Skip to main content
Log in

Utilization of alpha-ketoglutarate as a precursor for transmitter glutamate in cultured cerebellar granule cells

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Alpha-ketoglutarate together with an amino group donor (alanine) was shown to be able to serve as a precursor for the glutamate pool which is released by potassium-induced depolarization (i.e., transmitter glutamate) in cerebellar granule cells. However, these compounds could not be utilized as precursors for intracellular glutamate or for release of transmitter aspartate. The formation of transmitter glutamate was inhibited by the transamination inhibitor aminooxyacetic acid but not by phenylsuccinate, an inhibitor of the dicarboxylate carrier in the mitochondrial membrane. Both of these inhibitors have previously been found to inhibit synthesis of transmitter glutamate from glutamine. The results support the hypothesis that alpha-ketoglutarate and alanine undergo transamination in the cytosol to form pyruvate and glutamate, and that this glutamate pool is available for transmitter release of glutamate but does not constitute the major intracellular pool of glutamate.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Hertz, L. 1979. Functional interactions between neurons and astrocytes. I. Turnover and metabolism of putative amino acid transmitters. Progr. Neurobiol. 13:277–323.

    Google Scholar 

  2. Hertz, L., and Schousboe, A. 1986. Role of astrocytes in compartmentation of amino acid and energy metabolism. Pages 179–208,in Fedoroff, S., and Vernadakis, A. (eds.), Astrocytes, Vol. 2, Academic Press, New York.

    Google Scholar 

  3. Hertz, L., and Schousboe, A. 1988. Metabolism of glutamate and glutamine in neurons and astrocytes in primary cultures. Pages 39–55,in Kvamme, E. (ed.), Glutamine and Glutamate in Mammals, Fl., Vol. 2. CRC Press, Boca, Raton.

    Google Scholar 

  4. Shank, R. P., and Aprison, M. H., Glutamate as a neurotransmitter. 1988. Pages 3–19,in Kvamme, E. (ed.), Glutamine and Glutamate in Mammals, Fl. Vol. 2, CRC Press, Boca Raton.

    Google Scholar 

  5. Shank, R. P., and Campbell, G. LeM. 1981. Avid Na+-dependent, high affinity uptake of alpha-ketoglutarate by nerve terminal enriched material from mouse cerebellum. Life Sci. 28:843–850.

    Google Scholar 

  6. Shank, R. P., and Campbell, G. L. 1982. Glutamine and alphaketoglutarate uptake and metabolism by nerve terminals enriched material from mouse cerebellum. Neurochem. Res. 7:601–616.

    Google Scholar 

  7. Shank, R. P. and Campbell, G. 1984. Alpha-ketoglutarate and malate uptake and metabolism by synaptosomes. Further evidence for an astrocyte to neuron metabolic shuttle. J. Neurochem., 42:1153–1161.

    Google Scholar 

  8. Shank, R. P., Baldy, W. J., and Ash, C. W. 1989. Glutamine and 2-oxoglutarate as metabolic precursors of the transmitter pools of glutamate and GABA: Correlation of regional uptake by rat brain synaptosomes. Neurochem. Res. 14:371–376.

    Google Scholar 

  9. Hertz, L., Schousboe, A., Boechler, N., Mukerji, S., and Federoff, S. 1978. Kinetic characteristics of glutamate uptake into normal astrocytes in cultures. Neurochem. Res. 3:1–14.

    Google Scholar 

  10. McLennan, H. 1976. The autoradiographic localization of L-[3H]-glutamate in rat brain tissue. Brain Res., 115:139–144.

    Google Scholar 

  11. Schousboe, A., Drejer, J., and Hertz, L. 1988. Uptake and release of glutamate and glutamine in neurons and astrocytes in primary cultures. Pages 21–38, in Kvamme, E. (ed.), Glutamine and Glutamate in Mammals, Fl. Vol. 2, CRC Press, Boca Raton.

    Google Scholar 

  12. Yu, A. C. H., Schousboe, A., and Hertz, L. 1982. Metabolic fate of (14C)-labelled glutamate in astrocytes. J. Neurochem., 39:954–960.

    Google Scholar 

  13. Waniewski, R. A., and Martin, D. L. 1986. Exogenous glutamate is metabolized to glutamine and exported by rat primary astrocyte cultures. J. Neurochem., 47:304–313.

    Google Scholar 

  14. Hertz, L., Yu, A., Svenneby, G., Kvamme, E., Fosmark, H., and Schousboe, A. 1980. Absence of preferential uptake of glutamine into neurons—an indication of a net transfer of TCA constituents from nerve endings to astrocytes? Neurosci. Lett., 16:103–109.

    Google Scholar 

  15. Shank, R. P., Bennett, G. S., Freytag, S. O., and Campbell, G. LeM. 1985. Pyruvate carboxylase: an astrocyte-specific enzyme implicated in the replenishment of amino acid neurotransmitter pools. Brain Res. 329:364–367.

    Google Scholar 

  16. Yu, A. C. H., Drejer, J., Hertz, L., and Schousboe, A. 1983. Pyruvate carboxylaste activity in primary cultures of astrocytes and neurons. J. Neurochem. 41:1484–1487.

    Google Scholar 

  17. Kaufman, E. E. 1990. The effect of [K+] on CO2 fixation in cultured glial cells, Trans. Amer. Soc. Neurochem. 21:289.

    Google Scholar 

  18. Peng, L., Schousboe, A., and Hertz, L. 1989. Utilization of alpha-ketoglutarate as a precursor of transmitter glutamate. Trans. Amer. Soc. Neurochem. 20:266.

    Google Scholar 

  19. Schousboe, A., Meier, E., Drejer, J. and Hertz, L. 1989. Preparation of primary cultures of mouse (rat) cerebellar granule cells. Pages 203–206,in Shahar, A, de Vellis, J, Vernadakis, A. and Haber, B. (eds.) A Dissection and Tissue Culture Manual for the Nervous System. Alan R. Liss, New York.

    Google Scholar 

  20. Hertz, L., Juurlink, B. H. J., Fosmark, H., and Schousboe, A. 1982. Astrocytes in primary culture. Pages 175–186,in Pfeiffer, S. E. (ed.), Neuroscience Approached Through Cell Culture, Vol. 1, CRC Press, Boca Raton, FL.

    Google Scholar 

  21. Peng, L., Juurlink, B. H. J., and Hertz, L. Development of cerebellar granule cells in the presence and absence of excess extracellular potassium-do the two culture systems provide means of distinguishing between events in transmitter-related and non-transmitter-related glutamate pools? (submitted for publication).

  22. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193:265–275.

    Google Scholar 

  23. Lindroth, P., and Mopper, R. 1979. High performance liquid chromatographic determination of subpicomole amounts of amino acids by precolumn fluorescence derivatization with O-phthaldialdehyde. Anal. Chem., 51:1667–1674.

    Google Scholar 

  24. Yu, A. C. H., Hertz, E., and Hertz, L. 1984. Alterations in uptake and release rates for GABA, glutamate and glutamine during biochemical maturation of highly purified cultures of cerebral cortical neurons, a GABAergic preparation. J. Neurochem. 42:952–960.

    Google Scholar 

  25. Kihara, M., and Kubo, T. 1988. Aspartate aminotransferase for synthesis of transmitter glutamate in the medulla oblongata: Effect of aminoxyacetic acid and 2-oxyoglutarate. J. Neurochem. 52:1127–1134.

    Google Scholar 

  26. Hertz, L., Peng, L., Hertz, E., Juurlink, B. H. J., and Yu, P. H. 1989. Development of monoamine oxidase activity and monoamine effects on glutamate release in cerebellar neurons and astrocytes. Neurochem. Res. 14:1039–1046.

    Google Scholar 

  27. Palaiologos, G., Hertz, L., and Schousboe, A. 1988. Evidence that aspartate aminotransferase activity and ketodicarboxylate carrier function are essential for biosynthesis of transmitter glutamate. J. Neurochem., 51:317–320.

    Google Scholar 

  28. Palaiologos, G., Hertz, L., and Schousboe, A. 1989. Role of aspartate aminotransferase and mitochondrial dicarboxylate transport for release of endogenously and exogenously supplied neurotransmitter in glutamatergic neurons. Neurochem. Res., 14:359–366.

    Google Scholar 

  29. Kvamme, E. 1983. Glutaminase (PAG). Pages 51–67,in Hertz, L, Kvamme, E, McGeer, E. G., and Schousboe, A. (eds.) Glutamine, Glutamate and GABA in the Central Nervous System, Alan R. Liss, New York.

    Google Scholar 

  30. Yudkoff, M., Nissim, I., Hummeler, K., Medow, M., and Pleasure, D. 1986. Utilization of [15N] glutamate by cultured astrocytes. Biochem. J. 234:185–192.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Peng, L., Schousboe, A. & Hertz, L. Utilization of alpha-ketoglutarate as a precursor for transmitter glutamate in cultured cerebellar granule cells. Neurochem Res 16, 29–34 (1991). https://doi.org/10.1007/BF00965824

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00965824

Key words

Navigation