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Continuous delivery of a monoclonal antibody against Reissner’s fiber into CSF reveals CSF-soluble material immunorelated to the subcommissural organ in early chick embryos

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Abstract

The subcommissural organ (SCO) is an ependymal differentiation located in the dorsal midline of the caudal diencephalon under the posterior commissure. SCO cells synthesize and release glycoproteins into the cerebrospinal fluid (CSF) forming a threadlike structure known as Reissner’s fiber (RF), which runs caudally along the ventricular cavities and the central canal of the spinal cord. Numerous monoclonal antibodies have been raised against bovine RF and the secretory material of the SCO. For this study, we selected the 4F7 monoclonal antibody based on its cross-reactivity with chick embryo SCO glycoproteins in vivo. E4 chick embryos were injected with 4F7 hybridoma cells or with the purified monoclonal antibody into the ventricular cavity of the optic tectum. The hybridoma cells survived, synthesized and released antibody into the CSF for at least 13 days after the injection. E5 embryos injected with 4F7 antibody displayed precipitates in the CSF comprising both the monoclonal antibody and anti-RF-positive material. Such aggregates were never observed in control embryos injected with other monoclonal antibodies used as controls. Western blot analysis of CSF from E4-E6 embryos revealed several immunoreactive bands to anti-RF (AFRU) antibody. We also found AFRU-positive material bound to the apical surface of the choroid plexus primordia in E5 embryos. These and other ultrastructural evidence suggest the existence of soluble SCO-related molecules in the CSF of early chick embryos.

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Acknowledgements

The authors are grateful to Dr. de la Rosa (Centro de Investigaciones Biológicas, CSIC, Madrid, Spain) for providing rabbit polyclonal antibody against Ng-CAM, José Esteban Casares Mira and Pedro Jiménez Palomo for valuable technical assistance, David Navas Fernández for technical assistance in the confocal microscopy study and Gregorio Martín Caballero for technical assistance with transmission electron microscopy.

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  1. E. Miranda

    Present address: Cambridge Institute for Medical Research, Department of Medicine, University of Cambridge, Room 6.36 Wellcome Trust/MRC Building, Hills Road, CB2 2XY, Cambridge, UK

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  1. Departamento de Biología Celular, Genética y Fisiología, Facultad de Ciencias, Campus de Teatinos, Universidad de Málaga, 29071, Málaga, Spain

    C. Hoyo-Becerra, M. D. López-Ávalos, J. Pérez, E. Miranda, P. Rojas-Ríos, P. Fernández-Llebrez & J. M. Grondona

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  1. C. Hoyo-Becerra
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Correspondence to J. M. Grondona.

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C. Hoyo-Becerra and M.D. López-Ávalos contributed equally to this study and should be considered as first authors.

C. Hoyo-Becerra was the recipient of a predoctoral fellowship (PFPI) from the Ministerio de Educacion y Cultura (Spain). This work was supported by grants from DGICYT (BFI2003-03348; Spain) and FIS (01/0948; Spain), FIS (01-0948, PI021517; Spain) and ISCIII (red CIEN, nodo Fundación Carlos Haya).

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Hoyo-Becerra, C., López-Ávalos, M.D., Pérez, J. et al. Continuous delivery of a monoclonal antibody against Reissner’s fiber into CSF reveals CSF-soluble material immunorelated to the subcommissural organ in early chick embryos. Cell Tissue Res 326, 771–786 (2006). https://doi.org/10.1007/s00441-006-0231-3

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