKD activity is decreased, blocking branched chain amino acid (BCAA) catabolism. In cells over-expressing BCKD-kinase, addition of 25 mM leucine to the medium results in cell death. This experimental cell model accurately mimics the neuronal dysfunction in maple syrup urine disease and should facilitate further understanding of the pathophysiology of this disease and neuronal cell branched chain amino acid metabolism in general.
