ABSTRACT
Piperidine is actively transported into the synaptosomal fraction of adult mouse brain. The transport mechanism appears to be Na+ independent but is temperature dependent and sensitive to ouabain. Analysis of kinetic experiments indicates only a "low-affinity" transport system to be present. By contrast the uptake of D,L-[(3)H]pipecolic acid at a concentration of 4 X 10(-7)M was temperature and Na+ dependent, ouabain sensitive, and revealed a two-component system with a Km = 3.9 plus or minus 0.17 X 10(-6)M, Vmax = 129 plus or minus 6 pmol/mg protein/3 min for the "high-affinity" system and a Km = 90.2 plus or minus 4.3 X 10 (-6)M, Vmax = 2.45 plus or minus 0.19 nmol/mg protein/3 min for the "low-affinity" system. Compounds structurally related to pipecolic acid such as glycine, L-proline, 4-amino-n-butyric acid, and 5-amino-n-valeric acid showed an inhibitory effect on uptake at a concentration of 10(-4)M. The demonstration of biosynthesis of pipecolic acid in mouse brain and the presence of a "high-affinity" sodium-dependent uptake system suggest a physiological role of this substance in the central nervous system.
Subject(s)
Brain/metabolism , Pipecolic Acids/metabolism , Piperidines/metabolism , Animals , Biological Transport/drug effects , Female , Male , Mice , Mice, Inbred C57BL , Ouabain/pharmacology , Pipecolic Acids/biosynthesis , Sodium/pharmacology , Structure-Activity Relationship , Synaptosomes/metabolismABSTRACT
L-Pipecolate formation exhibits considerable regional differences in the central nervous system of the mouse, dog, and monkey, as reflected in measurements of the activity of delta1-pyrroline-2-carboxylate reductase (D.C. 1.5.1.1). The rate of reduction of delta1-piperidine-2-carboxylate was high in certain telencephalic and diencephalic regions, lower in the brain stem, and not measurable in the cerebellum and spinal cord. In addition to delta1-piperidine-2-carboxylate, delta1-pyrroline-2-carboxylate was also found to be a substrate for the same enzyme in homogenates of mouse forebrain. Enzyme kinetic data for both substrates and, in addition, for NADH were derived from determinations using enzyme fractions of mouse telencephalon. The discussion is based on earlier findings concerning the utilisation of D-proline in the neuronal protein synthesis of mouse brain.
