Abstract
Wild type PC12 pheochromocytoma cells express a Na+-dependent norepinephrine transporter that operates in the uptake of catecholamines. In addition to the previously described Na+-dependent system A for the uptake of α-amino-isobutyric acid and system Gly for glycine, we have identified two other Na+-dependent transporter systems for amino acid uptake in these cells: 1) system β for β-alanine and taurine; and 2) a system for creatine. Uptake of α-amino-isobutyric acid, glycine, β-alanine, and creatine is not affected in some PC12 variants that were previously shown to be deficient in catecholamine uptake and to have decreased levels of norepinephrine transporter mRNA. We have isolated two PC12 cDNA clones that are essentially identical in sequence to recently reported cDNAs for rat brain taurine and creatine transporters, respectively, and a third cDNA that appears to code for a novel transporter. mRNAs for these three transporters are present at wild type levels in those variants that express no or little norepinephrine transporter mRNA. These results support the notion that the expression of catecholamine reuptake transporters may be particularly susceptible to down-regulation.
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References
Iversen, L. L. 1970. Neuronal uptake processes for amines and amino acids. Adv. Biochem. Psychopharmacol. 2:109–132.
Christensen, H. N., and Markowske, M. 1985. Recognition chemistry of anionic amino acids for hepatocyte transport and for neurotransmittory action compared. Life Sci. 33:2255–2267.
Shotwell, M. A., Kilberg, M. S., and Oxender, D. L. 1983. The regulation of neutral amino acid transport in mammalian cells. Biochim. Biophys. Acta 737:267–284.
Christensen, H. N. 1984. Organic ion transport during seven decades-the amino acids. Biochim. Biophys. Acta 779:255–269.
Amara, S. G., and Kuhar, M. J. 1993. Neurotransmitter transporters: recent progress. Annu. Rev. Neurosci. 16:73–93.
Guastella, J., Nelson, N., Nelson, H., Czyzyk, L., Keynan, S., Miedel, M. C., Davidson, N., Lester, H. A., and Kanner, B. I. 1990. Cloning and expression of a brain GABA transporter. Science 249:1303–1306.
Giros, B., Mestikawy, S. E., Bertrand, L., and Caron, M. G. 1991. Cloning and functional characterization of a cocaine-sensitive dopamine transporter. FEBS Lett. 295:149–154.
Kilty, J. E., Lorang, D., and Amara, S. G. 1991. Cloning and expression of a cocaine-sensitive rat dopamine transporter. Science 254:578–579.
Pacholcyzk, T., Blakely, R. D., and Amara, S. G. 1991. Expression cloning of a cocaine-and antidepressant-sensitive human noradrenaline transporter. Nature, 350:350–353.
Smith, K. E., Borden, L. A., Hartig, P. R., Branchek, T., and Weinshank, R. L. 1992. Cloning of a glycine transporter reveals colocalization with NMDA receptors. Neuron 8:927–935.
Shimada, S., Kitayama, S., Lin, C.-L., Patel, A., Nanthakumar, E., Gregor, P., Kuhar, M., and Uhl, G. 1991. Cloning and expression of a cocaine-sensitive dopamine transporter complementary DNA. Science, 254:576–578.
Guastella, J., Brecha, N., Weigmann, C., Lester, H. A., and Davidson, N. 1992. Cloning, expression, and localization of a rat brain high-affinity glycine transporter. Proc. Natl. Acad. Sci U. S. A. 89:7189–7193.
Kanai, Y., and Hediger, M. A. 1992. Primary structure and functional characterization of a high affinity glutamate transporter. Nature 360:467–471.
Pines, G., Danbolt, N. C., Bjoras, M., Zhang, Y., Bendahan, A., Eide, L., Koepsell, H., Storm-Mathisen, J., Seeberg, E., and Kanner, B. I. 1992 Cloning and expression of a rat brain L-glutamate transporter. Nature 360:464–467.
Storck, T., Schulte, S., Hofmann, K., and Stoffel, W. 1992. Structure, expression, and functional analysis of a Na+-dependent glutamate/aspartate transporter from rat brain. Proc. Natl. Acad. Sci. U.S.A. 89:10955–10959.
Greene, L. A., and Tischler, A. S. 1976. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl. Acad. Sci. U. S. A. 73:2424–2428.
Greene, L. A., and Rein, G. 1977. Release, storage and uptake of catecholamines by a clonal cell line of nerve growth factor (NGF) responsive pheochromocytoma cells. Brain Res. 129:247–263.
Friedrich, U., and Bonisch, H. 1986. The neuronal noradrenaline transport system of PC12 cells: Kinetic analysis of the interaction between noradrenaline, Na+ and Cl− in transport. Naunyn-Schmeideburgs Arch. Pharmacol. 333:246–252.
Koide, M., Cho, A. K., and Howard, B. D. 1986. Characterization of xylamine binding to proteins of PC12 pheochromocytoma cells. J. Neurochem. 47:1277–1285.
Ramachandran, B., Houben, K., Rozenberg, Y. Y., Haigh, J. R., Varpetian, A., and Howard, B. D. 1993. Differential expression of transporters for norepinephrine and glutamate, in wild type, variant, and Wnt1-expressing PC12 cells. J. Biol. Chem. 268:23891–23897.
Bitler, C. M., Zhang, M.-B., and Howard, B. D. 1986. PC12 variants deficient in catecholamine transport. J. Neurochem. 47:1286–1293.
Howard, B. D., Cho, A. K., Zhang, M., Koide, M., and Lin, S. 1990. Covalent labeling of the cocaine-sensitive catecholamine transporter. J. Neurosci. Res. 26:149–158.
Denton, T., and Howard, B. D. 1987. A dopaminergic cell line variant resistant to the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydro-pyridine. J. Neurochem. 49:622–630.
Andersen, J. K., Zhang, M.-b., Zhong, X.-h., Rozenberg, Y. Y., and Howard, B. D. 1990. MPTP-Resistant flat cell PC12 variants having a partial loss of transformed phenotype. J. Neurochem. 55:559–567.
Seed, B., and Aruffa, A. 1987: Molecular cloning of the CD2 antigen, the T-cell erythrocyte receptor, by a rapid immunoselective procedure. Proc. Natl. Acad. Sci. U.S.A. 84:3365–3369.
Liu, Y., Roghani, A., and Edwards, R. H. 1992. Gene transfer of a reserpine-sensitive mechanism of resistance to N-methyl-4-phenylpyridinium. Proc. Natl. Acad. Sci. U. S. A. 89:9074–9078.
Barnes, W. M. 1987. Sequencing DNA with dideoxyribonucleotides as chain terminators: Hints and strategies for big projects. Methods Enzymol. 152:538–558.
Chirgwin, J. M., Przbyla, A. E., MacDonald, R. J., and Rutter, W. J. 1979. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299.
Aviv, H., and Leder, P. 1972. Purification of biologically active globin messenger RNA by chromatography on oligothymidilic acidcellulose. Proc. Natl. Acad. Sci. U.S.A. 69:1408–1412.
Maniatis, T., Fritsch, E. F., and Sambook, J., 1982. Molecular Cloning, pp. 10545, Cold Spring Harbor Laboratory, New York.
Thomas, P. 1980. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. U.S.A. 77:5201–5205.
Stanley, P. E., and Williams, S. G. 1962. Use of the scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Anal. Biochem. 29:381–392.
Strehler, B. L., and Totter, J. K. 1954. Determination of ATP and related compounds: firefly luminescence and other methods. Methods Biochem. Anal. 1:341–346.
McGuire, J. C., and Greene, L. A. 1979. Rapid stimulation by nerve growth factor of amino acid uptake by clonal PC12 pheochromocytoma cells. J. Biol. Chem. 254:3362–3367.
Moller, A., and Hamprecht, B. 1989. Creatine transport in cultured cells of rat and mouse brain. J. Neurochem. 52:544–550.
Tierney, N. A. and Peters, J. P. 1943. The mode of excretion of creatine and creatine metabolism in thyroid disease. J. Clinic. Invest. 22:595–602.
Allinson, M. J. C. 1945. A specific enzymatic method for the determination of creatine and creatinine in blood. J. Biol. Chem. 157:169–172.
Wallimann, T., Wyss, M., Brdiczka, D., Nicolay, K., and Eppenberger, H. M. 1992. Intracellular compartmentation, structure and function of creatine isoenzymes in tissues with high and fluctuating energy demands: the “phosphocreatine circuit” for cellular energy homeostasis. Biochem. J. 281:21–40.
Reynolds, E. E., Melega, W., and Howard, B. D. 1982. Adenosine 5′-triphosphate independent secretion from PC12 pheochromocytoma cells Biochemistry 21:4795–4799.
Smith, K. E., Borden, L. A., Wang, C. D., Hartig, P. R.., Branchek, T., and Weinshank, R. L. 1992. Cloning and expression of a high affinity taurine transporter from rat brain. Mol. Pharmacol. 42:563–569.
Guimbal, C., and Kilimann, M. W. 1993: A Na+-dependent creastine transporter in rabbit brain, muscle, heart, and kidney. J. Biol. Chem. 268:8418–8421.
Liu, Q.-R., Mandiyan, S., Lopez-Corcuera, B., Nelson, H., and Nelson, N. 1993. A rat brain cDNA encoding the neurotransmitter transporter with an unusual structure. FEBS Lett. 315:114–118.
Fitch, C. D., Shields, R. P., Payne, W. F., and Dacus, J. M. 1968. Creatine metabolism in skeletal muscle. J. Biol. Chem. 243:2024–2027.
Kong, C.-T., Yet, S.-F., and Lever, J. E. 1993. Cloning and expression of a mammalian Na+/amino acid cotransporter with sequence similarity to Na+/glucose cotransporters. J. Biol. Chem. 268:1509–1512.
Bannon, M. J., Poosch, M. S., Xia, Y., Goebel, D. J., Cassin, B., and Kapatos, G. 1992. Dopamine transporter mRNA content in human substantia nigra decreases precipitously with age. Proc. Natl. Acad. Sci. U.S.A. 89:7095–7099.
Xia, Y., Goebel, D. O., Kapatos, G., and Bannon, M. J. 1992. Quantitation of rat dopamine transporter m RNA: Effects of cocaine treatment and withdrawal. J. Neurochem. 59:1179–1182.
Uhl, G. R., Kitayama, S., Gregor, P., Nanthaakumar, E., Perisco, A., and Shimada, S. 1992. Neurotransmitter transporter family cDNAs in a rat midbrain library: orphan transporters suggest sizable structural variations. Mol. Brain Res. 16:353–359.
Mayser, W., Schloss, P., and Betz, H. 1992. Primary structure and functional expression of a choline transporter, expressed in the rat nervous system. FEBS Lett. 305:31–36.
Melega, W. P., and Howard, B. D. 1980. Choline and acetylcholine metabolism in PC12 secretory cells. Biochemistry 20:4477–4483.
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Houben, K., Dardashti, K. & Howard, B.D. PC12 variants deficient in norepinephrine transporter mRNA have wild type activities of several other related transporters. Neurochem Res 19, 743–751 (1994). https://doi.org/10.1007/BF00967715
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DOI: https://doi.org/10.1007/BF00967715