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Preparation of Xenopus tropicalis whole chromosome painting probes using laser microdissection and reconstruction of X. laevis tetraploid karyotype by Zoo-FISH

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Abstract

Laser microdissection was used for the preparation of whole chromosome painting probes in Silurana (Xenopus) tropicalis. Subsequent cross-species fluorescence in situ hybridization (Zoo-FISH) on its tetraploid relative Xenopus laevis revealed persistence of chromosomal quartets even after 50–65 million years of separate evolution. Their arrangement is in a partial concordance with previous experiments based on similarity of a high-resolution replication banding pattern. Further support for an allotetraploid origin of X. laevis was given by hybridization with a probe derived from the smallest X. tropicalis chromosome (Xt10). Here, pericentric areas of both arms of Xl 14 and 18 were stained, indicating intrachromosomal rearrangements. The positions of signals were not in agreement with the chromosomal quartets revealed by painting probes Xt 8 and 9 (Xl 11 + 14 and Xl 15 + 18, respectively). This suggests that both X. tropicalis chromosomes underwent non-reciprocal translocation of Xt10 separately in at least two different ancient ancestors. In addition, the observed translocation events could explain the origin of individuals with 18 chromosomes in diploid karyotypes, probably extinct after the genesis of the allotetraploid X. laevis (2n = 36).

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Abbreviations

DAPI:

4′,6-diamidino-2-phenylindole

DOP-PCR:

degenerate oligonucleotide primed polymerase chain reaction

FISH:

fluorescence in situ hybridization

MYA:

million years ago

NOR:

nucleolus organizer region

SSR:

simple sequence repeat

TSA:

tyramide signal amplification

WCP:

whole chromosome painting

WGA:

whole genome amplification

Xl:

Xenopus laevis chromosome

Xt:

Xenopus tropicalis chromosome

Zoo-FISH:

cross-species fluorescence in situ hybridization

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Acknowledgements

This work was supported by grant 304/07/P204 from the Grant Agency of the Czech Republic, by grant MSM0021620858 from the Ministry of Education, Youth and Sports of the Czech Republic and by project MZE 0002716202 of the Grant Agency of the Ministry of Agriculture of the Czech Republic.

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Authors and Affiliations

  1. Faculty of Science, Charles University in Prague, Vinicna 7, 128 44, Prague 2, Czech Republic

    Vladimir Krylov, Jaroslav Macha, Tereza Tlapakova, Eva Seifertova & Natasa Sebkova

  2. Veterinary Research Institute, Hudcova 70, 621 00, Brno, Czech Republic

    Svatava Kubickova & Jiri Rubes

Authors
  1. Vladimir Krylov
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  2. Svatava Kubickova
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  3. Jiri Rubes
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  4. Jaroslav Macha
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  5. Tereza Tlapakova
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  6. Eva Seifertova
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  7. Natasa Sebkova
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Correspondence to Vladimir Krylov.

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Responsible Editor: Walther Traut.

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Krylov, V., Kubickova, S., Rubes, J. et al. Preparation of Xenopus tropicalis whole chromosome painting probes using laser microdissection and reconstruction of X. laevis tetraploid karyotype by Zoo-FISH. Chromosome Res 18, 431–439 (2010). https://doi.org/10.1007/s10577-010-9127-x

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  • DOI: https://doi.org/10.1007/s10577-010-9127-x

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