Antibody-mediated lysis of the bovine subcommissural organ maintained in culture

Cifuentes, M.; Rodríguez, E. M.; Hernández, S.; Pérez, Juan; Peruzzo, Bruno; Pérez-Fígares, J. M.; Fernández-Llébrez, Pedro

doi:10.1007/bf00228015

Cited by 3 publications

(3 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A remarkable feature of our grafted SCO was the process of circularization. Bovine SCO explants from adult animals Lehmann and Sterba 1993;Cifuentes et al 1995;Schoebitz et al 2001) acquire a round configuration, but with the apical membrane of ependymal cells towards the external surface of the explant. However, our embryonic explants adopted an inwards circular conformation.…”

Section: Secretory Activity Of the Grafted Scosmentioning

confidence: 99%

Reissner’s fiber formation depends on developmentally regulated factors extrinsic to the subcommissural organ

et al. 2005

View full text Add to dashboard Cite

Reissner's fiber (RF) is a threadlike structure present in the third and fourth ventricles and in the central canal of the spinal cord. RF develops by the assembly of glycoproteins released into the cerebrospinal fluid (CSF) by the subcommissural organ (SCO). SCO cells differentiate early during embryonic development. In chick embryos, the release into the CSF starts at embryonic day 7 (E7). However, RF does not form until E11, suggesting that a factor other than release is required for RF formation. The aim of the present investigation was to establish whether the factor(s) triggering RF formation is (are) intrinsic or extrinsic to the SCO itself. For this purpose, SCO explants from E13 chick embryos (a stage at which RF has formed) were grafted at two different developmental stages. After grafting, host embryos were allowed to survive for 6-7 days, reaching E 9 (group 1) and E13 (group 2). In experimental group 1, the secretion released by the grafted SCOs never formed a RF; instead, it aggregated as a flocculent material. In experimental group 2, grafted SCO explants were able to develop an RF-like structure, similar to a control RF. These results suggest that the factor triggering RF formation is not present in the SCO itself, since E13 SCO secretion forms an RF in E13 brains but never develops RF-like structures when placed in earlier developmental environments. Furthermore, the glycoproteins released by implanted SCOs bind specifically to several structures: the apical portion of the mesencephalic floor plate and the choroid plexus of the third and fourth ventricles.

show abstract

Section: Secretory Activity Of the Grafted Scosmentioning

confidence: 99%

Reissner’s fiber formation depends on developmentally regulated factors extrinsic to the subcommissural organ

et al. 2005

View full text Add to dashboard Cite

show abstract

“…An extract of bovine nervous tissue containing non-secretory ependyma from the lateral ventricle, plus corpus callosum, was prepared in the same way to be used as a negative control. (2) Conditioned medium of bovine SCO explants prepared according to Cifuentes et al (1995). The negative control was culture medium conditioned by explants of bovine non-secretory ependyma from the lateral ventricle.…”

Section: Antigen-mediated Elisamentioning

confidence: 99%

Analysis and quantification of the secretory products of the subcommissural organ by use of monoclonal antibodies

Fernández-Llébrez

Miranda

Estivill‐Torrús

et al. 2001

Microsc. Res. Tech.

View full text Add to dashboard Cite

Bovine ReissnerЈs fiber (RF) glycoproteins were used as antigen for the production of polyclonal and monoclonal antibodies (Mabs). We also produced Mabs against intracellular secretory glycoproteins of the bovine subcommissural organ (SCO). These Mabs were used for immunodetection of secretory proteins in situ (structural and ultrastructural immunocytochemistry), in blots, and in solutions. Three different antigen-mediated ELISA were designed to evaluate the affinity of the Mabs, to study the nature of the epitopes, and for competition test among Mabs. Two double antibody sandwich ELISA were designed to detect and quantify soluble secretory materials in different samples, to study coexistence of epitopes, and to elucidate whether epitopes for Mabs are repeated or not in the RF-glycoproteins. Twenty-three Mabs recognizing the bovine RF-and SCO-glycoproteins in solutions (ELISA) as well as in tissue sections, were obtained. Nineteen of these Mabs also recognized the pig SCO, 11 the rabbit SCO, 6 the dog SCO, and 5 the rat SCO. None of the Mabs recognized the SCO of non-mammalian species. The different types of ELISA demonstrated that: (1) the epitopes reside in the proteinaceous moiety of the secretion, (2) they coexist in the same molecular forms and, with few exceptions, they did not overlap, (3) they were not repeated in the secretory molecule(s). Three Mabs were used for immunoblotting of RF; one of them revealed the same band pattern as that shown by an anti-RF serum. It is concluded that all Mabs raised in our laboratory are directed against non-repeated sequences of RF-glycoproteins that have not been conserved in vertebrate phylogeny.

show abstract

“…Culture medium supernatant of bovine SCO explants. Explants from adult bovine SCO were prepared according to the procedure described by Cifuentes et al (1995) with minor modifications. Ten explants per SCO were obtained.…”

Section: Test Samplesmentioning

confidence: 99%

Quantification of the secretory glycoproteins of the subcommissural organ by a sensitive sandwich ELISA with a polyclonal antibody and a set of monoclonal antibodies against the bovine Reissner's fiber

Estivill‐Torrús

Cifuentes

Grondona

et al. 1998

Cell and Tissue Research

Self Cite

View full text Add to dashboard Cite

The subcommissural organ (SCO) is an ependymal brain gland that releases glycoproteins into the ventricular cerebrospinal fluid where they condense to form the Reissner's fiber (RF). We have developed a highly sensitive and specific two-antibody sandwich enzyme-linked immunosorbent assay (ELISA) for the quantification of the bovine SCO secretory material. The assay was based on the use of the IgG fraction of a polyclonal antiserum against the bovine RF as capture antibody and a pool of three peroxidase-labeled monoclonal antibodies that recognize non-overlapping epitopes of the RF glycoproteins as detection antibody. The detection limit was 1 ng/ml and the working range extended from 1 to 4000 ng/ml. The calibration curve, generated with RF glycoproteins, showed two linear segments: one of low sensitivity, ranging from 1 to 125 ng/ml, and the other of high sensitivity between 125 and 4000 ng/ml. This assay was highly reproducible (mean intra- and interassay coefficient of variation 2.2% and 5.3%, respectively) and its detectability and sensitivity were higher than those of ELISAs using exclusively either polyclonal or monoclonal antibodies against RF glycoproteins. The assay succeeded in detecting and measuring secretory material in crude extracts of bovine SCO, culture medium supernatant of SCO explants and incubation medium of bovine RF; however, soluble secretory material was not detected in bovine cerebrospinal fluid.

show abstract

Antibody-mediated lysis of the bovine subcommissural organ maintained in culture

Cited by 3 publications

References 27 publications

Reissner’s fiber formation depends on developmentally regulated factors extrinsic to the subcommissural organ

Reissner’s fiber formation depends on developmentally regulated factors extrinsic to the subcommissural organ

Analysis and quantification of the secretory products of the subcommissural organ by use of monoclonal antibodies

Quantification of the secretory glycoproteins of the subcommissural organ by a sensitive sandwich ELISA with a polyclonal antibody and a set of monoclonal antibodies against the bovine Reissner's fiber

Contact Info

Product

Resources

About