The polyamines exhibit a number of neurophysiological and metabolic effects related to control of normal development and maintenance of central nervous system (CNS) homeostasis. These include control of cell growth and differentiation, regulation of nucleic acid and protein synthesis, modulation of ionic channels and calcium-dependent transmitter release. In the developing neuron, polyamines have been associated with programmed cell death, neurodegeneration and with neurotrophic functions. Disruption of these processes, due to inherited defects of polyamine metabolism, is therefore certain to have neuro-pathophysiological consequences most likely leading to anomalous development and neurodegeneration. Our long-term goal is to study neurochemical, physiological and pathological consequences of inherited defects of polyamine metabolism in the human. Such studies will lead to definition of pathophysiology, which in turn will help delineate the normal roles of polyamines within the CNS. To attain our goal we need a mechanism for identification of patients with inborn errors of polyamine metabolism. The short-term specific aims are therefore: 1) to establish and validate HPLC methodology that allows accurate measurement of polyamines and metabolites in cerebrospinal fluid. Using precolumn derivatization with N-hydroxysuccinimidyl-6-aminoquinoylcarbamate, ternary gradient separation and fluorescence detection, we will establish and validate a method to allow measurement of putrescine, spermidine, spermine and the acetyl derivatives of spermine and spermidine in cerebrospinal fluid (CSF). This method will facilitate the search for inherited defects that affect this area of metabolism. 2) To determine age related normal control and disease control reference ranges, to examine the effect of gender and to establish the presence or absence of a rostro-caudal gradient for polyamines in CSF Defining these variables is vital if meaningful interpretation of patient results is to be made. 3) To search for polyamine defects in infants and children with neurological disease of unknown origin. We will analyze over 1000 CSF samples collected retrospectively and over 1000 samples collected prospectively. Clinical data will be available. All clinical and laboratory data will be entered into a Microsoft Access Database to permit analysis across the entire sample, within subsets and between subsets. Discovery of humans with inherited defects affecting polyamine metabolism, with description of metabolic and clinical phenotype and with possible neuropathology, will certainly provide clues as to pathophysiological mechanisms, will further help to elucidate the major physiological functions of the polyamines, and may allow differentiation of the individual roles of putrescine, spermidine and spermine depending on the exact site of the metabolic block.
