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. 2020 May 26;10:8720. doi: 10.1038/s41598-020-64977-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Micronuclei in germ cells (gonocytes) of tadpoles contain individual chromosomes selectively eliminated in diploid (a,c,e,g) and triploid (b,d,f,h) hybrids. (a,b) Whole-mount immunofluorescent staining with CREST serum in tadpole gonads revealed one kinetochore in each micronucleus. (d,f) 3D-FISH with a probe specific to the P. ridibundus RrS1 repeat revealed P. ridibundus centromeres in the cell nuclei in the gonads of diploid and triploid hybrids as well as in the micronuclei in the gonads of triploid hybrids. (c,e) P. ridibundus chromosomes were usually absent from the micronuclei in the gonads of diploid hybrids. Images (a–f) are single confocal sections of 0.8 µm in thickness. (g,h) 3D surface reconstruction of the germ cells and micronuclei depicted in (e,f). (g) 3D surface reconstruction clearly demonstrates the absence of signals in the micronuclei and 13 P. ridibundus centromeres (red) in the interphase nuclei (gray) of diploid hybrids. (h) 3D surface reconstruction shows the presence of 1 P. ridibundus centromere (red) in each micronucleus and 9 P. ridibundus centromeres (red) in the interphase nucleus (gray) of triploid LLR hybrids. Arrowheads indicate micronuclei. Scale bars for images (a–d) = 20 µm. Scale bars for images (e–h) = 10 µm.