Abstract
Tissue of the secretory, glial subcommissural organ (SCO) of adult, male cattle was cultured in serum-free medium for 70 days in vitro. Only minor alterations in the histoarchitecture and the cytology of the explanted SCOs could be observed by light and electron microscopy. Light- and electron-microscopic immunocytochemical investigations with an antiserum raised against bovine SCO secretory proteins revealed intra- and extra-cellularly localized immunoreactive material in tissue sections of SCO explants cultured up to 69 days in vitro. An indirect competition enzyme-linked immunosorbent assay (ELISA) was developed to detect minor quantities of SCO secretory products. By means of this assay, approximately 35 ng RF protein per ml was detected in culture medium supernatants conditioned for 3 days in SCO tissue cultures at 3, 38, and 69 days in vitro. These studies demonstrate that the bovine SCO can maintain its secretory activity throughout long periods in vitro.
Similar content being viewed by others
References
Bank H (1988) A quantitative enzyme-linked immunosorbent assay for rat insulin. J Immunoassay 9:135–158
Barlow RM, D'Agostino AN, Cancilla PA (1966) A morphological and histochemical study of the subcommissural organ of young and old sheep. Z Zellforsch Mikrosk Anat 77:299–315
Benda P, De Vitry F, Picart R, Tixier-Vidal A (1975) Dissociated cell cultures from fetal mouse hypothalamus. Patterns of organization and ultrastructural features. Exp Brain Res 23:29–47
Bruel MT, Meiniel R, Meiniel A, David D (1987) Ontogenetical study of the chick embryo subcommissural organ by lectin-histofluorescence and electron microscopy. J Neural Transm 70:145–168
Butler JE (1982) The amplified ELISA: principles of an application for the comparative quantitation of class and subclass of antibodies and the distribution of antibodies and antigens in bio-chemical separates. In: Langone JJ, Van Vunakis H (eds) Methods in Enzymology, vol 73. Academic Press, New York, pp 482–453
Chen ILI, Lu KS, Lin HS (1973) Electron microscopic and cytochemical studies of the mouse subcommissural organ. Z Zellforsch Mikrosk Anat 139:217–236
De Mey J (1983) Colloidal gold probes in immunocytochemistry. In: Polak M, Van Norder S (eds) Immunocytochemistry. Practical application in pathology and biology. Wright, Bristol London, pp 217–236
Didier-Bazes M, Chouaf L, Hardin H, Aguera M, Fèvre-Montagne M, Belin MF (1991) Developmental neuron-glia interaction: role of the serotonin innervation upon the onset of GABA uptake into the ependymocytes of the rat subcommissural organ. Dev Brain Res 63:135–139
Engvall E, Perlmann P (1972) Enzyme-linked immunosorbent assay, ELISA. III. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol 109:129–135
Gabrion J, Peraldi S, Faivre-Baumann A, Klotz C, Ghandour MS, Paulin D, Assenmacher I, Tixier-Vidal A (1988) Characterization of ependymal cells in hypothalamic and choroideal primary culture. Neuroscience 24:993–1007
Herrlinger H (1970) Licht- und elektronenmikroskopische Untersuchungen am Subcommissuralorgan der Maus. Ergeb Anat Entwicklungsgesch 42:1–73
Hild W (1957) Ependymal cells in culture. Z Zellforsch Mikrosk Anat 46:259–271
Hofer H (1959) Zur Morphologie der circumventrikulären Organe des Zwischenhirnes der Säugetiere. Zool Anz [Suppl] 22:251–264
Hubbuch A, Debus E, Linke R, Schrenk WJ (1986) Enzyme-immunoassay: a review. Prog Clin Biochem 4:109–146
Isomäki AM, Kivalo E, Talanti S (1965) Electron-microscopic structure of the subcommissural organ in the calf (Bos taurus) with special reference to the secretory phenomena. Ann Acad Sci Fenn (Med) 111:1–64
Karoumi A, Croisille Y, Croisille F, Meiniel R, Belin MF, Meiniel A (1990) Glycoprotein synthesis in the subcommissural organ of the chick embryo. II. An immunochemical study. J Neural Transm [GenSect] 80:203–212
Karoumi A, Meiniel R, Belin MF, Meiniel A (1991) A comparative immunocytochemical and immunochemical analysis of the glycoproteins synthesized in the bovine subcommissural organ. J Neural Transm [GenSect] 86:205–216
Krstic RV (1976) Ultrastruktur der Säugetierzelle. Ein Atlas zum Studium für Mediziner und Biologen. Springer, Berlin Heidelberg New York, pp 1–377
Lehmann W, Sterba G (1989) Tissue culture of rat subcommissural organ in vitro. Biomed Res [Suppl 3] 10:11–18
Lehmann W, Sterba G, Wobus AM (1989) Primary culture of the mouse subcommissural organ in vitro. Acta Zool (Stockh) 70:199–203
Leonhardt H (1980) Ependym und circumventriculäre Organe. In: Oksche A, Vollrath L (eds) Handbuch der mikroskopischen Anatomie des Menschen, Band 4, Teil 10, Neuroglia I. Springer, Berlin Heidelberg New York, pp 177–666
Lindberg JA, Talanti S (1975) The surface fine structure of the bovine subcommissural organ. Cell Tissue Res 163:125–132
Lösecke W, Naumann W, Sterba G (1984) Preparation and discharge of secretion in the subcommissural organ in the rat. Electron-microscopic immunocytochemical study. Cell Tissue Res 235:201–206
Lösecke W, Naumann W, Sterba G (1986) Immuno-electron-microscopic analysis of the basal route of secretion in the subcommissural organ of the rabbit. Cell Tissue Res 244:449–456
Meiniel A, Molat JL, Meiniel R (1988) Complex-type glycoproteins synthesized in the subcommissural organ of mammals. Light- and electron-microscopic Einvestigations by use of lectins. Cell Tissue Res 253:383–395
Meiniel A, Meiniel R, Karoumi A, Duchier-Liris N, Molat JL (1992) The bovine subcommissural organ: cytochemical and immunochemical characterization of the secretory process. Prog Brain Res 91:331–342
Meiniel R, Molat JL, Duchier Liris N, Meiniel A (1990) Ontogenesis of the secretory epithelium of the bovine subcommissural organ. A histofiuorescence study using lectins and monoclonal antibodies. Brain Res Dev Brain Res 55:171–180
Meiniel R, Duchier-Liris N, Molat J-L, Meiniel A (1991) The complex-type glycoprotein secreted by the bovine subcommissural organ: an immunological study using C1B8A8 monoclonal antibody. Cell Tissue Res 266:483–490
Naumann W, Lehmann W, Debbage P (1993) The subcommissural organ and ontogenetic development of the brain. In: Oksche A (ed) The subcommissural organ. Springer, Berlin Heidelberg New York, pp 61–69
Nualart F, Hein S, Rodriguez EM, Oksche A (1991) Identification and partial characterization of the secretory glycoproteins of the subcommissural organ — Reissner's fiber complex. Evidence for the existence of two precursor forms. Mol Brain Res 11:227–238
Oksche A (1961) Vergleichende Untersuchungen über die sekretorische Aktivität des Subkommissuralorgans und den Gliacharakter seiner Zellen. Z Zellforsch Mikrosk Anat 54:549–612
Olsson R (1958) Studies on the subcommissural organ. Acta Zool (Stockh) 39:71–102
Raff MC, Abney ER, Cohen J, Lindsay R, Noble M (1983) Two types of astrocytes in cultures of developing rat white matter: differences in morphology, surface gangliosides and growth characteristics. J Neurosci 3:1289–1300
Reid LM, Rojkind M (1979) New techniques for culturing differentiated cells: reconstituted basement membrane rafts. In: Jakoby WB, Pastan IH (eds) Methods in enzymology, vol 58. Academic Press, New York, pp 263–278
Reisert I, Lang R, Pilgrim C (1980) Differenzierung von Glia- und Nervenzellen in Primärkultur aus dem Hypothalamus der Ratte. Verh Anat Ges 74:677–683
Rieke GK, Jordan FL, Wynder HJ, Thomas WE (1987) Ultrastructure of ependymal cells in primary cultures of cerebral cortex. J Neurosci Res 18:484–492
Rodriguez EM (1970) Ependymal specializations. II. Ultrastructural aspects of the apical secretion of the toad subcommissural organ. Z Zellforsch Mikrosk Anat 111:15–31
Rodriguez EM, Oksche A, Hein S, Rodriguez S, Yulis R (1984) Comparative immunocytochemical study of the subcommissural organ. Cell Tissue Res 237:427–441
Rodriguez EM, Herrera H, Peruzzo B, Rodriguez S, Hein S, Oksche A (1986) Light- and electron-microscopic immunocytochemistry and lectin histochemistry of the subcommissural organ: Evidence for processing of the secretory material. Cell Tissue Res 243:545–559
Rodriguez EM, Hein S, Rodriguez S, Herrera H, Peruzzo B, Nualart F, Oksche A (1987a) Analysis of the secretory products of the subcommissural organ. In: Scharrer B, Korf HW, Hartwig HG (eds) Functional morphology of the neuroendocrine systems. Evolutionary and environmental aspects. Springer, Berlin Heidelberg New York, pp 189–202
Rodriguez EM, Oksche A, Rodriguez S, Hein S, Peruzzo B, Schoebitz K, Herrera H (1987b) The subcommissural organ and Reissner's fibre. Fine structure and cytochemistry. In: Gross PM (ed) Circumventricular organs and body fluids. CRC Press, Boca Raton, pp 3–41
Rodriguez EM, Rodriguez S, Schoebitz K, Yulis CR, Hoffmann P, Manns V, Oksche A (1989) Light- and electron-microscopic investigation of the rat subcommissural organ grafted under the kidney capsule, with particular reference to immunocytochemistry and lectin histochemistry. Cell Tissue Res 258:499–514
Rühle HJ (1971) Anomalien im Wachstum der Achsenorgane nach experimenteller Ausschaltung des Komplexes Subcommissu-ralorgan-Reissnerscher Faden. Untersuchungen am Rippenmolch (Pleurodeles waltli MICHAH. 1830). Acta Zool (Stockh) 52:23–68
Schneider B (1972) Ventrikelseitige Beeinflussung des Subcommissuralorgans der Ratte mit hyperosmotischen Lösungen. Dissertation A; Karl-Marx-Universität Leipzig
Stanka P, Schwink A, Wetzstein R (1964) Elektronenmikroskopische Untersuchung des Subcommissuralorgans der Ratte. Z Zellforsch Mikrosk Anat 63:277–301
Sterba G, Kießig C, Kleim I, Naumann W, Petter H (1982a) Immunzytochemische Untersuchungen an den Subkommissuralorganen von Rind, Ziege und Schaf. Biol Zbl 101:241–248
Sterba G, Kießig C, Naumann W, Petter H, Kleim I (1982b) The secretion of the subcommissural organ. A comparative immunocytochemical investigation. Cell Tissue Res 226:427–439
Sterba G, Kleim I, Naumann W, Petter H (1981) Immunocytochemical investigation of the subcommissural organ in the rat. Cell Tissue Res 218:659–662
Sternberger LA, Hardy PH, 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 18:315–333
Talanti S (1958) Studies on the subcommissural organ in some domestic animals. Ann Med Exp Fenn [Suppl 9] 36:1–97
Turkewitsch N (1936) Die Entwicklung des subkommissuralen Organs beim Rind (Bos taurus L.). Morphol Jahrb 77:573–586
Voller A, Bartlett A, Bidwell DE (1978) Enzyme immunoassays with special reference to ELISA techniques. J Clin Pathol 31:507–520
Wolf G, Sterba G (1972) Zur stofflichen Charakteristik des Reissnerschen Fadens. Acta Zool (Stockh) 53:147–154
Yorde DE, Sasse EA, Wang TY, Hussa RO, Garancis JC (1976) Competitive enzyme-linked immunoassay with use of soluble enzyme/antibody immune complexes for labeling. I. Measurement of human choriogonadotropin. Clin Chem 22:1372–1377
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lehmann, W., Naumann, W. & Wagner, U. Tissue culture of bovine subcommissural organ. Anat Embryol 187, 505–514 (1993). https://doi.org/10.1007/BF00174426
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00174426