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. 1988 Aug 1;107(2):643–650. doi: 10.1083/jcb.107.2.643

PC12 cells express juvenile microtubule-associated proteins during nerve growth factor-induced neurite outgrowth

PMCID: PMC2115230  PMID: 3417766

Abstract

Microtubule-associated proteins (MAPs) are believed to play an important role in regulating the growth of neuronal processes. The nerve growth factor-induced differentiation of PC12 pheochromocytoma cells is a widely used tissue culture model for studying this mechanism. We have found that contrary to previous suggestions, the major MAPs of adult brain, MAP1 and MAP2, are minor components of PC12 cells. Instead two novel MAPs characteristic of developing brain, MAP3 and MAP5, are present and increase more than 10-fold after nerve growth factor treatment; the timing of these increases coinciding with the bundling of microtubules and neurite outgrowth. Immunocytochemical staining showed that MAP3 and MAP5 are initially distributed throughout the cytoplasm. Subsequently MAP5 becomes associated with microtubules in both neurites and growth cones but MAP3 distribution remained diffuse. Thus MAP3 and MAP5, which are characteristic of developing neurons in the juvenile brain, are also induced in PC12 cells during neurite outgrowth in culture. In contrast MAP1, which is characteristic of mature neurons, does not increase during PC12 cell differentiation. These results provide evidence that one set of MAPs is expressed during neurite outgrowth and a different set during the maintenance of neuronal form. It also appears that the PC12 system is an appropriate model for studying the active neurite growth phase of neuronal differentiation but not for neuronal maturation.

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Selected References

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  1. Bernhardt R., Huber G., Matus A. Differences in the developmental patterns of three microtubule-associated proteins in the rat cerebellum. J Neurosci. 1985 Apr;5(4):977–991. doi: 10.1523/JNEUROSCI.05-04-00977.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Binder L. I., Frankfurter A., Rebhun L. I. The distribution of tau in the mammalian central nervous system. J Cell Biol. 1985 Oct;101(4):1371–1378. doi: 10.1083/jcb.101.4.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Black M. M., Aletta J. M., Greene L. A. Regulation of microtubule composition and stability during nerve growth factor-promoted neurite outgrowth. J Cell Biol. 1986 Aug;103(2):545–557. doi: 10.1083/jcb.103.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Cleveland D. W., Hwo S. Y., Kirschner M. W. Physical and chemical properties of purified tau factor and the role of tau in microtubule assembly. J Mol Biol. 1977 Oct 25;116(2):227–247. doi: 10.1016/0022-2836(77)90214-5. [DOI] [PubMed] [Google Scholar]
  6. Daniels M. P. Colchicine inhibition of nerve fiber formation in vitro. J Cell Biol. 1972 Apr;53(1):164–176. doi: 10.1083/jcb.53.1.164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drubin D. G., Feinstein S. C., Shooter E. M., Kirschner M. W. Nerve growth factor-induced neurite outgrowth in PC12 cells involves the coordinate induction of microtubule assembly and assembly-promoting factors. J Cell Biol. 1985 Nov;101(5 Pt 1):1799–1807. doi: 10.1083/jcb.101.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fellous A., Francon J., Lennon A. M., Nunez J. Microtubule assembly in vitro. Purification of assembly-promoting factors. Eur J Biochem. 1977 Aug 15;78(1):167–174. doi: 10.1111/j.1432-1033.1977.tb11726.x. [DOI] [PubMed] [Google Scholar]
  9. Francon J., Lennon A. M., Fellous A., Mareck A., Pierre M., Nunez J. Heterogeneity of microtubule-associated proteins and brain development. Eur J Biochem. 1982 Dec 15;129(2):465–471. doi: 10.1111/j.1432-1033.1982.tb07072.x. [DOI] [PubMed] [Google Scholar]
  10. Garner C. C., Brugg B., Matus A. A 70-kilodalton microtubule-associated protein (MAP2c), related to MAP2. J Neurochem. 1988 Feb;50(2):609–615. doi: 10.1111/j.1471-4159.1988.tb02954.x. [DOI] [PubMed] [Google Scholar]
  11. Garner C. C., Matus A. Different forms of microtubule-associated protein 2 are encoded by separate mRNA transcripts. J Cell Biol. 1988 Mar;106(3):779–783. doi: 10.1083/jcb.106.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greene L. A., Burstein D. E., Black M. M. The role of transcription-dependent priming in nerve growth factor promoted neurite outgrowth. Dev Biol. 1982 Jun;91(2):305–316. doi: 10.1016/0012-1606(82)90037-9. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  16. Heidemann S. R., Joshi H. C., Schechter A., Fletcher J. R., Bothwell M. Synergistic effects of cyclic AMP and nerve growth factor on neurite outgrowth and microtubule stability of PC12 cells. J Cell Biol. 1985 Mar;100(3):916–927. doi: 10.1083/jcb.100.3.916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huber G., Alaimo-Beuret D., Matus A. MAP3: characterization of a novel microtubule-associated protein. J Cell Biol. 1985 Feb;100(2):496–507. doi: 10.1083/jcb.100.2.496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Huber G., Matus A. Differences in the cellular distributions of two microtubule-associated proteins, MAP1 and MAP2, in rat brain. J Neurosci. 1984 Jan;4(1):151–160. doi: 10.1523/JNEUROSCI.04-01-00151.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Huber G., Pehling G., Matus A. The novel microtubule-associated protein MAP3 contributes to the in vitro assembly of brain microtubules. J Biol Chem. 1986 Feb 15;261(5):2270–2273. [PubMed] [Google Scholar]
  20. Jacobs J. R., Stevens J. K. Changes in the organization of the neuritic cytoskeleton during nerve growth factor-activated differentiation of PC12 cells: a serial electron microscopic study of the development and control of neurite shape. J Cell Biol. 1986 Sep;103(3):895–906. doi: 10.1083/jcb.103.3.895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jacobs J. R., Stevens J. K. Experimental modification of PC12 neurite shape with the microtubule-depolymerizing drug Nocodazole: a serial electron microscopic study of neurite shape control. J Cell Biol. 1986 Sep;103(3):907–915. doi: 10.1083/jcb.103.3.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Joshi H. C., Chu D., Buxbaum R. E., Heidemann S. R. Tension and compression in the cytoskeleton of PC 12 neurites. J Cell Biol. 1985 Sep;101(3):697–705. doi: 10.1083/jcb.101.3.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Keates R. A., Hall R. H. Tubulin requires an accessory protein for self assembly in microtubules. Nature. 1975 Oct 2;257(5525):418–421. doi: 10.1038/257418a0. [DOI] [PubMed] [Google Scholar]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. Luckenbill-Edds L., Van Horn C., Greene L. A. Fine structure of initial outgrowth of processes induced in a pheochromocytoma cell line (PC12) by nerve growth factor. J Neurocytol. 1979 Aug;8(4):493–511. doi: 10.1007/BF01214805. [DOI] [PubMed] [Google Scholar]
  26. Magendantz M., Solomon F. Analyzing the components of microtubules: antibodies against chartins, associated proteins from cultured cells. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6581–6585. doi: 10.1073/pnas.82.19.6581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mareck A., Fellous A., Francon J., Nunez J. Changes in composition and activity of microtubule-associated proteins during brain development. Nature. 1980 Mar 27;284(5754):353–355. doi: 10.1038/284353a0. [DOI] [PubMed] [Google Scholar]
  28. Matus A., Bernhardt R., Hugh-Jones T. High molecular weight microtubule-associated proteins are preferentially associated with dendritic microtubules in brain. Proc Natl Acad Sci U S A. 1981 May;78(5):3010–3014. doi: 10.1073/pnas.78.5.3010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Matus A. Microtubule-associated proteins: their potential role in determining neuronal morphology. Annu Rev Neurosci. 1988;11:29–44. doi: 10.1146/annurev.ne.11.030188.000333. [DOI] [PubMed] [Google Scholar]
  30. Matus A., Riederer B. Microtubule-associated proteins in the developing brain. Ann N Y Acad Sci. 1986;466:167–179. doi: 10.1111/j.1749-6632.1986.tb38393.x. [DOI] [PubMed] [Google Scholar]
  31. Murphy D. B., Borisy G. G. Association of high-molecular-weight proteins with microtubules and their role in microtubule assembly in vitro. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2696–2700. doi: 10.1073/pnas.72.7.2696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nunez J. Differential expression of microtubule components during brain development. Dev Neurosci. 1986;8(3):125–141. doi: 10.1159/000112248. [DOI] [PubMed] [Google Scholar]
  33. Riederer B., Cohen R., Matus A. MAP5: a novel brain microtubule-associated protein under strong developmental regulation. J Neurocytol. 1986 Dec;15(6):763–775. doi: 10.1007/BF01625193. [DOI] [PubMed] [Google Scholar]
  34. Riederer B., Matus A. Differential expression of distinct microtubule-associated proteins during brain development. Proc Natl Acad Sci U S A. 1985 Sep;82(17):6006–6009. doi: 10.1073/pnas.82.17.6006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Seeds N. W., Gilman A. G., Amano T., Nirenberg M. W. Regulation of axon formation by clonal lines of a neural tumor. Proc Natl Acad Sci U S A. 1970 May;66(1):160–167. doi: 10.1073/pnas.66.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tucker R. P., Binder L. I., Matus A. I. Neuronal microtubule-associated proteins in the embryonic avian spinal cord. J Comp Neurol. 1988 May 1;271(1):44–55. doi: 10.1002/cne.902710106. [DOI] [PubMed] [Google Scholar]
  37. Tucker R. P., Matus A. I. Developmental regulation of two microtubule-associated proteins (MAP2 and MAP5) in the embryonic avian retina. Development. 1987 Nov;101(3):535–546. doi: 10.1242/dev.101.3.535. [DOI] [PubMed] [Google Scholar]
  38. Vallee R. B. A taxol-dependent procedure for the isolation of microtubules and microtubule-associated proteins (MAPs). J Cell Biol. 1982 Feb;92(2):435–442. doi: 10.1083/jcb.92.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weingarten M. D., Lockwood A. H., Hwo S. Y., Kirschner M. W. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. 1975 May;72(5):1858–1862. doi: 10.1073/pnas.72.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yamada K. M., Spooner B. S., Wessells N. K. Axon growth: roles of microfilaments and microtubules. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1206–1212. doi: 10.1073/pnas.66.4.1206. [DOI] [PMC free article] [PubMed] [Google Scholar]

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