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. 1991 Oct 2;115(2):447–459. doi: 10.1083/jcb.115.2.447

Regional distribution, developmental changes, and cellular localization of CNTF-mRNA and protein in the rat brain

PMCID: PMC2289163  PMID: 1918150

Abstract

Ciliary neurotrophic factor (CNTF) is a potent survival molecule for a variety of embryonic neurons in culture. The developmental expression of CNTF occurs clearly after the time period of the physiological cell death of CNTF-responsive neurons. This, together with the sites of expression, excludes CNTF as a target-derived neuronal survival factor, at least in rodents. However, CNTF also participates in the induction of type 2 astrocyte differentiation in vitro. Here we demonstrate that the time course of the expression of CNTF-mRNA and protein in the rat optic nerve (as evaluated by quantitative Northern blot analysis and biological activity, respectively) is compatible with such a glial differentiation function of CNTF in vivo. We also show that the type 2 astrocyte-inducing activity previously demonstrated in optic nerve extract can be precipitated by an antiserum against CNTF. Immunohistochemical analysis of astrocytes in vitro and in vivo demonstrates that the expression of CNTF is confined to a subpopulation of type 1 astrocytes. The olfactory bulb of adult rats has comparably high levels of CNTF to the optic nerve, and here again, CNTF- immunoreactivity is localized in a subpopulation of astrocytes. However, the postnatal expression of CNTF in the olfactory bulb occurs later than in the optic nerve. In other brain regions both CNTF-mRNA and protein levels are much lower.

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Selected References

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  1. Adler R., Landa K. B., Manthorpe M., Varon S. Cholinergic neuronotrophic factors: intraocular distribution of trophic activity for ciliary neurons. Science. 1979 Jun 29;204(4400):1434–1436. doi: 10.1126/science.451576. [DOI] [PubMed] [Google Scholar]
  2. 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. doi: 10.1523/JNEUROSCI.10-11-03507.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barbin G., Manthorpe M., Varon S. Purification of the chick eye ciliary neuronotrophic factor. J Neurochem. 1984 Nov;43(5):1468–1478. doi: 10.1111/j.1471-4159.1984.tb05410.x. [DOI] [PubMed] [Google Scholar]
  4. Belin D., Wohlwend A., Schleuning W. D., Kruithof E. K., Vassalli J. D. Facultative polypeptide translocation allows a single mRNA to encode the secreted and cytosolic forms of plasminogen activators inhibitor 2. EMBO J. 1989 Nov;8(11):3287–3294. doi: 10.1002/j.1460-2075.1989.tb08489.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. 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. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  6. 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. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Doucette J. R., Kiernan J. A., Flumerfelt B. A. The re-innervation of olfactory glomeruli following transection of primary olfactory axons in the central or peripheral nervous system. J Anat. 1983 Aug;137(Pt 1):1–19. [PMC free article] [PubMed] [Google Scholar]
  8. Ebendal T., Jacobson C. O. Tissue explants affecting extension and orientation of axons in cultured chick embryo ganglia. Exp Cell Res. 1977 Mar 15;105(2):379–387. doi: 10.1016/0014-4827(77)90135-5. [DOI] [PubMed] [Google Scholar]
  9. Ebendal T. Stage-dependent stimulation of neurite outgrowth exerted by nerve growth factor and chick heart in cultured embryonic ganglia. Dev Biol. 1979 Oct;72(2):276–290. doi: 10.1016/0012-1606(79)90118-0. [DOI] [PubMed] [Google Scholar]
  10. Ernsberger U., Sendtner M., Rohrer H. Proliferation and differentiation of embryonic chick sympathetic neurons: effects of ciliary neurotrophic factor. Neuron. 1989 Mar;2(3):1275–1284. doi: 10.1016/0896-6273(89)90312-7. [DOI] [PubMed] [Google Scholar]
  11. Hatten M. E., Lynch M., Rydel R. E., Sanchez J., Joseph-Silverstein J., Moscatelli D., Rifkin D. B. In vitro neurite extension by granule neurons is dependent upon astroglial-derived fibroblast growth factor. Dev Biol. 1988 Feb;125(2):280–289. doi: 10.1016/0012-1606(88)90211-4. [DOI] [PubMed] [Google Scholar]
  12. Helfand S. L., Smith G. A., Wessells N. K. Survival and development in culture of dissociated parasympathetic neurons from ciliary ganglia. Dev Biol. 1976 Jun;50(2):541–547. doi: 10.1016/0012-1606(76)90174-3. [DOI] [PubMed] [Google Scholar]
  13. 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] [Google Scholar]
  14. Hill M. A., Bennett M. R. Cholinergic growth factor from skeletal muscle elevated following denervation. Neurosci Lett. 1983 Jan 31;35(1):31–35. doi: 10.1016/0304-3940(83)90522-0. [DOI] [PubMed] [Google Scholar]
  15. Hofmann H. D. Ciliary neuronotrophic factor stimulates choline acetyltransferase activity in cultured chicken retina neurons. J Neurochem. 1988 Jul;51(1):109–113. doi: 10.1111/j.1471-4159.1988.tb04842.x. [DOI] [PubMed] [Google Scholar]
  16. Hofmann H. D. Development of cholinergic retinal neurons from embryonic chicken in monolayer cultures: stimulation by glial cell-derived factors. J Neurosci. 1988 Apr;8(4):1361–1369. doi: 10.1523/JNEUROSCI.08-04-01361.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hughes S. M., Lillien L. E., Raff M. C., Rohrer H., Sendtner M. Ciliary neurotrophic factor induces type-2 astrocyte differentiation in culture. Nature. 1988 Sep 1;335(6185):70–73. doi: 10.1038/335070a0. [DOI] [PubMed] [Google Scholar]
  18. Johnson J. E., Barde Y. A., Schwab M., Thoenen H. Brain-derived neurotrophic factor supports the survival of cultured rat retinal ganglion cells. J Neurosci. 1986 Oct;6(10):3031–3038. doi: 10.1523/JNEUROSCI.06-10-03031.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kitagawa T., Aikawa T. Enzyme coupled immunoassay of insulin using a novel coupling reagent. J Biochem. 1976 Jan;79(1):233–236. doi: 10.1093/oxfordjournals.jbchem.a131053. [DOI] [PubMed] [Google Scholar]
  20. Kostura M. J., Tocci M. J., Limjuco G., Chin J., Cameron P., Hillman A. G., Chartrain N. A., Schmidt J. A. Identification of a monocyte specific pre-interleukin 1 beta convertase activity. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5227–5231. doi: 10.1073/pnas.86.14.5227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  22. Lehwalder D., Jeffrey P. L., Unsicker K. Survival of purified embryonic chick retinal ganglion cells in the presence of neurotrophic factors. J Neurosci Res. 1989 Oct;24(2):329–337. doi: 10.1002/jnr.490240225. [DOI] [PubMed] [Google Scholar]
  23. Lillien L. E., Raff M. C. Differentiation signals in the CNS: type-2 astrocyte development in vitro as a model system. Neuron. 1990 Aug;5(2):111–119. doi: 10.1016/0896-6273(90)90301-u. [DOI] [PubMed] [Google Scholar]
  24. Lillien L. E., Sendtner M., Raff M. C. Extracellular matrix-associated molecules collaborate with ciliary neurotrophic factor to induce type-2 astrocyte development. J Cell Biol. 1990 Aug;111(2):635–644. doi: 10.1083/jcb.111.2.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lillien L. E., Sendtner M., Rohrer H., Hughes S. M., Raff M. C. Type-2 astrocyte development in rat brain cultures is initiated by a CNTF-like protein produced by type-1 astrocytes. Neuron. 1988 Aug;1(6):485–494. doi: 10.1016/0896-6273(88)90179-1. [DOI] [PubMed] [Google Scholar]
  26. Lin L. F., Mismer D., Lile J. D., Armes L. G., Butler E. T., 3rd, Vannice J. L., Collins F. Purification, cloning, and expression of ciliary neurotrophic factor (CNTF). Science. 1989 Nov 24;246(4933):1023–1025. doi: 10.1126/science.2587985. [DOI] [PubMed] [Google Scholar]
  27. Liu F. T., Zinnecker M., Hamaoka T., Katz D. H. New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates. Biochemistry. 1979 Feb 20;18(4):690–693. doi: 10.1021/bi00571a022. [DOI] [PubMed] [Google Scholar]
  28. McCarthy K. D., de Vellis J. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol. 1980 Jun;85(3):890–902. doi: 10.1083/jcb.85.3.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McManaman J., Crawford F., Clark R., Richker J., Fuller F. Multiple neurotrophic factors from skeletal muscle: demonstration of effects of basic fibroblast growth factor and comparisons with the 22-kilodalton choline acetyltransferase development factor. J Neurochem. 1989 Dec;53(6):1763–1771. doi: 10.1111/j.1471-4159.1989.tb09241.x. [DOI] [PubMed] [Google Scholar]
  30. Miller R. H., David S., Patel R., Abney E. R., Raff M. C. A quantitative immunohistochemical study of macroglial cell development in the rat optic nerve: in vivo evidence for two distinct astrocyte lineages. Dev Biol. 1985 Sep;111(1):35–41. doi: 10.1016/0012-1606(85)90432-4. [DOI] [PubMed] [Google Scholar]
  31. Nieto-Sampedro M., Lewis E. R., Cotman C. W., Manthorpe M., Skaper S. D., Barbin G., Longo F. M., Varon S. Brain injury causes a time-dependent increase in neuronotrophic activity at the lesion site. Science. 1982 Aug 27;217(4562):860–861. doi: 10.1126/science.7100931. [DOI] [PubMed] [Google Scholar]
  32. Raisman G. Specialized neuroglial arrangement may explain the capacity of vomeronasal axons to reinnervate central neurons. Neuroscience. 1985 Jan;14(1):237–254. doi: 10.1016/0306-4522(85)90176-9. [DOI] [PubMed] [Google Scholar]
  33. Rubartelli A., Cozzolino F., Talio M., Sitia R. A novel secretory pathway for interleukin-1 beta, a protein lacking a signal sequence. EMBO J. 1990 May;9(5):1503–1510. doi: 10.1002/j.1460-2075.1990.tb08268.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Saadat S., Sendtner M., Rohrer H. Ciliary neurotrophic factor induces cholinergic differentiation of rat sympathetic neurons in culture. J Cell Biol. 1989 May;108(5):1807–1816. doi: 10.1083/jcb.108.5.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sendtner M., Kreutzberg G. W., Thoenen H. Ciliary neurotrophic factor prevents the degeneration of motor neurons after axotomy. Nature. 1990 May 31;345(6274):440–441. doi: 10.1038/345440a0. [DOI] [PubMed] [Google Scholar]
  36. Spranger Matthias, Lindholm Dan, Bandtlow Christine, Heumann Rolf, Gnahn Hannes, Näher-Noé Martina, Thoenen Hans. Regulation of Nerve Growth Factor (NGF) Synthesis in the Rat Central Nervous System: Comparison between the Effects of Interleukin-1 and Various Growth Factors in Astrocyte Cultures and in vivo. Eur J Neurosci. 1990 Jan;2(1):69–76. doi: 10.1111/j.1460-9568.1990.tb00382.x. [DOI] [PubMed] [Google Scholar]
  37. Stöckli K. A., 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. doi: 10.1038/342920a0. [DOI] [PubMed] [Google Scholar]
  38. 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. doi: 10.1073/pnas.84.15.5459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Watters D. J., Hendry I. A. Purification of a ciliary neurotrophic factor from bovine heart. J Neurochem. 1987 Sep;49(3):705–713. doi: 10.1111/j.1471-4159.1987.tb00951.x. [DOI] [PubMed] [Google Scholar]

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