Cell and Tissue Research

, Volume 320, Issue 3, pp 487–499 | Cite as

Synthesis of transthyretin by the ependymal cells of the subcommissural organ

  • H. A. Montecinos
  • H. Richter
  • T. Caprile
  • E. M. RodríguezEmail author
Regular Article


Transthyretin (TTR) is a protein involved in the transport of thyroid hormones in blood and cerebrospinal fluid (CSF). The only known source of brain-produced TTR is the choroid plexus. In the present investigation, we have identified the subcommissural organ (SCO) as a new source of brain TTR. The SCO is an ependymal gland that secretes glycoproteins into the CSF, where they aggregate to form Reissner’s fibre (RF). Evidence exists that the SCO also secretes proteins that remain soluble in the CSF. To investigate the CSF-soluble compounds secreted by the SCO further, antibodies were raised against polypeptides partially purified from fetal bovine CSF. One of these antibodies (against a 14-kDa compound) reacted with secretory granules in cells of fetal and adult bovine SCO, organ-cultured bovine SCO and the choroid plexus of several mammalian species but not with RF. Western blot analyses with this antibody revealed two polypeptides of 14 kDa and 40 kDa in the bovine SCO, in the conditioned medium of SCO explants, and in fetal and adult bovine CSF. Since the monomeric and tetrameric forms of TTR migrate as bands of 14 kDa and 40 kDa by SDS-polyacrylamide gel electrophoresis, a commercial preparation of human TTR was run, with both bands being reactive with this antibody. Bovine SCO was also shown to synthesise mRNA encoding TTR under in vivo and in vitro conditions. We conclude that the SCO synthesises TTR and secretes it into the CSF. Colocalisation studies demonstrated that the SCO possessed two populations of secretory cells, one secreting both RF glycoproteins and TTR and the other secreting only the former. TTR was also detected in the SCO of bovine embryos suggesting that this ependymal gland is an important source of TTR during brain development.


Subcommissural organ Choroid plexus Transthyretin Cerebrospinal fluid Bovine 


  1. Achen MG, Harms PJ, Thomas T, Richardson SJ, Wettenhall REH, Schreiber G (1992) Protein synthesis at the blood-brain barrier: the major protein secreted by amphibian choroid plexus is a lipocalin. J Biol Chem 267:23170–23174Google Scholar
  2. Achen MG, Duan W, Petterson TM, Harms PJ, Richardson SJ, Lawrence MC, Wettenhall REH, Aldred AR, Schreiber G (1993) Transthyretin gene expression in choroid plexus first evolved in reptiles. Am J Physiol 265:R982–R989Google Scholar
  3. Anderson GW (2001) Thyroid hormones and the brain. Front Neuroendocrinol 22:1–17Google Scholar
  4. Bellovino D, Moritomo T, Pisaniello A, Gaetani S (1998) In vitro and in vivo studies on transthyretin oligomerization. Exp Cell Res 243:101–112Google Scholar
  5. Blake C, Geisow M, Oatley S, Rerat B, Rerat C (1978) Structure of prealbumin: secondary, tertiary and quaternary interactions determined by Fourier refinement at 1,8. J Mol Biol 121:339–356Google Scholar
  6. Cozzani C, Hartmann B (1980) Preparation of antibodies specific to choline acetyltransferase from bovine caudate nucleus and immunohistochemical localization of the enzyme. Proc Natl Acad Sci USA 77:7453–7457Google Scholar
  7. Dickson PW, Aldred AR, Maarley PD, Bannister D, Schreiber G (1986) Rat choroid plexus specializes in the synthesis and secretion of transthyretin (prealbumin)—regulation of transthyretin synthesis in choroid plexus in independent from that in liver. J Biol Chem 261:3475–3478Google Scholar
  8. Divino CM, Schussler GC (1990) Receptor-mediated uptake and internalization of transthyretin. J Biol Chem 265:1425–1429Google Scholar
  9. Dussault JH, Ruel J (1987) Thyroid hormone and brain development. Annu Rev Physiol 49:321–334Google Scholar
  10. Episkopou V, Maeda S, Nishiguchi S, Shimada K, Gaitanaris GA, Gottesman ME, Robertson JE (1993) Disruption of the transthyretin gene results in mice with depressed level of plasma retinol and thyroid hormone. Proc Natl Acad Sci USA 90:2375–2379Google Scholar
  11. Fernández-Llebrez P, Pérez-Figares JM, Becerra J, Perez J, Marín-Girón F (1984) Morphological evidence for the presence of two cell types in the ependyma of the subcommissural organ of the snake, Natrix maura. Cell Tissue Res 238:407–409Google Scholar
  12. Fung W, Thomas T, Dickson P, Aldred A, Milland J, Dziadek M, Power B, Hudson P, Schreiber G (1988) Structure and expression of the rat transthyretin (prealbumin) gene. J Biol Chem 263:480–488Google Scholar
  13. González CB, Rodríguez EM (1980) Ultrastructure and immunocytochemistry of neurons in the supraoptic and paraventricular nuclei of the lizard Liolaemus cyanogaster. Evidence for the intracisternal location of the precursor of neurophysin. Cell Tissue Res 207:463–477Google Scholar
  14. Hofer H (1959) Zur Morphologie der circumventriculären Organe des Zwischenhirns der Säugetiere. Verh Dtsch Zool Ges 22:202–251Google Scholar
  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680Google Scholar
  16. Larsen N, Kirkpatrick B (1996) Seven genes from human chromosome 18 map to chromosome 24 in the bovine. Cytogenet Cell Genet 73:184–186Google Scholar
  17. Lauder JM, Bloom FE (1974) Ontogeny of monoamine neurons in the locus coeruleus, raphe nuclei and substantia nigra of the rat. I. Cell differentiation. J Comp Neurol 155:469–481Google Scholar
  18. Mendel CM (1989) The free hormone hypothesis: a physiologically based mathematical model. Endocr Rev 10:232–274Google Scholar
  19. Mendes M, Joao M (2001) Internalization of transthyretin. Evidence of a novel yet unidentified receptor associated protein (RAP)-sensitive receptor. J Biol Chem 276:14420–14425Google Scholar
  20. Meiniel R, Molat JL, Duchier-Liris N, Meiniel A (1990) Ontogenesis of the secretory epithelium of the bovine subcommissural organ. A histofluorescence study using lectins and monoclonal antibodies. Dev Brain Res 55:171–180Google Scholar
  21. Meiniel A, Meiniel R, Didier R, Creveaux I, Gobron S, Monnerie H, Dastugue B (1996) The subcommissural organ and Reissner’s fibre complex—an enigma in the central nervous system? Prog Histochem Cytochem 30:1–66Google Scholar
  22. Nualart F, Hein S, Rodríguez EM, Oksche A (1991) Identification and partial characterization of the secretory glycoproteins of the bovine subcommissural organ Reissner’s fiber complex. Evidence for the existence of two precursor forms. Mol Brain Res 11:227–238Google Scholar
  23. O’Farrel PZ, Goodman HM, O’Farrel PH (1977) High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142Google Scholar
  24. Palha JA, Thompson M, Morreale G, Episkopou V, Gottesman M, Mascarenhas MJ (1997) Transthyretin is not essential for thyroxine to reach the brain and other tissue in transthyretin-null mice. Am J Physiol 272:E485–E493Google Scholar
  25. Pérez-Fígares JM, Jiménez A, Rodríguez EM (2001) Subcommissural organ, cerebrospinal fluid circulation and hydrocephalus. Microsc Res Tech 52:591–697Google Scholar
  26. Peruzzo B, Pérez J, Fernández-Llebrez P, Pérez-Figares JM, Rodríguez EM (1990) Ultrastructural immunocytochemistry and lectin histochemistry of the subcommissural organ in the snake Natrix maura with particular emphasis on its vascular and leptomeningeal projections. Histochemistry 93:269–277Google Scholar
  27. Pettersson T, Carlstrom A, Jornvall H (1987) Different types of microheterogeneity of human thyroxine-binding prealbumin. Biochemistry 26:4572–4583Google Scholar
  28. Pettersson T, Carlstrom A, Ehrenberg A, Jornvall H (1989) Transthyretin microheterogeneity and thyroxine binding are influenced by non-amino acid components and glutathione constituents. Biochem Biophys Res Commun 158:341–347Google Scholar
  29. Pettersson T, Ernstron U, Griffiths W, Sjovall J, Bergman T, Jornvall H (1995) Lutein associated with a transthyretin indicates carotenoid derivation and novel multiplicity of transthyretin ligands. FEBS Lett 365:23–26Google Scholar
  30. Reid DG, MacLachlan LK, Voylo M, Leeson PD (1989) A proton and fluorine-nuclear magnetic resonance and fluorescence study of the binding of some natural and synthetic thyromimetics to prealbumin (transthyretin). J Biol Chem 264:2013–2023Google Scholar
  31. Richter HG, Munoz RI, Millán CS, Guiñazú MF, Yulis CR, Rodríguez EM (2001) The floor plate cells from bovines express the mRNA encoding for SCO-spondin and its translation products. Mol Brain Res 93:137–147Google Scholar
  32. Rodríguez EM, Oksche A, Hein S, Rodríguez S, Yulis CR (1984a) Comparative immunocytochemical study of the subcommissural organ. Cell Tissue Res 237:427–441Google Scholar
  33. Rodríguez EM, Yulis R, Peruzzo B, Alvial G, Andrade R (1984b) Standardization of various applications of methacrylate embedding and silver methenamine for light and electron microscopy immunocytochemistry. Histochemistry 81:253–263Google Scholar
  34. Rodríguez EM, Garrido O, Oksche A (1990) Lectin histochemistry of the human fetal subcommissural organ. Cell Tissue Res 262:105–113Google Scholar
  35. Rodríguez EM, Oksche A, Hein S, Yulis CR (1992) Cell biology of the subcommissural organ. Int Rev Cytol 135:39–121Google Scholar
  36. Rodríguez EM, Jara P, Richter H, Montecinos H, Flandes B, Wiegand R, Oksche A (1993) Evidence of the release of CSF-soluble secretory material from the subcommissural organ, with particular reference to the situation in human. In: Oksche A, Rodríguez EM, Fernández-Llebrez P (eds) The subcommissural organ: an ependymal brain gland. Springer, Berlin Heidelberg New York, pp 121–131Google Scholar
  37. Rodríguez EM, Rodríguez S, Hein S (1998) The subcommissural organ. Microsc Res Tech 41:98–123Google Scholar
  38. Rodríguez EM, Oksche A, Montecinos H (2001) Human subcommissural organ, with particular emphasis on its secretory activity during the fetal life. Microsc Res Tech 52:573–590Google Scholar
  39. Schoebitz K, Garrido O, Heinrichs M, Speer L, Rodríguez EM (1986) Ontogenetic development of the chick and duck subcommissural organ. An immunocytochemical study. Histochemistry 84:31–40Google Scholar
  40. Schoebitz K, Rodríguez EM, Garrido O (1993) Ontogenetic development of the subcommissural organ with reference to the flexural organ. In: Oksche A, Rodríguez EM, Fernández-Llebrez P (eds) The subcommissural organ: an ependymal brain gland. Springer, Berlin Heidelberg New York, pp 41–49Google Scholar
  41. Schoebitz K, González C, Peruzzo B, Yulis CR, Rodríguez EM (2001) Organ culture of the bovine subcommissural organ. Evidence for synthesis and release of the secretory material. Microsc Res Tech 52:496–509Google Scholar
  42. Schreiber G (2002) The evolutionary and integrative roles of transthyretin in thyroid hormone homeostasis. J Endocrinol 175:61–73Google Scholar
  43. Soprano DR, Herbert J, Soprano KJ, Schon EA, Goodman DS (1985) Demonstration of transthyretin mRNA in the brain and other extrahepatic tissue in the rat. J Biol Chem 260:11793–11798Google Scholar
  44. Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry. Preparation and properties of soluble antigen–antibody complex (horseradish-peroxidase–antiperoxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333Google Scholar
  45. Thomas T, Power B, Hudson P, Schreiber G, Dziadek M (1988) The expression of transthyretin mRNA in the developing rat brain. Dev Biol 128:415–427Google Scholar
  46. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheet: procedure and some applications. Proc Natl Acad Sci USA 53:4350–4354Google Scholar
  47. Walker GR, Feather KD, Davis PD, Hines KK (1995) Super Signal TM CL-HRP: a new enhanced chemiluminescent substrate for the development of the horseradish peroxide label in Western blotting applications. J NIH Res 7:76Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • H. A. Montecinos
    • 1
    • 2
  • H. Richter
    • 1
  • T. Caprile
    • 1
    • 2
  • E. M. Rodríguez
    • 1
    Email author
  1. 1.Facultad de Medicina, Instituto de Histología y PatologíaUniversidad Austral de ChileValdiviaChile
  2. 2.Departamento de Biología Celular, Facultad de Ciencias BiológicasUniversidad de ConcepciónConcepciónChile

Personalised recommendations