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The large amphibian genomes contain numerous repetitive DNA components that have played an important role in the karyotypic diversification of this vertebrate group. Hypotheses based on the presumable primitive karyotype (2n = 20) of the anurans of the family Pipidae suggest that they have evolved principally through intrachromosomal rearrangements. Pipa is the only South American pipid, while all the other genera are found in Africa. The divergence of the South American lineages from the African ones occurred at least 136 million years ago and is thought to have had a strong biogeographic component. Here, we tested the potential of the repetitive DNA to enable a better understanding of the differentiation of the karyotype among the family Pipidae and to expand our capacity to interpret the chromosomal evolution in this frog family. Our results indicate a long history of conservation in the chromosome bearing the H3 histone locus, corroborating inferences on the chromosomal homologies between the species in pairs 6, 8, and 9. The chromosomal distribution of the microsatellite motifs also provides useful markers for comparative genomics at the chromosome level between Pipa carvalhoi and Xenopus tropicalis, contributing new insights into the evolution of the karyotypes of these species. We detected similar patterns in the distribution and abundance of the microsatellite arrangements, which reflect the shared organization in the terminal/subterminal region of the chromosomes between these two species. By contrast, the microsatellite probes detected a differential arrangement of the repetitive DNA among the chromosomes of the two species, allowing longitudinal differentiation of pairs that are identical in size and morphology, such as pairs 1, 2, 4, and 5. We also found evidence of the distinctive composition of the repetitive motifs of the centromeric region between the species analyzed in the present study, with a clear enrichment of the (CA) and (GA) microsatellite motifs in P. carvalhoi. Finally, microsatellite enrichment in the pericentromeric region of chromosome pairs 6, 8, and 9 in the P. carvalhoi karyotype, together with interstitial telomeric sequences (ITS), validate the hypothesis that pericentromeric inversions occurred during the chromosomal evolution of P. carvalhoi and reinforce the role of the repetitive DNA in the remodeling of the karyotype architecture of the Pipidae.
FIGURE 1. Metaphase chromosomes of Pipa carvalhoi submitted to fluorescent in situ hybridization with the histone H3 probe. The chromosome pairs with hybridization signals detected in both chromatids of each homolog are indicated by the arrowheads.
FIGURE 2. Metaphase chromosomes of Pipa carvalhoi
(A–C) and Xenopus tropicalis
(D–F) submitted to fluorescent in situ hybridization with probes for the microsatellite repeat motifs (GA)15
(A,D), (CA)15
(B,E), and (GATA)8
(C,F). The arrows indicate the minor hybridization signals detected in the non-terminal regions of the chromosomes.
FIGURE 3. Metaphase chromosomes of Pipa carvalhoi
(A,B) and Xenopus tropicalis
(C,D) submitted to fluorescent in situ hybridization with probes for the microsatellite repeat motifs (CAG)10
(A,C) and (CCG)10
(B,D). The arrows indicate the minor hybridization signals detected in the non-terminal regions of the chromosomes.
FIGURE 4. Metaphase chromosomes of Pipa carvalhoi submitted to fluorescent in situ hybridization with probes for the microsatellite repeats (GACA)4 (A) and (GAA)10 (B). The arrows indicate the minor hybridization signals detected in the non-terminal regions of the chromosomes.
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