Abstract
Buffered proline was injected subcutaneously into rats twice a day at 8 h intervals from the 6th to the 28th day of age. Control rats received saline in the same volumes. The animals were weighed and killed by decapitation 12 h after the last injection. Cerebral cortex was used for the determination of Na+,K+-ATPase and Mg2+-ATPase activities. Body, whole brain and cortical weights were similar in the two groups. Na+,K+-ATPase activity was significantly reduced (by 20%) in membranes from the proline-treated group compared to the controls, whereas Mg2+-ATPase activity was not affected by proline. In another set of experiments, synaptic plasma membranes were prepared from cerebral cortex of 29-day-old rats and incubated with proline at final concentrations ranging from 0.1 to 2.0 mM. Na+,K+-ATPase activity, but not Mg2+-ATPase activity, was inhibited by 20-30%. Since proline concentrations in plasma of chronically treated rats and of type ll hyperprolinemic children are of the same order of magnitude as those tested in vitro, the results suggest that reduction of Na+,K+-ATPase activity may contribute to the neurological dysfunction found in some patients affected by type ll hyperprolinemia.
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REFERENCES
Ault, B., Wang, C.M., and Yawn, B.C. (1987). L-Proline depolarizes rat spinal motoneurones by an excitatory amino acid-sensitive mechanism. Br. J. Pharmacol. 92:319–326.
Ben-Ari, Y. (1985). Limbic seizure and brain damage produced by kainic acid: mechanisms and relevance to human temporal lobe epilepsy. Neuroscience, 14: 375–403.
Bertorello, A.M., and Kats, A.L. (1995). Regulation of Na+-K+-Pump activity: pathways between receptors and effectors. NIPS, 10:253–259.
Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-die binding. Anal. Biochem. 72:248–254.
Chan, K.M., Delfert, D., Junger, K.D. (1986). A direct colorimetric assay for Ca+2-stimulated ATPase activity. Anal. Biochem. 157:375–380.
Cherkin, A., Eckardt, M.J., and Gerbrandt, L.K. (1976). Memory: proline induces retrograde amnesia in chicks. Science 193:242–244.
Choi, D.W. and Rothman, S.M. (1990). The role of glutamate neurotoxicity in hypoxic-ischemic neuronal death. Ann. Rev. Neurosci. 13:171–182.
Cohen, M.S.and Nadler, J.V. (1997). Proline-induced potentiation of glutamate transmission. Brain Res. 761:271–282.
Cousin, M.A., Nicholls, D.G. and Pocock, J.M. (1995). Modulation of ion gradients and glutamate release in cultured cerebellar granule cells by ouabain. J. Neurochem. 64:2097–2104.
Ericinska, M. and Silver, I.A. (1994). Ions and energy in mammalian brain. Neurobiol. 43:37–71.
Flynn, M.P., Martin, M.C., Moore, P.T., Stafford, J.A., Fleming, G.A., and Phang, J.M. (1989). Type II hyperprolinaemia in a pedigree of Irish travellers (normands). Arch. Dis. Child. 64: 1699–1707.
Fremeau, R.T., Caron, M.G., and Blakely, R.D. (1992). Molecular cloning and expression of a high affinity L-proline transporter expressed in putative glutamatergic pathways of rat brain. Neuron 8:915–926.
Fremeau, R.T., Velaz-Faircloth, M., Miller, J.W., Henzi, V.A., Cohen, S.M., Nadler, J.V., Shafqat, S., Blakely, R.D., and Domin, B. (1996). A novel nonopioid action of enkephalins: competitive inhibition of the mammalian brain high affinity L-proline transporter. Mol. Pharmacol. 49:10333–1041.
Henzi, V., Reichiling, D.B., Helm, S.W., and MacDermott, A.B. (1992). L-proline activates glutamate and glycine receptors in culture rat dorsal horn neurons. Mol. Pharmacol. 41:793–801.
Jones, D.H and Matus, A.I. (1974). Isolation of synaptic plasma membrane from brain by combination flotation-sedimentation density gradient centrifugation. Biochim. Biophys. Acta 356: 276–287.
Lees, G.J., Lehmann, A., Sandberg, M., and Hamberg, H. (1990). The neurotoxicity of ouabain, a sodium-potassium ATPase inhibitor, in the rat hippocampus. Neurosci. Lett. 120:159–162.
Lees, G.J. (1993). Contributory mechanisms in the causation of neurodegenative disorders. Neuroscience 54:287–322.
Lees, G.J. and Leong, W. (1995). Brain lesions induced by specific and non-specific inhibitors of sodium-potassium ATPase. Brain Res. 649:225–233.
Martin, D., Ault, B., and Nadler, J.V. (1992). NMDA receptor-mediated depolarizing action of proline on CA1 pyramidal cells. Eur. J. Pharmacol. 219: 59–66.
Moreira, J.C.F., Wannmacher, C.M.D., Costa, S.M., and Wajner, M. (1989). Effect of proline administration on rat behavior in aversive and nonaversive tasks. Pharmac. Biochem. & Behav. 32: 885–890.
Nadler, J.V. and Cohen, S.M. (1995). Proline inhibits the release of glutamate and aspartate from Schaffer collateral-commissural terminal in rats. Soc. Neurosci. Abstr. 21: 353.
Nadler, J.V., Bray, S.D., and Evenson, D.A. (1992). Autoradiographic localization of proline uptake in excitatory hippocampal pathways. hippocampus 2:269–278.
Nistri, A. and Morelli, P. (1978). Effects of proline and other neutral amino acids on ventral root potentials of the frog spinal cord in vitro. Neuropharmacology 17: 21–27.
Oton, D.S. and Markowska, L. (1994). Memory and hippocampal function as targets for neurotoxic substances. Neurotoxicology 15: 439–444.
Pace, J.R., Martin, B.M., Paul, S.M., and Rowski, A. (1992). High concentration of neutral amino acids activate NMDA receptor currents in rat hippocampal neurons. Neurosci. Lett. 141: 97–100.
Pavone, L., Mollica, F. and Levy, H.L. (1975). Asymptomatic type-11 hypeprolinemia associated with hyperglicinaemia in three sibs. Arch. Dis. Child., 50: 637.
Phang, J.M., Yet, G.C. and Scriver, C.R (1995). Disorders of proline and hydroxyproline metabolism. In: Scriver, C.R., Beaudet, A.L., Sly, W.S. and Valle, D. eds. The Metabolic and Molecular Bases of Inherited Disease, McGraw-Hill, Inc., New York, 7th Ed., pp. 1015–1075.
Renkawek, K., Renier, W.O., De Pont, J.J.H.H.M., Vogels, O.J.M., and Gabreels, F.J.M. (1992). Neonatal status convulsivus, spongiform encephalopathy, and low activity of Na+/K+-ATPase in the brain. Epilepsy 33: 58–64.
Rohads, D.E., Peterson, N.A., and Raghupathy, E. (1983). Selective inhibition of synaptosomal proline uptake by leucine and methionine enkephalins. J. Biol. Chem. 258: 12233.
Satoh, E. and Nakazato, Y. (1992). On the mechanism of ouabain-induced release of acetylcholine from synaptosomes. J. Neurochem. 58:1038–1044.
Shafqat, S., Velaz-Faircloth, M., Henzi, V.A., Witney, K.D., Yang-Feng, T.L., Seldin, M.F., and Fremeau, R.T. (1995). Human brain-specific L-proline transporter: molecular cloning, functional expression, and chromosomal localization of the gene in human and mouse genomes. Mol. Pharmacol. 48: 219–229.
Tsakiris, S. and Deliconstantinos, G. (1984). Influence of phosphatidylserine on (Na+ + K+)-stimulated ATPase and acetylcholinesterase activities of dog brain synaptosomal plasma membranes. Biochem. J. 22: 301–307.
Van Herreveld, A. and Fifkova, E. (1973). Effects of amino acids on the isolated chicken retina, and on its response to glutamate stimulation. J. Neurochem., 20: 947.
Velaz-Faircloth, M., Guadano-Ferraz, A., Henzi, V.A., and Fremeau, R.T. (1995). Mammalian brain-specific L-proline transporter. Neuronal localization of mRNA and enrichment of transporter protein in synaptic plasma membranes. J. Biol. Chem. 270:15755–15761.
Wyse, A.T.S., Brusque, A.M., Silva, C.G., Streck, E.L., Wajner, M., and Wannmacher, C.M.D. (1998). Inhibition of Na+, K+-ATPase from rat brain cortex of rats by propionic acids. NeuroReport, 9:1719–1721.
Wyse, A.T.S., Noriler, M.E., Borges, L.F., Floriano, P.J., Silva, C.G., Wajner, M. and Wannmacher, C.M.D. (1999). Alanine prevents the decrease of Na+, K+-ATPase activity in experimental phenylketonuria. Metab. Brain Dis. 14: 95–101.
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Pontes, Z.E., Oliveira, L.S., Baveresco, C.S. et al. Proline Administration Decreases Na+,K+-ATPase Activity in the Synaptic Plasma Membrane from Cerebral Cortex of Rats. Metab Brain Dis 14, 265–272 (1999). https://doi.org/10.1023/A:1020789109913
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DOI: https://doi.org/10.1023/A:1020789109913