Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Aug;82(15):5223–5226. doi: 10.1073/pnas.82.15.5223

Peripheral-type benzodiazepines influence ornithine decarboxylase levels and neurite outgrowth in PC12 cells.

J I Morgan, M D Johnson, J K Wang, K H Sonnenfeld, S Spector
PMCID: PMC390532  PMID: 2991912

Abstract

A number of the benzodiazepines (BZDs) inhibit nerve growth factor (NGF)-induced neurite outgrowth in a dose-dependent manner in PC12 cell cultures. The rank order of potency of a series of BZDs for inhibition of neurite outgrowth does not correlate with the order of their affinity constants for the so-called peripheral BZD sites present on PC12 cells. Whereas the inhibition of neurite extension is stereospecific, the binding to the peripheral site is not. The inhibition of neurite outgrowth is not attributable to a blockade of NGF binding to either its fast or slow receptors. Additionally, several other characteristics of NGF stimulation of PC12 cells, such as autoadhesion and the induction of ornithine decarboxylase (OrnDCase), remain unaltered in the presence of BZDs. Many BZDs increase OrnDCase levels in PC12 cells in the absence of NGF. The OrnDCase response to BZDs is blocked by actinomycin D. Furthermore, the structural requirements of BZDs for induction of OrnDCase activity is not identical to that for the inhibition of NGF-induced neurite extension. Thus, unlike attenuation of neurite extension, BZD induction of OrnDCase is not stereospecific. Also, some BZDs block neurite growth but do not induce OrnDCase. We propose that there are at least three sites of action of BZDs on PC12 cells. The first is the well-characterized high-affinity peripheral site. The second distinct locus of action results in a blockade of NGF-induced neurite outgrowth. The structural requirements for the latter effect are essentially indistinguishable from those for BZD induction of hemoglobin synthesis in Friend erythroleukemia cells. The third site of action results in an induction of OrnDCase. Structural requirements for this activity are not identical to those for either the differentiation of Friend cells or the inhibition of neurite outgrowth.

Full text

PDF
5223

Images in this article

5225Image
on p.5225
5225Image
on p.5225
5225Image
on p.5225
5225Image
on p.5225

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bandman E., Walker C. R., Strohman R. C. Diazepam inhibits myoblast fusion and expression of muscle specific protein synthesis. Science. 1978 May 5;200(4341):559–561. doi: 10.1126/science.565534. [DOI] [PubMed] [Google Scholar]
  2. Black M. M., Greene L. A. Changes in the colchicine susceptibility of microtubules associated with neurite outgrowth: studies with nerve growth factor-responsive PC12 pheochromocytoma cells. J Cell Biol. 1982 Nov;95(2 Pt 1):379–386. doi: 10.1083/jcb.95.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowling A. C., DeLorenzo R. J. Micromolar affinity benzodiazepine receptors: identification and characterization in central nervous system. Science. 1982 Jun 11;216(4551):1247–1250. doi: 10.1126/science.6281893. [DOI] [PubMed] [Google Scholar]
  4. Braestrup C., Squires R. F. Specific benzodiazepine receptors in rat brain characterized by high-affinity (3H)diazepam binding. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3805–3809. doi: 10.1073/pnas.74.9.3805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burton L. E., Wilson W. H., Shooter E. M. Nerve growth factor in mouse saliva. Rapid isolation procedures for and characterization of 7 S nerve growth factor. J Biol Chem. 1978 Nov 10;253(21):7807–7812. [PubMed] [Google Scholar]
  6. Clarke G. D., Ryan P. J. Tranquillizers can block mitogenesis in 3T3 cells and induce differentiation in Friend cells. Nature. 1980 Sep 11;287(5778):160–161. doi: 10.1038/287160a0. [DOI] [PubMed] [Google Scholar]
  7. Friend C., Scher W., Holland J. G., Sato T. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide. Proc Natl Acad Sci U S A. 1971 Feb;68(2):378–382. doi: 10.1073/pnas.68.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Greene L. A., Liem R. K., Shelanski M. L. Regulation of a high molecular weight microtubule-associated protein in PC12 cells by nerve growth factor. J Cell Biol. 1983 Jan;96(1):76–83. doi: 10.1083/jcb.96.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greene L. A., McGuire J. C. Induction of ornithine decarboxylase by nerve growth factor dissociated from effects on survival and neurite outgrowth. Nature. 1978 Nov 9;276(5684):191–194. doi: 10.1038/276191a0. [DOI] [PubMed] [Google Scholar]
  10. Greene L. A., Tischler A. S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424–2428. doi: 10.1073/pnas.73.7.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guroff G., Dickens G., End D. The induction of ornithine decarboxylase by nerve growth factor and epidermal growth factor in PC12 cells. J Neurochem. 1981 Aug;37(2):342–349. doi: 10.1111/j.1471-4159.1981.tb00461.x. [DOI] [PubMed] [Google Scholar]
  12. Guroff G., Dickens G. The effects of nerve growth factor on polyamine metabolism in PC12 cells. J Neurochem. 1983 May;40(5):1271–1277. doi: 10.1111/j.1471-4159.1983.tb13566.x. [DOI] [PubMed] [Google Scholar]
  13. Gusella J. F., Housman D. Induction of erythroid differentiation in vitro by purines and purine analogues. Cell. 1976 Jun;8(2):263–269. doi: 10.1016/0092-8674(76)90010-6. [DOI] [PubMed] [Google Scholar]
  14. Hatanaka H., Otten U., Thoenen H. Nerve growth factor-mediated selective induction of ornithine decarboxylase in rat pheochromocytoma; a cyclic AMP-independent process. FEBS Lett. 1978 Aug 15;92(2):313–316. doi: 10.1016/0014-5793(78)80777-7. [DOI] [PubMed] [Google Scholar]
  15. Huff K. R., Guroff G. Nerve growth factor-induced reduction in epidermal growth factor responsiveness and epidermal growth factor receptors in PC12 cells: an aspect of cell differentiation. Biochem Biophys Res Commun. 1979 Jul 12;89(1):175–180. doi: 10.1016/0006-291x(79)90960-4. [DOI] [PubMed] [Google Scholar]
  16. Marangos P. J., Patel J., Boulenger J. P., Clark-Rosenberg R. Characterization of peripheral-type benzodiazepine binding sites in brain using [3H]Ro 5-4864. Mol Pharmacol. 1982 Jul;22(1):26–32. [PubMed] [Google Scholar]
  17. Matthew E., Laskin J. D., Zimmerman E. A., Weinstein I. B., Hsu K. C., Engelhardt D. L. Benzodiazepines have high-affinity binding sites and induce melanogenesis in B16/C3 melanoma cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3935–3939. doi: 10.1073/pnas.78.6.3935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Möhler H., Okada T. Benzodiazepine receptor: demonstration in the central nervous system. Science. 1977 Nov 25;198(4319):849–851. doi: 10.1126/science.918669. [DOI] [PubMed] [Google Scholar]
  19. Pegg A. E., Williams-Ashman H. G. Biosynthesis of putrescine in the prostate gland of the rat. Biochem J. 1968 Jul;108(4):533–539. doi: 10.1042/bj1080533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reuben R. C., Wife R. L., Breslow R., Rifkind R. A., Marks P. A. A new group of potent inducers of differentiation in murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1976 Mar;73(3):862–866. doi: 10.1073/pnas.73.3.862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schechter A. L., Bothwell M. A. Nerve growth factor receptors on PC12 cells: evidence for two receptor classes with differing cytoskeletal association. Cell. 1981 Jun;24(3):867–874. doi: 10.1016/0092-8674(81)90112-4. [DOI] [PubMed] [Google Scholar]
  22. Schoemaker H., Boles R. G., Horst W. D., Yamamura H. I. Specific high-affinity binding sites for [3H]Ro 5-4864 in rat brain and kidney. J Pharmacol Exp Ther. 1983 Apr;225(1):61–69. [PubMed] [Google Scholar]
  23. Seeley P. J., Rukenstein A., Connolly J. L., Greene L. A. Differential inhibition of nerve growth factor and epidermal growth factor effects on the PC12 pheochromocytoma line. J Cell Biol. 1984 Feb;98(2):417–426. doi: 10.1083/jcb.98.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sonnenfeld K. H., Ishii D. N. Nerve growth factor effects and receptors in cultured human neuroblastoma cell lines. J Neurosci Res. 1982;8(2-3):375–391. doi: 10.1002/jnr.490080226. [DOI] [PubMed] [Google Scholar]
  25. Syapin P. J., Skolnick P. Characterization of benzodiazepine binding sites in cultured cells of neural origin. J Neurochem. 1979 Mar;32(3):1047–1051. doi: 10.1111/j.1471-4159.1979.tb04592.x. [DOI] [PubMed] [Google Scholar]
  26. Tallman J. F., Paul S. M., Skolnick P., Gallager D. W. Receptors for the age of anxiety: pharmacology of the benzodiazepines. Science. 1980 Jan 18;207(4428):274–281. doi: 10.1126/science.6101294. [DOI] [PubMed] [Google Scholar]
  27. Taniguchi T., Wang J. K., Spector S. [3H]Diazepam binding sites on rat heart and kidney. Biochem Pharmacol. 1982 Feb 15;31(4):589–590. doi: 10.1016/0006-2952(82)90164-2. [DOI] [PubMed] [Google Scholar]
  28. Varon S., Normura J., Shooter E. M. Reversible dissociation of the mouse nerve growth factor protein into different subunits. Biochemistry. 1968 Apr;7(4):1296–1303. doi: 10.1021/bi00844a008. [DOI] [PubMed] [Google Scholar]
  29. Walker C. R., Bandman E., Strohman R. C. Diazepam induces relaxation of chick embryo muscle fibers in vitro and inhibits myosin synthesis. Exp Cell Res. 1979 Oct 15;123(2):285–291. doi: 10.1016/0014-4827(79)90470-1. [DOI] [PubMed] [Google Scholar]
  30. Wang J. K., Morgan J. I., Spector S. Benzodiazepines that bind at peripheral sites inhibit cell proliferation. Proc Natl Acad Sci U S A. 1984 Feb;81(3):753–756. doi: 10.1073/pnas.81.3.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wang J. K., Morgan J. I., Spector S. Differentiation of Friend erythroleukemia cells induced by benzodiazepines. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3770–3772. doi: 10.1073/pnas.81.12.3770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wang J. K., Taniguchi T., Spector S. Properties of [3H]diazepam binding sites on rat blood platelets. Life Sci. 1980 Nov 17;27(20):1881–1888. doi: 10.1016/0024-3205(80)90434-8. [DOI] [PubMed] [Google Scholar]
  33. Wang J. K., Taniguchi T., Spector S. Structural requirements for the binding of benzodiazepines to their peripheral-type sites. Mol Pharmacol. 1984 May;25(3):349–351. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES