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Production and characterisation of cell- and tissue-specific monoclonal antibodies for the flatworm Macrostomum sp.

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Abstract

Monoclonal antibodies (mABs) against various cell types of the basal free-living flatworm Macrostomum sp. were produced by immunising Balb/c mice with cell suspensions of disintegrated animals. We identified 360 positive supernatants with specific staining of various tissues, cell types, patterns or structures. Here we report immunocytochemical characterisation, histological stainings and isotyping of 11 mABs specific for muscle cells (MMu-1, MMu-2, MMu-3, MMu-4), digestive and prostate glands (MDr-1 and MDr-2, MPr-1), epidermal cells (MEp-1), the ventral nerve cord including neuron clusters (MNv-1), gastrodermal cells (MDa-1) and spermatids (MSp-1). Confocal microscopy, histological techniques, electron microscopy and immunoblotting were applied to demonstrate stainings in juveniles, adults, starved or well-fed animals. Considering the current lack of specific markers the obtained mABs will be particularly helpful studying embryonic and postembryonic development, pattern formation, cell differentiation, regeneration and reproductive allocation in Macrostomum sp., and possibly other basal flatworms. The small size, ease of culturing, short generation time, transparency and the basal phylogenetic position specify Macrostomum sp. as a suitable model organism for comparative analyses within Platyhelminthes and to Drosophila and C. elegans.

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Acknowledgements

We are grateful to Peter Berger for the support and suggestions on the hybridoma technique and Bernd Pelster for the access to the confocal microscope. We thank Dr. Hermann Dietrich for support with immunisations and Renate Weiler-Görz and Dr. Alexandra Lusser for help with the immunoblot assays. This work was supported by FWF grants P13060 and P15204. L.S is supported by a Liese-Meitner fellowship (FWF, Austria). P.L. is supported by an APART fellowship (number 10841) of the Austrian Academy of Sciences.

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Authors and Affiliations

  1. Institute of Zoology and Limnology, University of Innsbruck, Technikerstrasse 25, 6020, Innsbruck, Austria

    Peter Ladurner, Daniela Pfister, Christof Seifarth, Lukas Schärer, Monika Mahlknecht, Willi Salvenmoser & Reinhard Rieger

  2. Institute for Biomedical Aging Research, Austrian Academy of Sciences, Rennweg 10, 6020, Innsbruck, Austria

    Regine Gerth

  3. Department of Molecular Biology, Innsbruck Medical University, Peter-Mayr Strasse 4b/III, 6020, Innsbruck, Austria

    Florentine Marx

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  1. Peter Ladurner
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  2. Daniela Pfister
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  3. Christof Seifarth
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Correspondence to Peter Ladurner.

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Ladurner, P., Pfister, D., Seifarth, C. et al. Production and characterisation of cell- and tissue-specific monoclonal antibodies for the flatworm Macrostomum sp.. Histochem Cell Biol 123, 89–104 (2005). https://doi.org/10.1007/s00418-004-0722-9

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