Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. Jul 1, 1992; 118(1): 139–148.
PMCID: PMC2289531

Synthesis and localization of ciliary neurotrophic factor in the sciatic nerve of the adult rat after lesion and during regeneration

Abstract

Ciliary neurotrophic factor (CNTF) is expressed in high quantities in Schwann cells of peripheral nerves during postnatal development of the rat. The absence of a hydrophobic leader sequence and the immunohistochemical localization of CNTF within the cytoplasm of these cells indicate that the factor might not be available to responsive neurons under physiological conditions. However, CNTF supports the survival of a variety of embryonic neurons, including spinal motoneurons in culture. Moreover we have recently demonstrated that the exogenous application of CNTF protein to the lesioned facial nerve of the newborn rat rescued these motoneurons from cell death. These results indicate that CNTF might indeed play a major role in assisting the survival of lesioned neurons in the adult peripheral nervous system. Here we demonstrate that the CNTF mRNA and protein levels and the manner in which they are regulated are compatible with such a function in lesioned peripheral neurons. In particular, immunohistochemical analysis showed significant quantities of CNTF at extracellular sites after sciatic nerve lesion. Western blots and determination of CNTF biological activity of the same nerve segments indicate that extracellular CNTF seems to be biologically active. After nerve lesion CNTF mRNA levels were reduced to less than 5% in distal regions of the sciatic nerve whereas CNTF bioactivity decreased to only one third of the original before-lesion levels. A gradual reincrease in Schwann cells occurred concomitant with regeneration.

Full Text

The Full Text of this article is available as a PDF (2.4M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adler R, Landa KB, Manthorpe M, Varon S. Cholinergic neuronotrophic factors: intraocular distribution of trophic activity for ciliary neurons. Science. 1979 Jun 29;204(4400):1434–1436. [PubMed]
  • Arakawa Y, Sendtner M, Thoenen H. Survival effect of ciliary neurotrophic factor (CNTF) on chick embryonic motoneurons in culture: comparison with other neurotrophic factors and cytokines. J Neurosci. 1990 Nov;10(11):3507–3515. [PubMed]
  • Barbin G, Manthorpe M, Varon S. Purification of the chick eye ciliary neuronotrophic factor. J Neurochem. 1984 Nov;43(5):1468–1478. [PubMed]
  • Barde YA. The nerve growth factor family. Prog Growth Factor Res. 1990;2(4):237–248. [PubMed]
  • Bothwell M. Tissue localization of nerve growth factor and nerve growth factor receptors. Curr Top Microbiol Immunol. 1991;165:55–70. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brown MC, Perry VH, Lunn ER, Gordon S, Heumann R. Macrophage dependence of peripheral sensory nerve regeneration: possible involvement of nerve growth factor. Neuron. 1991 Mar;6(3):359–370. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Duchen LW, Strich SJ. An hereditary motor neurone disease with progressive denervation of muscle in the mouse: the mutant 'wobbler'. J Neurol Neurosurg Psychiatry. 1968 Dec;31(6):535–542. [PMC free article] [PubMed]
  • Eckenstein FP, Esch F, Holbert T, Blacher RW, Nishi R. Purification and characterization of a trophic factor for embryonic peripheral neurons: comparison with fibroblast growth factors. Neuron. 1990 Apr;4(4):623–631. [PubMed]
  • Eckenstein FP, Shipley GD, Nishi R. Acidic and basic fibroblast growth factors in the nervous system: distribution and differential alteration of levels after injury of central versus peripheral nerve. J Neurosci. 1991 Feb;11(2):412–419. [PubMed]
  • Elde R, Cao YH, Cintra A, Brelje TC, Pelto-Huikko M, Junttila T, Fuxe K, Pettersson RF, Hökfelt T. Prominent expression of acidic fibroblast growth factor in motor and sensory neurons. Neuron. 1991 Sep;7(3):349–364. [PubMed]
  • Helfand SL, Smith GA, Wessells NK. Survival and development in culture of dissociated parasympathetic neurons from ciliary ganglia. Dev Biol. 1976 Jun;50(2):541–547. [PubMed]
  • Heumann R, Thoenen H. Comparison between the time course of changes in nerve growth factor protein levels and those of its messenger RNA in the cultured rat iris. J Biol Chem. 1986 Jul 15;261(20):9246–9249. [PubMed]
  • Heumann R, Korsching S, Bandtlow C, Thoenen H. Changes of nerve growth factor synthesis in nonneuronal cells in response to sciatic nerve transection. J Cell Biol. 1987 Jun;104(6):1623–1631. [PMC free article] [PubMed]
  • Heumann R, Lindholm D, Bandtlow C, Meyer M, Radeke MJ, Misko TP, Shooter E, Thoenen H. Differential regulation of mRNA encoding nerve growth factor and its receptor in rat sciatic nerve during development, degeneration, and regeneration: role of macrophages. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8735–8739. [PMC free article] [PubMed]
  • Hughes SM, Lillien LE, Raff MC, Rohrer H, Sendtner M. Ciliary neurotrophic factor induces type-2 astrocyte differentiation in culture. Nature. 1988 Sep 1;335(6185):70–73. [PubMed]
  • Kaupmann Klemens, Sendtner Michael, Stöckli Kurt A, Jockusch Harald. The Gene for Ciliary Neurotrophic Factor (CNTF) Maps to Murine Chromosome 19 and its Expression is Not Affected in the Hereditary Motoneuron Disease 'Wobbler' of the Mouse. Eur J Neurosci. 1991 Oct;3(11):1182–1186. [PubMed]
  • Korsching S, Thoenen H. Quantitative demonstration of the retrograde axonal transport of endogenous nerve growth factor. Neurosci Lett. 1983 Aug 19;39(1):1–4. [PubMed]
  • Kyhse-Andersen J. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 1984 Dec;10(3-4):203–209. [PubMed]
  • Lam A, Fuller F, Miller J, Kloss J, Manthorpe M, Varon S, Cordell B. Sequence and structural organization of the human gene encoding ciliary neurotrophic factor. Gene. 1991 Jun 30;102(2):271–276. [PubMed]
  • Lehrach H, Diamond D, Wozney JM, Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. [PubMed]
  • Lin LF, Mismer D, Lile JD, Armes LG, Butler ET, 3rd, Vannice JL, Collins F. Purification, cloning, and expression of ciliary neurotrophic factor (CNTF). Science. 1989 Nov 24;246(4933):1023–1025. [PubMed]
  • Lindholm D, Heumann R, Meyer M, Thoenen H. Interleukin-1 regulates synthesis of nerve growth factor in non-neuronal cells of rat sciatic nerve. Nature. 1987 Dec 17;330(6149):658–659. [PubMed]
  • Manthorpe M, Skaper SD, Williams LR, Varon S. Purification of adult rat sciatic nerve ciliary neuronotrophic factor. Brain Res. 1986 Mar 5;367(1-2):282–286. [PubMed]
  • Masiakowski P, Liu HX, Radziejewski C, Lottspeich F, Oberthuer W, Wong V, Lindsay RM, Furth ME, Panayotatos N. Recombinant human and rat ciliary neurotrophic factors. J Neurochem. 1991 Sep;57(3):1003–1012. [PubMed]
  • Millaruelo AI, Nieto-Sampedro M, Yu J, Cotman CW. Neurotrophic activity in the central and peripheral nervous systems of the cat. Effects of injury. Brain Res. 1986 May 21;374(1):12–20. [PubMed]
  • Negro A, Corona G, Bigon E, Martini I, Grandi C, Skaper SD, Callegaro L. Synthesis, purification, and characterization of human ciliary neuronotrophic factor from E. coli. J Neurosci Res. 1991 Jun;29(2):251–260. [PubMed]
  • Raivich G, Kreutzberg GW. Expression of growth factor receptors in injured nervous tissue. I. Axotomy leads to a shift in the cellular distribution of specific beta-nerve growth factor binding in the injured and regenerating PNS. J Neurocytol. 1987 Oct;16(5):689–700. [PubMed]
  • Raivich G, Hellweg R, Kreutzberg GW. NGF receptor-mediated reduction in axonal NGF uptake and retrograde transport following sciatic nerve injury and during regeneration. Neuron. 1991 Jul;7(1):151–164. [PubMed]
  • Rende M, Muir D, Ruoslahti E, Hagg T, Varon S, Manthorpe M. Immunolocalization of ciliary neuronotrophic factor in adult rat sciatic nerve. Glia. 1992;5(1):25–32. [PubMed]
  • Richardson PM, Ebendal T. Nerve growth activities in rat peripheral nerve. Brain Res. 1982 Aug 19;246(1):57–64. [PubMed]
  • Sendtner M, Kreutzberg GW, Thoenen H. Ciliary neurotrophic factor prevents the degeneration of motor neurons after axotomy. Nature. 1990 May 31;345(6274):440–441. [PubMed]
  • Sendtner M, Arakawa Y, Stöckli KA, Kreutzberg GW, Thoenen H. Effect of ciliary neurotrophic factor (CNTF) on motoneuron survival. J Cell Sci Suppl. 1991;15:103–109. [PubMed]
  • Stöckli KA, Lottspeich F, Sendtner M, Masiakowski P, Carroll P, Götz R, Lindholm D, Thoenen H. Molecular cloning, expression and regional distribution of rat ciliary neurotrophic factor. Nature. 1989 Dec 21;342(6252):920–923. [PubMed]
  • Stöckli KA, Lillien LE, Näher-Noé M, Breitfeld G, Hughes RA, Raff MC, Thoenen H, Sendtner M. Regional distribution, developmental changes, and cellular localization of CNTF-mRNA and protein in the rat brain. J Cell Biol. 1991 Oct;115(2):447–459. [PMC free article] [PubMed]
  • Taniuchi M, Clark HB, Johnson EM., Jr Induction of nerve growth factor receptor in Schwann cells after axotomy. Proc Natl Acad Sci U S A. 1986 Jun;83(11):4094–4098. [PMC free article] [PubMed]
  • Thoenen H. The changing scene of neurotrophic factors. Trends Neurosci. 1991 May;14(5):165–170. [PubMed]
  • Unsicker K, Reichert-Preibsch H, Schmidt R, Pettmann B, Labourdette G, Sensenbrenner M. Astroglial and fibroblast growth factors have neurotrophic functions for cultured peripheral and central nervous system neurons. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5459–5463. [PMC free article] [PubMed]
  • Watters DJ, Hendry IA. Purification of a ciliary neurotrophic factor from bovine heart. J Neurochem. 1987 Sep;49(3):705–713. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

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

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...