The secretion of the subcommissural organ

Sterba, G.; Kiessig, C; Naumann, W.; Petter, H; Kleim, I

doi:10.1007/bf00218370

Cited by 80 publications

(11 citation statements)

References 4 publications

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“…A major difficulty has been the high degree of packaging and insolubility of RF-glycoproteins. Electrophoretic analyses under denaturing and reducing conditions suggest that RF is composed of polypeptides with a large range of molecular masses (10-450 kDa; Sterba et al 1987;Nualart et al 1991). The analysis of the compositions of RF-glycoproteins has shown that they are rich in cysteine and in hydrophobic amino acids (Hädge and Sterba 1973;Rodríguez et al 1992).…”

Section: Introductionmentioning

confidence: 99%

Immunochemical analysis of the subcommissural organ-Reissner's fiber complex using antibodies against alkylated and deglycosylated glycoproteins of the bovine Reissner's fiber

Nualart

Rodríguez

1996

Cell and Tissue Research

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Reissner's fiber (RF) has been isolated, solubilized, and used to raise polyclonal antibodies. The present investigation has been designed: (1) to obtain antibodies against RF-glycoproteins in their native form (anti-RF-BI), after irreversible denaturation by alkylation (anti-RF-A), and after alkylation and deglycosylation by using endoglycosidase F (anti-RF-DE); (2) to use these antisera for a comparative immunocytochemical study of the subcommissural organ (SCO)-RF complex; (3) to establish the molecular mass of the deglycosylated RF-glycoproteins. Anti-RF-BI reacts with the SCO of all the species investigated. Anti-RF-A and anti-RF-DE only react with bovine SCO and RF. The three antisera stain the same bands in immunoblots of extracts of bovine SCO and RF, but anti-RF-A and anti-RF-DE reveal additional bands. The epitope common to all species reacting with anti-RF-BI is thus probably conformational in nature and associated with the integrity of the disulfide bonds. The lack of antibodies against conserved sequential epitopes in anti-RF-A does not support previous assumptions on the conserved nature of the SCO secretion. After deglycosylation by using endoglycosidase F, the RF-glycoproteins present a reduction in their molecular mass ranging between 10% and 25%. The three larger compounds (450, 300, and 230 kDa) lose their affinity for Limax flavus agglutinin (affinity = sialic acid), whereas the 190-kDa compound (170 kDa after deglycosylation) keeps its affinity for this lectin suggesting that it has N-linked and O-linked carbohydrate moieties, the three larger proteins probably having only N-linked carbohydrates.

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Section: Introductionmentioning

confidence: 99%

Immunochemical analysis of the subcommissural organ-Reissner's fiber complex using antibodies against alkylated and deglycosylated glycoproteins of the bovine Reissner's fiber

Nualart

Rodríguez

1996

Cell and Tissue Research

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“…In Reissner's fiber of the bovine numerous polypeptides were identified after SDS-PAGE (40 bands ranging from 10 to 350kD; Sterba et al, 1987). Only one polypeptide (32 kD) was found after immunoaffinity chromatography of soluble extracts of the medulla oblongata in the chick embryo.…”

Section: Discussionmentioning

confidence: 98%

“…Karoumi et al than 40 polypeptides ranging from 10 to 350kD could be identified. Under non-denaturating conditions (Sterba et al, 1987) only two bands were revealed.…”

Section: Introductionmentioning

confidence: 93%

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Glycoprotein synthesis in the subcommissural organ of the chick embryo II. an immunochemical study

Karoumi

Croisille

et al. 1990

J. Neural Transmission

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In the chick embryo, A74 immunoaffinity chromatography allowed to purify specific glycoproteins relevant to the SCO ventricular secretory process. The eluted fractions of the subcommissural organ (SCO), the cerebral hemispheres (CH) and the medulla oblongata (MO) were compared using the Concanavalin A (Con A) and wheat germ agglutinin (WGA) staining procedures after western-blotting. Analysis of the optical density of the reactive bands allowed to estimate the relative concentration of the various glycopeptides in the eluted fractions. In the SCO-eluted fractions at least ten Con A-positive glycopeptides were identified, their apparent molecular weight ranging from 240 to 42 kD. Only three of these appeared to be WGA-positive (98, 88, and 52 kD). In the CH-eluted fractions only a 52 kD Con A- and WGA-positive glycopeptide was revealed, while in the MO-eluted fractions a 32 kD glycopeptide was also Con A- and WGA-positive. These results are discussed in regard to the known biosynthesis pathway of complex type glycoproteins.

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“…In the ovine Meiniel et al (1986), detected after western blotting a ConA-positive 54kDa glycopeptide, specific of the SCO soluble extracts. Using electrophoresis, Sterba et al (1987) visualized more than 40 polypeptides ranging from 350 to 10 kDa in the bovine fiber of Reissner. Rodriguez et al (1987) used, in the same species, antibodies raised against Reissner's fiber (Antiserum Fiber of Reissner Urea = AFRU) to purify on immunoadsorbent components from SCO and RF extracts.…”

Section: Introductionmentioning

confidence: 99%

A comparative immunocytochemical and immunochemical analysis of glycoproteins synthesized in the bovine subcommissural organ

Karoumi

Meiniel

Belin

et al. 1991

J. Neural Transmission

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To extend our previous immunochemical investigations in the chick embryo (Karoumi et al., 1990 b), we raised antibodies in the rabbit against crude extracts of the subcommissural organ (SCO) of the bovine. The antiserum labeled A99 was absorbed by crude brain extracts and its specificity was tested by different techniques. Comparison of crude SCO and cerebral hemispheres supernatants after immunoblotting allow to identify specific 98, 60, 52, 42, 38, and 32 kDa polypeptides in the SCO profile. Immunoaffinity chromatography on A99 immunoadsorbent of crude SCO, cerebral hemispheres (CH) and classical ependyma (CE) supernatants was followed by electrophoretical analysis and electrotransfer. Concanavalin A (Con A) and wheat germ agglutinin (WGA) labeling procedures demonstrated the presence of numerous glycopeptides specific of crude SCO supernatants and having an apparent molecular weight ranging from 240 to 50 kDa. In the CH-eluted fraction, 50 and 52 kDa glycopeptides were revealed by ConA and WGA, whereas in the CE-immunopurified fraction no band was visualized. The similarity of the chick embryo and bovine electrophoretic pattern corresponding to the SCO eluted fractions speaks in favour of a high degree of conservation of the SCO secretory material and an evolutionary stability of the antigens recognized by A99IgG.

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The secretion of the subcommissural organ

Cited by 80 publications

References 4 publications

Immunochemical analysis of the subcommissural organ-Reissner's fiber complex using antibodies against alkylated and deglycosylated glycoproteins of the bovine Reissner's fiber

Immunochemical analysis of the subcommissural organ-Reissner's fiber complex using antibodies against alkylated and deglycosylated glycoproteins of the bovine Reissner's fiber

Glycoprotein synthesis in the subcommissural organ of the chick embryo II. an immunochemical study

A comparative immunocytochemical and immunochemical analysis of glycoproteins synthesized in the bovine subcommissural organ

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