Cataplerotic TCA cycle flux determined as glutamate-sustained oxygen consumption in primary cultures of astrocytes

Utilization of glucose by adult brain as its metabolic substrate does not mean that glutamate cannot be synthesized from glucose and subsequently oxidatively degraded. Between 10 and 20% of total pyruvate metabolism in brain occurs as formation of oxaloacetate (OAA), a tricarboxylic acid (TCA) cycle intermediate, from pyruvate plus CO(2). This anaplerotic ('pool-filling') process occurs in astrocytes, which in contrast to neurons express pyruvate carboxylase (PC) activity. Equivalent amounts of pyruvate are converted to acetylcoenzyme A and condensed with oxaloacetate to form citrate (Cit), which is metabolized to alpha-ketoglutarate (generating oxidatively-derived energy), glutamate and glutamine and transferred to neurons in the glutamate-glutamine cycle and used as precursor for transmitter glutamate. Since the blood-brain barrier is poorly permeable to glutamate and its metabolites, net synthesis of glutamate must be followed by degradation of equivalent amounts of glutamate, a cataplerotic ('pool-emptying') process, in which glutamate is converted in the TCA cycle to malate or oxaloacetate (generating additional energy), which exit the cycle to form one molecule pyruvate. To obtain an estimate of the rate of astrocytic oxidation of glutamate the rate of oxygen consumption was measured in primary cultures of mouse astrocytes metabolizing glutamate in the absence of other metabolic substrates. The observed rate is compatible with complete oxidative degradation of glutamate.