Abstract
LiCl (2.5-20 mM) reversibly suppressed nerve growth factor (NGF)- induced neurite outgrowth by cultured rat PC 12 pheochromocytoma cells. Similar concentrations of LiCl also reversibly blocked NGF-dependent regeneration of neurites by PC12 cells that had been primed by long- term pre-exposure to NGF and by cultured newborn mouse sympathetic neurons. In contrast, transcription-dependent responses of PC12 cells to NGF such as priming and induction of the NGF-inducible large external glycoprotein, occurred despite the presence of Li+. SDS PAGE analysis of total cellular phosphoproteins (labeled by 2-h exposure to 32P-orthophosphate) from neurite-bearing primed PC12 cells revealed that Li+ reversibly inhibited the phosphorylation of a band of Mr 64,000 that was barely detectable in NGF-untreated PC12 cells. However, Li+ did not appear to affect the labeling of other phosphoproteins in either NGF-primed or untreated PC12 cultures, nor did it affect the rapid increase in phosphorylation of several proteins that occurs when NGF is first added to unprimed cultures. Several criteria indicated that the NGF-inducible phosphoprotein of Mr 64,000 is a microtubule- associated protein (MAP). Of the NGF-inducible phosphorylated MAPs that have been detected in PC12 cells (Mr 64,000, 72,000, 80,000, and 320,000), several (Mr 64,000, 72,000, and 80,000) were found to be substantially less phosphorylated in the presence of Li+. Neither a phorbol ester tumor promotor nor permeant cAMP analogs reversed the inhibitory effects of Li+ on neurite outgrowth or on phosphorylation of the component of Mr 64,000. Microtubules are a major and required constituent of neurites, and MAPs may regulate the assembly and stability of neuritic microtubules. The observation that Li+ selectively inhibits NGF-induced neurite outgrowth and MAP phosphorylation suggests a possible causal relationship between these two events.
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- Allison J. H., Blisner M. E., Holland W. H., Hipps P. P., Sherman W. R. Increased brain myo-inositol 1-phosphate in lithium-treated rats. Biochem Biophys Res Commun. 1976 Jul 26;71(2):664–670. doi: 10.1016/0006-291x(76)90839-1. [DOI] [PubMed] [Google Scholar]
- Apte S. N., Langston J. W. Permanent neurological deficits due to lithium toxicity. Ann Neurol. 1983 Apr;13(4):453–455. doi: 10.1002/ana.410130414. [DOI] [PubMed] [Google Scholar]
- Bernd P., Greene L. A. Association of 125I-nerve growth factor with PC12 pheochromocytoma cells. Evidence for internalization via high-affinity receptors only and for long-term regulation by nerve growth factor of both high- and low-affinity receptors. J Biol Chem. 1984 Dec 25;259(24):15509–15516. [PubMed] [Google Scholar]
- Berridge M. J., Downes C. P., Hanley M. R. Lithium amplifies agonist-dependent phosphatidylinositol responses in brain and salivary glands. Biochem J. 1982 Sep 15;206(3):587–595. doi: 10.1042/bj2060587. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Black M. M., Kurdyla J. T. Microtubule-associated proteins of neurons. J Cell Biol. 1983 Oct;97(4):1020–1028. doi: 10.1083/jcb.97.4.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burstein D. E., Greene L. A. Evidence for RNA synthesis-dependent and -independent pathways in stimulation of neurite outgrowth by nerve growth factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6059–6063. doi: 10.1073/pnas.75.12.6059. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
- Connolly J. L., Greene L. A., Viscarello R. R., Riley W. D. Rapid, sequential changes in surface morphology of PC12 pheochromocytoma cells in response to nerve growth factor. J Cell Biol. 1979 Sep;82(3):820–827. doi: 10.1083/jcb.82.3.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Gard D. L., Kirschner M. W. A polymer-dependent increase in phosphorylation of beta-tubulin accompanies differentiation of a mouse neuroblastoma cell line. J Cell Biol. 1985 Mar;100(3):764–774. doi: 10.1083/jcb.100.3.764. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garrels J. I., Schubert D. Modulation of protein synthesis by nerve growth factor. J Biol Chem. 1979 Aug 25;254(16):7978–7985. [PubMed] [Google Scholar]
- Gaskin F., Cantor C. R., Shelanski M. L. Turbidimetric studies of the in vitro assembly and disassembly of porcine neurotubules. J Mol Biol. 1974 Nov 15;89(4):737–755. doi: 10.1016/0022-2836(74)90048-5. [DOI] [PubMed] [Google Scholar]
- Gelfand E. W., Dosch H. M., Hastings B., Shore A. Lithium: a modulator of cyclic AMP-dependent events in lymphocytes? Science. 1979 Jan 26;203(4378):365–367. doi: 10.1126/science.216075. [DOI] [PubMed] [Google Scholar]
- Gibbons B. H., Gibbons I. R. Lithium reversibly inhibits microtubule-based motility in sperm flagella. Nature. 1984 Jun 7;309(5968):560–562. doi: 10.1038/309560a0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Greene L. A., Shooter E. M. The nerve growth factor: biochemistry, synthesis, and mechanism of action. Annu Rev Neurosci. 1980;3:353–402. doi: 10.1146/annurev.ne.03.030180.002033. [DOI] [PubMed] [Google Scholar]
- 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]
- Halegoua S., Patrick J. Nerve growth factor mediates phosphorylation of specific proteins. Cell. 1980 Nov;22(2 Pt 2):571–581. doi: 10.1016/0092-8674(80)90367-0. [DOI] [PubMed] [Google Scholar]
- Hallcher L. M., Sherman W. R. The effects of lithium ion and other agents on the activity of myo-inositol-1-phosphatase from bovine brain. J Biol Chem. 1980 Nov 25;255(22):10896–10901. [PubMed] [Google Scholar]
- Lee V. M., Shelanski M. L., Greene L. A. Characterization of antisera raised against cultured rat sympathetic neurons. Neuroscience. 1980;5(12):2239–2245. doi: 10.1016/0306-4522(80)90140-2. [DOI] [PubMed] [Google Scholar]
- Levi-Montalcini R., Angeletti P. U. Nerve growth factor. Physiol Rev. 1968 Jul;48(3):534–569. doi: 10.1152/physrev.1968.48.3.534. [DOI] [PubMed] [Google Scholar]
- Lindwall G., Cole R. D. Phosphorylation affects the ability of tau protein to promote microtubule assembly. J Biol Chem. 1984 Apr 25;259(8):5301–5305. [PubMed] [Google Scholar]
- 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]
- McGuire J. C., Greene L. A. Rapid stimulation by nerve growth factor of amino acid uptake by clonal PC12 pheochromocytoma cells. J Biol Chem. 1979 May 10;254(9):3362–3367. [PubMed] [Google Scholar]
- McKeithan T. W., Rosenbaum J. L. The biochemistry of microtubules. A review. Cell Muscle Motil. 1984;5:255–288. doi: 10.1007/978-1-4684-4592-3_7. [DOI] [PubMed] [Google Scholar]
- Mobley W. C., Schenker A., Shooter E. M. Characterization and isolation of proteolytically modified nerve growth factor. Biochemistry. 1976 Dec 14;15(25):5543–5552. doi: 10.1021/bi00670a019. [DOI] [PubMed] [Google Scholar]
- Pallas D., Solomon F. Cytoplasmic microtubule-associated proteins: phosphorylation at novel sites is correlated with their incorporation into assembled microtubules. Cell. 1982 Sep;30(2):407–414. doi: 10.1016/0092-8674(82)90238-0. [DOI] [PubMed] [Google Scholar]
- Pamphlett R. S., Mackenzie R. A. Severe peripheral neuropathy due to lithium intoxication. J Neurol Neurosurg Psychiatry. 1982 Jul;45(7):656–656. doi: 10.1136/jnnp.45.7.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Partlow L. M., Larrabee M. G. Effects of a nerve-growth factor, embryo age and metabolic inhibitors on growth of fibres and on synthesis of ribonucleic acid and protein in embryonic sympathetic ganglia. J Neurochem. 1971 Nov;18(11):2101–2118. doi: 10.1111/j.1471-4159.1971.tb05069.x. [DOI] [PubMed] [Google Scholar]
- Rosenthal N. E., Goodwin F. K. The role of the lithium ion in medicine. Annu Rev Med. 1982;33:555–568. doi: 10.1146/annurev.me.33.020182.003011. [DOI] [PubMed] [Google Scholar]
- Rybak S. M., Stockdale F. E. Growth effects of lithium chloride in BALB/c 3T3 fibroblasts and Madin-Darby canine kidney epithelial cells. Exp Cell Res. 1981 Dec;136(2):263–270. doi: 10.1016/0014-4827(81)90004-5. [DOI] [PubMed] [Google Scholar]
- Seeley P. J., Greene L. A. Short-latency local actions of nerve growth factor at the growth cone. Proc Natl Acad Sci U S A. 1983 May;80(9):2789–2793. doi: 10.1073/pnas.80.9.2789. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sherman W. R., Leavitt A. L., Honchar M. P., Hallcher L. M., Phillips B. E. Evidence that lithium alters phosphoinositide metabolism: chronic administration elevates primarily D-myo-inositol-1-phosphate in cerebral cortex of the rat. J Neurochem. 1981 Jun;36(6):1947–1951. doi: 10.1111/j.1471-4159.1981.tb10819.x. [DOI] [PubMed] [Google Scholar]
- Solomon F., Magendantz M., Salzman A. Identification with cellular microtubules of one of the co-assemlbing microtubule-associated proteins. Cell. 1979 Oct;18(2):431–438. doi: 10.1016/0092-8674(79)90062-x. [DOI] [PubMed] [Google Scholar]
- Sternlicht H., Ringel I. Colchicine inhibition of microtubule assembly via copolymer formation. J Biol Chem. 1979 Oct 25;254(20):10540–10550. [PubMed] [Google Scholar]
- Tomooka Y., Imagawa W., Nandi S., Bern H. A. Growth effect of lithium on mouse mammary epithelial cells in serum-free collagen gel culture. J Cell Physiol. 1983 Dec;117(3):290–296. doi: 10.1002/jcp.1041170303. [DOI] [PubMed] [Google Scholar]
- Uchigata M., Tanabe H., Hasue I., Kurihara M. Peripheral neuropathy due to lithium intoxication. Ann Neurol. 1981 Apr;9(4):414–414. doi: 10.1002/ana.410090421. [DOI] [PubMed] [Google Scholar]
- Verma D. S., Spitzer G., Gutterman J. U., Beran M., Zander A. R., McCredie K. B. Human leukocyte interferon-mediated granulopoietic differentiation arrest and its abrogation by lithium carbonate. Am J Hematol. 1982 Feb;12(1):39–46. doi: 10.1002/ajh.2830120106. [DOI] [PubMed] [Google Scholar]
- Wolff J., Berens S. C., Jones A. B. Inhibition of thyrotropin-stimulated adenyl cyclase activity of beef thyroid membranes by low concentration of lithium ion. Biochem Biophys Res Commun. 1970 Apr 8;39(1):77–82. doi: 10.1016/0006-291x(70)90760-6. [DOI] [PubMed] [Google Scholar]
- Yu M. W., Tolson N. W., Guroff G. Increased phosphorylation of specific nuclear proteins in superior cervical ganglia and PC12 cells in response to nerve growth factor. J Biol Chem. 1980 Nov 10;255(21):10481–10492. [PubMed] [Google Scholar]
- Zieve G., Solomon F. Direct isolation of neuronal microtubule skeletons. Mol Cell Biol. 1984 Feb;4(2):371–374. doi: 10.1128/mcb.4.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]