Delimiting the Origin of a B Chromosome by FISH Mapping, Chromosome Painting and DNA Sequence Analysis in Astyanax paranae (Teleostei, Characiformes)

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2014-04-15

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Silva, Duilio M. Z. de A. [UNESP]
Pansonato-Alves, Jose Carlos [UNESP]
Utsunomia, Ricardo [UNESP]
Araya-Jaime, Cristian [UNESP]
Ruiz-Ruano, Francisco J.
Daniel, Sandro Natal [UNESP]
Hashimoto, Diogo Teruo [UNESP]
Oliveira, Claudio [UNESP]
Camacho, Juan Pedro M.
Porto-Foresti, Fabio [UNESP]

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Public Library Science

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Supernumerary (B) chromosomes have been shown to contain a wide variety of repetitive sequences. For this reason, fluorescent in situ hybridisation (FISH) is a useful tool for ascertaining the origin of these genomic elements, especially when combined with painting from microdissected B chromosomes. In order to investigate the origin of B chromosomes in the fish species Astyanax paranae, these two approaches were used along with PCR amplification of specific DNA sequences obtained from the B chromosomes and its comparison with those residing in the A chromosomes. Remarkably, chromosome painting with the one-arm metacentric B chromosome probe showed hybridization signals on entire B chromosome, while FISH mapping revealed the presence of H1 histone and 18S rDNA genes symmetrically placed in both arms of the B chromosome. These results support the hypothesis that the B chromosome of A. paranae is an isochromosome. Additionally, the chromosome pairs Nos. 2 or 23 are considered the possible B chromosome ancestors since both contain syntenic H1 and 18S rRNA sequences. The analysis of DNA sequence fragments of the histone and rRNA genes obtained from the microdissected B chromosomes showed high similarity with those obtained from 0B individuals, which supports the intraspecific origin of B chromosomes in A. paranae. Finally, the population hereby analysed showed a female-biased B chromosome presence suggesting that B chromosomes in this species could influence sex determinism.

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Plos One. San Francisco: Public Library Science, v. 9, n. 4, 10 p., 2014.