Skip to main content
Log in

Receptors for phorbol esters are primarily localized in neurons: Comparison of neuronal and glial cultures

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

Abstract

Binding of [3H]PDB has been measured in the present study to determine the levels of protein kinase C in the neuronal and astrocytic glial cells in culture from rat brain. Binding of [3H]PDB to homogenates of cultured neuronal cells from the brains of normotensive and hypertensive rats was time-dependent and specific. The relative potency for competition by various phorbol esters to [3H]PDB binding was TPA > β-PDD > POE > α-PDD ≥4αphorbol. Scatchard analysis showed that neuronal cultures from normotensive rat brains contained 2–3 fold more phorbol ester receptors compared with the glial cultures from the same brains. No differences in theK d andB max were observed between neuronal cultures from normotensive and spontaneously hypertensive rat brains. These studies suggest that the phorbol ester receptors are primarily localized in 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

  1. Kuo, J. F., Andersson, R. G. G., Wise, B. C., Mackerlova, L., Salomonsson, I., Brackett, N. C., Katoh, N., Shoji, M., and Wrenn, R. W. 1980. Calcium-dependent protein kinase: Widespread occurrence in various tissues and phyla of the animal kingdom and comparison of effects of phospholipid, calmodulin, and trifluoparazine. Proc. Natl. Acad. Sci., U.S.A. 77:7039–7043.

    Google Scholar 

  2. Girard, P. R., Mazzei, G. J., Wood, J. G., and Kuo, J. F. 1985. Polyclonal antibodies to phospholipid/Ca2+-dependent protein kinase and immunocytochemical localization of the enzyme in rat brain. Proc. Natl. Acad. Sci. U.S.A., 82:3030–3034.

    PubMed  Google Scholar 

  3. Gonzales, R. A., Feldstein, J. B., Crews, F. T., and Raizada, M. K. 1985. Receptor-mediated inositide hydrolysis is a neuronal response: comparison of primary neuronal and glial cultures. Brain Res 345:350–355.

    PubMed  Google Scholar 

  4. Gonzales, R. A., Crews, F. T., Sumners, C., and Raizada, M. K. Norepinephrine mediated uncoupling of alpha1-adrenergic receptors from phosphoinositide hydrolysis in primary neuronal cultures is rapid and separate from downregulation of the receptors. J. Pharm. Exp. Ther. (in press).

  5. Berridge, M. J. 1984. Inositol triphosphate and diaglycerol as second messengers. Biochem. J., 220:345–360.

    PubMed  Google Scholar 

  6. Castagna, M., Takai, Y., Kaibuchi, K., Sano, K., Kikkawa, U., and Nishizuka, Y. 1982. Direct activation of calciumactivated, phospholipid-dependent protein kinase by tumorpromoting phorbol esters. J. Biol. Chem. 257:7847–7851.

    PubMed  Google Scholar 

  7. Niedel, J. E., Kuhn, L. J., and Vanderbark, G. R. 1983. Phorbol diester receptor co-purifies with protein kinase C. Proc. Natl. Acad. Sci. U.S.A. 80:36–40.

    PubMed  Google Scholar 

  8. Raizada, M. K. 1983. Localization of insulin-like immunoreactivity in the neurons from primary cultures of rat brain. Exp. Cell. Res. 143:351–357.

    PubMed  Google Scholar 

  9. Raizada, M. K., Muther, T. F., and Sumners, C. 1984. Increased angiotensin II specific receptors in neuronal cultures of hypertensive rat brain Am. J. Physiol. 247:C364-C372.

    PubMed  Google Scholar 

  10. Sumners, C., Phillips, M. I., and Raizada, M. K. 1983. Rat brain cells in primary culture: visualization and measurement of catecholamines. Brain Res. 264:267–275.

    PubMed  Google Scholar 

  11. Sumners, C., Muther, T. F., and Raizada, M. K. 1985. Altered norepinephrine uptake in neuronal cultures from spontaneously hypertensive rat brain. Am. J. Physiol. 248:C488-C497.

    PubMed  Google Scholar 

  12. Clarke, D. W., Boyd, F. T., Kappy, M. S., and Raizada, M. K. 1984. Insulin binds to specific receptors and stimulates 2-deoxy-D-glucose uptake in cultured glial cells from rat brain. J. Biol. Chem. 259:11672–11675.

    PubMed  Google Scholar 

  13. Scatchard, G. 1949. The attraction of protein for small molecules and ions. Ann. N. Y. Acad. Sci. 51:660–672.

    Google Scholar 

  14. Blumberg, P. M. 1981. In vitro studies on the mode of action of the phorbol esters, potent tumor promoters, part 2. Crit. Rev. Toxicol. 8:199–234.

    PubMed  Google Scholar 

  15. Driedger, P. E., and Blumberg, P. M. 1980. Specific binding of phorbol ester tumor promoters. Proc. Natl. Acad. Sci. U.S.A. 77:567–571.

    PubMed  Google Scholar 

  16. Dunphy, W. G., Delclos K. B., and Blumberg, P. M. 1980. Characterization of specific binding of [3H]-phorbol 12, 13-dibutyrate and [3H]-phorbol 12-myristate 13-acetate to mouse brain. Cancer Res. 40:3635–3641.

    PubMed  Google Scholar 

  17. Leach, K. L., James, M. L., and Blumberg, P. M. 1983. Characterization of a specific phorbol ester aporeceptor in mouse brain cytosol. Proc. Natl. Acad. Sci. U.S.A. 80:4208–4212.

    PubMed  Google Scholar 

  18. Heiman, A. S., and Crews, F. T. 1985. Characterization of the effects of phorbol esters on rat mast cell secretion. J. Immunol. 134:548–555.

    PubMed  Google Scholar 

  19. Feldstein, J. B., Gonzales, R. A., Baker, S. P., Sumners, C., Crews, F. T., and Raizada, M. K. 1986. Decreased αladrenergic receptor-mediated inositide hydrolysis in neurons from hypertensive rat brain. Am. J. Physiol. 2 251:C230–237.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Raizada, M.K., Morse, C.A., Gonzales, R.A. et al. Receptors for phorbol esters are primarily localized in neurons: Comparison of neuronal and glial cultures. Neurochem Res 13, 51–56 (1988). https://doi.org/10.1007/BF00971854

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00971854

Key Words

Navigation