• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. May 1988; 85(10): 3440–3444.
PMCID: PMC280227

Thrombin modulates and reverses neuroblastoma neurite outgrowth.


Previous studies have shown that neuroblastoma cells and several types of primary neuronal cells in culture rapidly extend neurites when switched from serum-containing to serum-free medium. The present studies on cloned neuroblastoma cells show that thrombin blocked this spontaneous differentiation at 2 nM with a half-maximal potency of 50 pM. This required the catalytic activity of thrombin and was reversed upon thrombin removal. Thrombin also caused cells in serum-free medium to retract their neurites at equally low concentrations. Two other serine proteases, urokinase and plasmin, did not block or reverse neurite extension even at 100-fold higher concentrations. A specific assay for thrombin indicated that thrombin detected in serum-containing medium from neuroblastoma cultures was derived from serum and that it was likely responsible for much of the known capacity of serum to maintain neuroblastoma cells in a nondifferentiated state. This was supported by the finding that heparin addition reduced the thrombin concentration in serum-containing medium and stimulated neurite outgrowth from neuroblastoma cells in serum-containing medium. Studies on the ability of thrombin to modulate neurite outgrowth by other agents showed that it blocked and reversed the neurite outgrowth activity of two thrombin inhibitors: protease nexin-1 (which is identical to glial-derived neurite-promoting factor) and hirudin. Thrombin, however, did not block the neurite-promoting activity of dibutyryl cAMP or prostaglandin E1. These results suggest a specific role for thrombin in control of neurite outgrowth.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Furmanski P, Silverman DJ, Lubin M. Expression of differentiated functions in mouse neuroblastoma mediated by dibutyryl-cyclic adenosine monophosphate. Nature. 1971 Oct 8;233(5319):413–415. [PubMed]
  • Prasad KN, Hsie AW. Morphologic differentiation of mouse neuroblastoma cells induced in vitro by dibutyryl adenosine 3':5'-cyclic monophosphate. Nat New Biol. 1971 Sep 29;233(39):141–142. [PubMed]
  • Prasad KN, Sahu SK, Sinha PK. Cyclic nucleotides in the regulation of expression of differentiated functions in neuroblastoma cells. J Natl Cancer Inst. 1976 Sep;57(3):619–631. [PubMed]
  • Prasad KN. Morphological differentiation induced by prostaglandin in mouse neuroblastoma cells in culture. Nat New Biol. 1972 Mar 15;236(63):49–52. [PubMed]
  • Bottenstein JE, Sato GH. Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A. 1979 Jan;76(1):514–517. [PMC free article] [PubMed]
  • Schubert D, Humphreys S, Baroni C, Cohn M. In vitro differentiation of a mouse neuroblastoma. Proc Natl Acad Sci U S A. 1969 Sep;64(1):316–323. [PMC free article] [PubMed]
  • Schubert D, Humphreys S, Jacob F, de Vitry F. Induced differentiation of a neuroblastoma. Dev Biol. 1971 Aug;25(4):514–546. [PubMed]
  • Schubert D, Heinemann S, Carlisle W, Tarikas H, Kimes B, Patrick J, Steinbach JH, Culp W, Brandt BL. Clonal cell lines from the rat central nervous system. Nature. 1974 May 17;249(454):224–227. [PubMed]
  • Seeds NW, Gilman AG, Amano T, Nirenberg MW. Regulation of axon formation by clonal lines of a neural tumor. Proc Natl Acad Sci U S A. 1970 May;66(1):160–167. [PMC free article] [PubMed]
  • Marchisio PC, Osborn M, Weber K. The intracellular organization of actin and tubulin in cultured C-1300 mouse neuroblastoma cells (clone NB41A3). J Neurocytol. 1978 Oct;7(5):571–582. [PubMed]
  • Olmsted JB. Tubulin pools in differentiating neuroblastoma cells. J Cell Biol. 1981 Jun;89(3):418–423. [PMC free article] [PubMed]
  • Amano T, Richelson E, Nirenberg M. Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase-acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc Natl Acad Sci U S A. 1972 Jan;69(1):258–263. [PMC free article] [PubMed]
  • Augusti-Tocco G, Sato G. Establishment of functional clonal lines of neurons from mouse neuroblastoma. Proc Natl Acad Sci U S A. 1969 Sep;64(1):311–315. [PMC free article] [PubMed]
  • Schurch-Rathgeb Y, Mongard D. Brain development influences the appearance of glial factor-like activity in rat brain primary cultures. Nature. 1978 May 25;273(5660):308–309. [PubMed]
  • Monard D, Niday E, Limat A, Solomon F. Inhibition of protease activity can lead to neurite extension in neuroblastoma cells. Prog Brain Res. 1983;58:359–364. [PubMed]
  • Guenther J, Nick H, Monard D. A glia-derived neurite-promoting factor with protease inhibitory activity. EMBO J. 1985 Aug;4(8):1963–1966. [PMC free article] [PubMed]
  • Stone SR, Nick H, Hofsteenge J, Monard D. Glial-derived neurite-promoting factor is a slow-binding inhibitor of trypsin, thrombin, and urokinase. Arch Biochem Biophys. 1987 Jan;252(1):237–244. [PubMed]
  • Krystosek A, Seeds NW. Plasminogen activator release at the neuronal growth cone. Science. 1981 Sep 25;213(4515):1532–1534. [PubMed]
  • Krystosek A, Seeds NW. Plasminogen activator secretion by granule neurons in cultures of developing cerebellum. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7810–7814. [PMC free article] [PubMed]
  • Soreq H, Miskin R, Zutra A, Littauer UZ. Modulation in the levels and localization of plasminogen activator in differentiating neuroblastoma cells. Brain Res. 1983 Apr;283(2-3):257–269. [PubMed]
  • Snider RM, McKinney M, Fenton JW, 2nd, Richelson E. Activation of cyclic nucleotide formation in murine neuroblastoma N1E-115 cells by modified human thrombins. J Biol Chem. 1984 Jul 25;259(14):9078–9081. [PubMed]
  • Snider RM, McKinney M, Richelson E. Thrombin binding and stimulation of cyclic guanosine monophosphate formation in neuroblastoma cells. Semin Thromb Hemost. 1986 Jul;12(3):253–262. [PubMed]
  • Snider RM. Thrombin effects on cultured nerve cells: clinical implications and evidence for a novel mechanism of neuronal activation. Ann N Y Acad Sci. 1986;485:310–313. [PubMed]
  • Low DA, Cunningham DD. A novel method for measuring cell surface-bound thrombin. Detection of iodination-induced changes in thrombin-binding affinity. J Biol Chem. 1982 Jan 25;257(2):850–858. [PubMed]
  • Farrell DH, Van Nostrand WE, Cunningham DD. A simple two-step purification of protease nexin. Biochem J. 1986 Aug 1;237(3):907–912. [PMC free article] [PubMed]
  • Adler R. Regulation of neurite growth in purified retina neuronal cultures: effects of PNPF, a substratum-bound, neurite-promoting factor. J Neurosci Res. 1982;8(2-3):165–177. [PubMed]
  • Davis GE, Skaper SD, Manthorpe M, Moonen G, Varon S. Fetal calf serum-mediated inhibition of neurite growth from ciliary ganglion neurons in vitro. J Neurosci Res. 1984;12(1):29–39. [PubMed]
  • Ludueña MA. The growth of spinal ganglion neurons in serum-free medium. Dev Biol. 1973 Aug;33(2):470–476. [PubMed]
  • Ziller C, Le Douarin NM. Neuronal differentiation in cultured neural crest cells: the effect of serum on neurite outgrowth. Birth Defects Orig Artic Ser. 1983;19(4):251–261. [PubMed]
  • Ziller C, Dupin E, Brazeau P, Paulin D, Le Douarin NM. Early segregation of a neuronal precursor cell line in the neural crest as revealed by culture in a chemically defined medium. Cell. 1983 Feb;32(2):627–638. [PubMed]
  • Gloor S, Odink K, Guenther J, Nick H, Monard D. A glia-derived neurite promoting factor with protease inhibitory activity belongs to the protease nexins. Cell. 1986 Dec 5;47(5):687–693. [PubMed]
  • Rosenblatt DE, Cotman CW, Nieto-Sampedro M, Rowe JW, Knauer DJ. Identification of a protease inhibitor produced by astrocytes that is structurally and functionally homologous to human protease nexin-I. Brain Res. 1987 Jul 7;415(1):40–48. [PubMed]
  • Baker JB, Low DA, Simmer RL, Cunningham DD. Protease-nexin: a cellular component that links thrombin and plasminogen activator and mediates their binding to cells. Cell. 1980 Aug;21(1):37–45. [PubMed]
  • Means ED, Anderson DK. Thrombin interactions with central nervous system tissue and implications of these interactions. Ann N Y Acad Sci. 1986;485:314–322. [PubMed]
  • McKinney M, Snider RM, Richelson E. Thrombin binding to human brain and spinal cord. Mayo Clin Proc. 1983 Dec;58(12):829–831. [PubMed]

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


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...