Skip to main content
Log in

Identification and characterization of the beta-adrenergic receptor on neuroblastoma × glioma hybrid NG108-15 cells

  • Published:
Cellular and Molecular Neurobiology Aims and scope Submit manuscript

Summary

  1. 1.

    Using [3H]DHA and unlabeledl-alprenolol, a substantial amount of over 64% specific binding ofβ-adrenergic receptor has been identified on the neuroblastoma × glioma hybrid NG108-15 cell, which has been proven to display numerous functional characteristics of intact neurons.

  2. 2.

    Beta-adrenergic receptor binding on intact NG108-15 cells does not change significantly upon morphological differentiation, induced by 1 mM dibutyryl cyclic AMP (dBcAMP).

  3. 3.

    The [3H]DHA binding on intact NG108-15 cells is rapid, saturable, and reversible, having at 1/2 of 1.0 min for association and 3.5 min for dissociation.

  4. 4.

    The affinity constant (K d) and maximum binding capacity (B max) for binding of [3H]DHA toβ-adrenergic receptors on NG108-15 cells have been estimated by Scatchard plot analysis to be 2.5 and 0.23 nM, respectively. Further analysis indicates a single class of receptors for [3HDHA binding on NG108-15 cells.

  5. 5.

    Studies on kinetic properties have revealed on-rate (K + 1) and off-rate (K - 1) constants of 0.7 × 10−9 M min−1 and 0.19 min−1, respectively. Further, the IC50 value and inhibition constant (K i) for unlabeledl-alprenolol to inhibit [3HDHA binding on NG108-15 cells have been estimated to be 10−5 and 8.9 × 10−6 M, respectively.

  6. 6.

    The rank-order potency of catecholamine agonists, (−)ISO > (+)ISO > EPI > NE, reveals the presence of type 2 receptor for theβ-adrenergic binding on both differentiated and undifferentiated NG108-15 cells.

  7. 7.

    The present study indicates that the clonal neuroblastoma × glioma hydbrid NG108-15 cell line possesses substantial amounts ofβ-adrenergic receptors with characteristics similar to those on neuronal cells.

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

  • Alexander, R. W., Davis, J. N., and Lefkowitz, R. J. (1975). Direct indentification and characterization of beta-adrenergic receptors in rat brain.Nature 258437–440.

    Google Scholar 

  • Cheng, Y. C., and Prusoff, W. H. (1973). Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50% inhibition (IC50) of an enzymatic reaction.Biochem. Pharmacol. 223099–3108.

    Google Scholar 

  • Galand, S. P., and Allred, S. (1981). Binding and functional characteristics of beta-adrenergic receptors in the intact neutrophil.J. Lab. Clin. Med. 98227–237.

    Google Scholar 

  • Ghahary, A., and Cheng, K. W. (1989). Characterization of muscarinic acetylcholine receptors on intact neuroblastoma × glioma NG108-15 cells upon induced differentiation.Mol. Cell. Biochem. 86181–188.

    Google Scholar 

  • Ghahary, A., Vriend, J., and Cheng, K. W. (1989). Modification of the indolamine content in neuroblastoma × glioma hybrid NG108-15 cells upon induced differentiation.Cell. Mol. Neurobiol. 9343–355.

    Google Scholar 

  • Hamprecht, B. (1977). Structure, electrophysiological, biochemical and pharamacological properties of neuroblastoma × glioma hybrids in cell culture.Int. Rev. Cytol. 4990–170.

    Google Scholar 

  • Hamprecht, B., and Schultz, J. (1973). Influence of noradrenalin, prostaglandin El and inhibitors of phophodiesterase activity on the levels of the cyclic adenosine 3,5,-monophosphate in somatic cells hybrids.Physiol. Chem. 3541635–1641.

    Google Scholar 

  • Hill, A. V. (1910). The possible effects of the aggregation of the molecules of hemoglobin on its dissociation curve.J. Physiol. 40iv-vii.

    Google Scholar 

  • Insel, P. A., and Stoolman, L. M. (1978). Radioligand binding to beta adrenergic receptors of intact cultured S49 cells.Mol. Pharmacol. 14549–561.

    Google Scholar 

  • Insel, P. A., Mahan, L. C., Motulsky, H. V., Stoolman, L. M., and Koachman, A. M. (1983). Time-dependent decreases in binding affinity of agonists forβ-adrenergic receptors of intact S49 lymphoma cells.J. Biol. Chem. 25813597–13605.

    Google Scholar 

  • Kahn, D. J., Mitrius, J. C., and U'Prichard, D. C. (1981). Alpha-adrenergic receptors in neuroblastoma × glioma hybrid cells: Characterization with agonist and antagonist radioligands and relationship to adenylate cyclase.Mol. Pharmacol. 2117–26.

    Google Scholar 

  • Klee, W., and Nirenberg, M. (1974). Neuroblastoma × glioma hybrid cell line with morphine receptors.Proc. Natl. Acad. Sci. USA 713474–3477.

    Google Scholar 

  • Lands, A. M., Arnold, A., McAuliff, J. P., Luduena, F. P., and Brown, J. G., Jr. (1967). Differentiation of receptor systems activated by sympathomimetic amines.Nature 214597–598.

    Google Scholar 

  • Maderspach, L., and Fajszi, C. (1982).β-Adrenergic receptors of brain cell membrane, intergrity implies apparent positive cooperativity and high affinity.Biochim. Biophys. Acta. 692469–478.

    Google Scholar 

  • McGee, R., Simpson, P., Christian, C., Mata, M., Nelson, P., and Nirenberg, M. (1978). Regulation of acetylcholine release from neuroblastoma × glioma hybrid cells.Proc. Natl. Acad. Sci. USA 751314–1318.

    Google Scholar 

  • McPherson, G. A. (1983). A pratical computer-based approach to the analysis of radioligand binding experiments.Comp. Prog. Biomed. 17107–113.

    Google Scholar 

  • Motulsky, H. V., and Insel, P. A. (1982). Adrenergic receptors in man: Direct indentification, physiologic regulation, and chemical alterations.N. Engl. J. Med. 307 18–29.

    Google Scholar 

  • Nelson, P. G., Christian, C. N., Daniels, M. P., Henkart, M., Bullock, P., Mullinax, D., and Nirenberg, M. (1978). Formation of synapses between cells of a neuroblastoma × glioma clone and mouse myotubes.Brain Res. 147245–259.

    Google Scholar 

  • Porzig, H. (1982). Are there differences in theβ-adrenergic-adenylate cyclase systems of fragmented membranes and living cells?Trends Pharamol. Sci. 375–78.

    Google Scholar 

  • Repke, H, and Maderspach, L. (1983). Muscarinic acetylcholine receptors on cultured glial cells.Brain Res. 232206–211.

    Google Scholar 

  • Rosenthal, H. (1967). Graphic method for the determination and presentation of binding parameters in a complex system.Anal. Biochem. 20525–532.

    Google Scholar 

  • Sanders, R. B., Thompson, W. J., and Robison, G. A. (1977). Epinephrine and glucagon-stimulated cardiac adenylate cyclase activity.Biochim. Biophys. Acta 49810–20.

    Google Scholar 

  • Scatchard, G. (1949). The attraction of proteins for small molecules and ions.Ann. N.Y. Acad. Sci. 51660–672.

    Google Scholar 

  • Strange, P. G., Birdsall, N. J. M., and Burgen, A. S. V. (1978). Ligand binding properties of muscarinic acetylcholine receptor in mouse neuroblastoma cells.Biochem. J. 172495–501.

    Google Scholar 

  • Traber, J., Fischer, K., Buchen, C., and Hamprecht, B. (1975). Muscarinic response to acetylcholine in neuroblastoma × glioma hybrid cells.Nature 255558–560.

    Google Scholar 

  • Whitesett, J. A., Noguchi, A., Neely, J. E., John, C. L., and Moore, J. J. (1981). Beta l-adrenergic receptors in human neuroblastoma.Brain Res. 21673–87.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ghahary, A., Cheng, K.W. Identification and characterization of the beta-adrenergic receptor on neuroblastoma × glioma hybrid NG108-15 cells. Cell Mol Neurobiol 10, 337–350 (1990). https://doi.org/10.1007/BF00711179

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00711179

Key words

Navigation