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Demonstration of primary cilia and acetylated α-tubulin in fish endothelial, epithelial and fibroblast cell lines

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Abstract

Primary cilia (PC) were demonstrated for the first time in fish endothelial, epithelial and fibroblast cell lines through immunofluorescence staining with the monoclonal antibody, 6-11B-1, against acetylated α-tubulin. The study was carried out with eight recently developed cell lines from the walleye, Sander vitreus (Mitchill). These were three fibroblast-like cell lines, WE-cfin11f, WE-skin11f and WE-liver3 from, respectively, the caudal fin, skin and liver, and three epithelial-like cell lines, WE-cfin11e, WE-spleen6 and WErpe from, respectively, the caudal fin, spleen and retina. Also, endothelial-like WEBA from the bulbus arteriosus and glial-like WE-brain5 from the brain were used. Immunocytochemistry revealed strong staining for acetylated α-tubulin in mitotic spindles and midbodies for all cell lines, and in PC for all cell lines except WE-skin11f. Staining of cytoplasmic microtubules (fibrils) was absent in three cell lines, including WEBA, but present in the others, especially WE-skin11f, which might have obscured PC detection in these cells. Tubacin, an inhibitor of histone deacetylase 6, induced cytoplasmic fibrils in WEBA and the intensity of their staining in WE-cfin11f. These results suggest that the cell lines might differ in their deacetylase activities, making them useful for studying this tubulin modification in teleosts, as well as for studying PC.

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Acknowledgments

This research was supported by a Discovery grant from the Natural Sciences and Engineering Research Council (NSERC) of Canada to NCB.

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  1. Department of Biology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada

    Nguyen T. K. Vo & Niels C. Bols

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Correspondence to Nguyen T. K. Vo.

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Vo, N.T.K., Bols, N.C. Demonstration of primary cilia and acetylated α-tubulin in fish endothelial, epithelial and fibroblast cell lines. Fish Physiol Biochem 42, 29–38 (2016). https://doi.org/10.1007/s10695-015-0114-1

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