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  • 1
    In: Alternatives to Laboratory Animals, SAGE Publications, Vol. 28, No. 4 ( 2000-07), p. 575-601
    Abstract: Neuropeptides such as cholecystokinin (CCK) are subjected to a stepwise enzymic degradation (peptide processing) during axonal transport from the somata to the nerve terminals. This results in a continual change in the primary, secondary and tertiary structures of the peptide. Thus, an antibody raised against a selected sequence of the propeptide may recognise the antigen only at a certain stage of its “ontogenesis”. To address these difficulties, a set of antibodies with differing specificities and origins (chicken, rabbit) were used to visualise neuronal CCK by immunohistochemical methods in rat–brain sections (RBS) and in rat primary neuronal cultures (PNC). The specificity of the antibodies was analysed by using a dot-blot assay and a radioimmunoassay. Marked differences in the reactivities of the various antibodies were observed and related to the antigen used, inter-individual variations and the animal species. In the cortex, various types of CCK-immunoreactive neurons were found, including spindle-shaped or button-shaped neurons and pseudo-unipolar neurons. However, in contrast to mammalian antibodies, several of the chicken antibodies recognised cortical pyramidal neurons in both RBS and PNC without pretreatment with colchicine. Evidence has been obtained in both RBS and PNC that an antibody with a defined specificity may not visualise the entire neuron, but only distinct parts of it, possibly depending on the actual molecular structure of the neuropeptide present at a specific locus of the neuron. A complete mapping of a neuronal peptide that is processed during axonal transport can only be achieved by using a set of different antibody-specificities.
    Type of Medium: Online Resource
    ISSN: 0261-1929 , 2632-3559
    Language: English
    Publisher: SAGE Publications
    Publication Date: 2000
    detail.hit.zdb_id: 2390905-5
    SSG: 12,22
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  • 2
    In: Alternatives to Laboratory Animals, SAGE Publications, Vol. 25, No. 5 ( 1997-09), p. 555-586
    Abstract: It has been known for over a century that specific antibodies can be extracted from the eggs of immunised chickens. However, it was only when animal welfare became a subject of public debate that the chicken was considered as an alternative source of antibodies due to the possibility of non-invasive antibody sampling. Unfortunately, the welfare of animals alone is not sufficient to attract the interest of scientists; it is therefore important to demonstrate to potential users that avian antibodies can be used successfully in a variety of scientific investigations. The particular specificity of avian antibodies would appear to be due to the phylogenetic difference between Mammalia and Aves as well as to differences between the molecular structures of avian immunoglobulin (IgY) and mammalian immunoglobulin (IgG). The use of avian antibodies has additional advantages, as a considerable quantity of antibodies can be obtained from one chicken, and because the specificity of avian antibodies often markedly differs from that of comparable mammalian antibodies. This paper aims to demonstrate the advantages of using avian antibodies by presenting the results of five separate studies. In the first study, coordinated by Rüdiger Schade, the visualisation of cholecystokinin-like immuno-reactivity in the substantia nigra of rats by using anti-cholecystokinin antibody, without the pre-treatment of colchicine, is described. The second study, headed by Albrecht Zott, describes the use of avian antibodies in the identification of modern acellular pertussis vaccines by using rocket immunoelectrophoresis. The identification of unknown vaccine batches and the comparison with reference vaccines is a prerequisite for reducing the number of animal experiments necessary for vaccine control. The third study, coordinated by Martin Gerl, investigates the specificity of antibodies directed against the N-terminal propeptide of procollagen type III (PIIINP). Among the antibodies originating from different species (rabbit, mouse and chicken), only the chicken antibody was able to respond to the PIIINP in both human and rat sera. Thus, a direct comparison between human serum samples (alcoholic liver) and serum samples derived from corresponding animal models was possible. The fourth study, coordinated by Michael Erhard, shows that egg yolk antibodies can be successfully used to manage infectious diarrhoea in young agricultural animals. The final study, led by Andreas Hlinak, describes the successful production of anti-bovine leukaemia virus antibody. This antibody could be used in several diagnostic systems (for example, enzyme immunoassays and cytology). The five studies demonstrate that avian antibodies are an attractive alternative to mammalian antibodies, not only with respect to the welfare of animals, but also with respect to scientific and economic considerations.
    Type of Medium: Online Resource
    ISSN: 0261-1929 , 2632-3559
    Language: English
    Publisher: SAGE Publications
    Publication Date: 1997
    detail.hit.zdb_id: 2390905-5
    SSG: 12,22
    Location Call Number Limitation Availability
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  • 3
    Online Resource
    Online Resource
    SAGE Publications ; 2000
    In:  Alternatives to Laboratory Animals Vol. 28, No. 6 ( 2000-11), p. 815-831
    In: Alternatives to Laboratory Animals, SAGE Publications, Vol. 28, No. 6 ( 2000-11), p. 815-831
    Abstract: The in vitro culture of various cell types is an important scientific tool and is becoming increasingly acceptable as a viable alternative to animal experiments. Fetal calf serum (FCS) is a supplement used in many cell culture media, and provides cells with growth factors and cytokines necessary for successful culture. In view of the animal welfare issues surrounding the production of FCS, an alternative agent allowing the replacement or reduction in the use of FCS is desirable. A yolk extract factor, egg yolk factor X (EYF-X), obtained from chicken eggs is described, which facilitates the in vitro culture of a variety of cell types. When the extract was added to a culture medium used for in vitro fertilisation, the number of successful fertilisations was significantly increased. In a further in vitro model (permanent neuronal cell line N2A), the yolk extract significantly stimulated cell proliferation, as well as the growth of cell processes. A set of specific antibodies against different parts of the prepro-cholecystokinin reacted with the extract. The intensity of the reaction depended on the age of the egg (time after the laying date). Analysis by gel chromatography recorded a main protein fraction with an apparent molecular mass of 20–30kDa. This fraction was labelled by Western blot with an antibody with specificity against CCK-octapeptide. These findings suggest that the yolk factor may be a CCK/gastrin-like molecule. Since CCK/gastrin-like molecules have also been detected in the spermatozoa of mammals, the influence on in vitro fertilisation could be explained by the yolk factor replacing the endogenous CCK/gastrin-like molecule destroyed in sperm freezing. The results of this study suggest that it might be possible to replace FCS with EYF-X. The application of the yolk factor to a broad spectrum of cell types remains to be investigated.
    Type of Medium: Online Resource
    ISSN: 0261-1929 , 2632-3559
    Language: English
    Publisher: SAGE Publications
    Publication Date: 2000
    detail.hit.zdb_id: 2390905-5
    SSG: 12,22
    Location Call Number Limitation Availability
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  • 4
    Online Resource
    Online Resource
    SAGE Publications ; 1991
    In:  Alternatives to Laboratory Animals Vol. 19, No. 4 ( 1991-10), p. 403-419
    In: Alternatives to Laboratory Animals, SAGE Publications, Vol. 19, No. 4 ( 1991-10), p. 403-419
    Abstract: Based on a physiological process, antibodies are concentrated in the yolk of chicken eggs, and can be used as a convenient source of polyclonal antibodies. Hens were immunised with a BSA-cholecystokinin octapeptide conjugate (CCK-8) to obtain antibodies against CCK-8. A simple method is described for extracting the antibodies from egg yolk, which can then successfully be used in immunocytochemical studies. Using these antibodies, CCK-like immunoreactivity was found in brain sections of rats, both by fluorescein isothiocyanate and peroxidase staining. The specificities of these antibodies were different from those of rabbit antibodies against the C-terminal part of CCK-8. The applicability of egg yolk antibodies in immunological investigations is reviewed and discussed in detail. The properties of avian egg yolk antibodies are compared with those of polyclonal antibodies obtained from mammals (e.g. the rabbit). The more general use of avian antibodies is emphasised as a real alternative, taking into consideration the reduction in animal suffering involved in comparison with the classical immunisation techniques used on rabbits.
    Type of Medium: Online Resource
    ISSN: 0261-1929 , 2632-3559
    Language: English
    Publisher: SAGE Publications
    Publication Date: 1991
    detail.hit.zdb_id: 2390905-5
    SSG: 12,22
    Location Call Number Limitation Availability
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