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Immunohistochemical investigation of epithelial, mesenchymal, neuroectodermal, immune and endocrine markers in sterlet (Acipenser ruthenus), shortnose sturgeon (Acipenser brevirostrum) and common carp (Cyprinus carpio)

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Summary

Immunohistochemistry (IHC) is a laboratory method widely used to characterize tissue and cell origin, both in human and veterinary medicine. In fish, however, little is known about staining characteristics of most tissue types, and especially for less studied chondrostean fish. The aim of this study was to examine the specificity of various immunohistochemical markers in tissues of chondrostean and teleostean fish and to validate diagnostic tests. Sterlet (Acipenser ruthenus L.), shortnose sturgeon (Acipenser brevirostrum) and common carp (Cyprinus carpio L.) were examined. Markers were chosen as representatives of epithelial (cytokeratin AE1/AE3), mesenchymal (vimentin), neuroectodermal (S-100 protein), lymphoid (leukocyte common antigen, LCA) and endocrine (thyroglobulin, thyroxin) tissues and organs. Applied antibodies were of monoclonal or polyclonal mammalian origin and primarily intended for human medicine research or diagnostic application. No species differences were obvious while examining sterlet, shortnose sturgeon and carp. Cytokeratin AE1/AE3, vimentin, S-100 protein and thyroxin were positive on targeted tissues and structures. Leukocyte common antigen (LCA) and thyroglobulin were negative on targeted structures, however, and with clear cross-reactivity on non-targeted tissues (vascular wall, granulocytes). Conclusive results were obtained when using polyclonal antibodies with dilution adjusted to laboratory practice, while application of ready-to-use (RTU) kits with pre-diluted antibodies or monoclonal antibodies often showed conflicting or inconclusive results.

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Data availability

The manuscript has no associated data. Immunohistochemical slides are deposited in Eva Šálková and Christoph Steinbach repository.

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Acknowledgements

This output reflects the views only of the authors, and the European Union cannot be held responsible for any use that may be made of the information contained herein. The authors wish to acknowledge the technical support of Mrs. Marcela Staňková from AeskuLab Patologie, Prague and valuable advice of Mrs. Eliška Axmannová from the Department of Pathology and Molecular Medicine, 2nd Faculty of Medicine and Teaching Hospital in Motol, Charles University in Prague. We also wish to acknowledge the technical support and tissue histological processing by Mrs. Jitka Špačková (Tumová) from USB FFPW. The authors wish to acknowledge the language help of Gale A. Kirking from the Business and editorial office, English Editorial Services in Brno, Czech Republic.

Funding

The study was financially supported by the Ministry of Education, Youth and Sports of the Czech Republic projects ‘CENAKVA’ (LM2018099) and PROFISH (CZ.02.1.01/0.0/0.0/16_019/0000869).

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Authors and Affiliations

  1. Faculty of Fisheries and Protection of Waters, South Bohemian Research Centre of Aquaculture and Biodiversity of Hydrocenoses, University of South Bohemia in České Budějovice, Zátiší 728/II, 389 25, Vodňany, Czech Republic

    Eva Šálková, Hana Kocour Kroupová & Christoph Steinbach

  2. Centre for Fish and Wildlife Health, Department of Infectious Diseases and Pathobiology, University of Bern, Länggassstrasse 122, 3012, Bern, Switzerland

    Heike Schmidt-Posthaus

  3. Department of Ecophysiology and Aquaculture, Leibniz Institute of Freshwater Ecology and Inland Fisheries (IGB), Müggelseedamm 301, 12587, Berlin, Germany

    Ilka Lutz

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  1. Eva Šálková
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  4. Hana Kocour Kroupová
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Contributions

E.Š. provided the topic, goal and study management; histological data evaluation and interpretation, and supervision of the results obtained; further performed the sterlet sampling, manuscript preparation and manuscript revision. H.S. supervised the results obtained and participated in manuscript review. I.L. performed immunohistochemical examination and testing (especially thyroxin), established the guideline for thyroxin testing and participated in the manuscript revision. H.K.K. provided study management and supervision and participated in manuscript review. C.S. performed carp sampling, IHC examination and testing, histological data evaluation; and further provided study management and supervision, and participated in manuscript review.

Corresponding authors

Correspondence to Eva Šálková or Christoph Steinbach.

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Ethics approval and consent to participate

The fish originated from the hatchery of the Faculty of Fisheries and Protection of Waters (FFPW), University of South Bohemia (USB), Czech Republic. This study was carried out in compliance with Czech Law No. 246/1992, on Animal Welfare. All applied protocols were supervised by the Institutional Animal Care and Use Committee (IACUC) of the USB FFPW in Vodňany. The FFPW has approval from the Ministry of Agriculture of the Czech Republic for handling and usage of experimental animals (ref. no. 16OZ15759/2013–17214). In sacrificing the fish, all efforts were made to minimize suffering.

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Šálková, E., Schmidt-Posthaus, H., Lutz, I. et al. Immunohistochemical investigation of epithelial, mesenchymal, neuroectodermal, immune and endocrine markers in sterlet (Acipenser ruthenus), shortnose sturgeon (Acipenser brevirostrum) and common carp (Cyprinus carpio). Fish Physiol Biochem 48, 1737–1749 (2022). https://doi.org/10.1007/s10695-022-01145-6

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