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, 8 (3), 233-48
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Molecular Cytogenetic Analysis of the Crucian Carp, Carassius Carassius (Linnaeus, 1758) (Teleostei, Cyprinidae), Using Chromosome Staining and Fluorescence in Situ Hybridisation With rDNA Probes

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Molecular Cytogenetic Analysis of the Crucian Carp, Carassius Carassius (Linnaeus, 1758) (Teleostei, Cyprinidae), Using Chromosome Staining and Fluorescence in Situ Hybridisation With rDNA Probes

Aneta Spoz et al. Comp Cytogenet.

Abstract

The crucian carp Carassius carassius (Linnaeus, 1758) is a species with restricted and decreasing distribution in Europe. Six males and six females of the species from the Baltic Sea basin in Poland were examined to show sequentially CMA3/AgNO3 staining pattern, DAPI staining, and, for the first time in literature, molecular cytogenetic analysis using double-colour fluorescence in situ hybridisation (FISH) with 28S and 5S rDNA probes. The karyotype consisted of 20 m, 36 sm and 44 sta chromosomes, NF=156. The AgNO3 stained NORs were most frequently located terminally in the short arms of two sm and two sta elements, and CMA3-positive sites were also observed suggesting abundant GC-rich repetitive DNA in the regions. Other CMA3-positive sites in the short arms of six to ten sm and sta chromosomes were detected. The results based on 28S rDNA FISH confirmed the location of rDNA sites. DAPI-negative staining of NORs suggested the scarcity of AT-rich DNA in the regions. FISH with 5S rDNA probe revealed 8-14 loci (ten and 12 in respectively 49 and 29% of metaphases). They were located in two sm and eight to ten sta chromosomes and six of them were larger than others. Simultaneously, mapping of the two rDNA families on the chromosomes of C. carassius revealed that both 28S and 5S rDNA probes were located in different chromosomes. Molecular cytogenetic data of C. carassius presented here for the first time give an important insight into the structure of chromosomes of this polyploid and declining species and may be useful in its systematics.

Keywords: CMA3; Cyprinidae; FISH with rDNA; NOR-phenotype; molecular cytogenetics; polyploid species.

Figures

Figure 1.
Figure 1.
Giemsa stained metaphase (a), corresponding karyotype of Carassius carassius (b), and metaphase spread sequentially stained with AgNO3 (c) and CMA3 (d). NOR chromosomes shown in frames (in a and b), Ag-NORs and corresponding CMA3-positive sites shown by thick arrows (in c and d) and shown in inset (in d), other CMA3–positive sites shown by thin arrows (in d).
Figure 2.
Figure 2.
Metaphase fig of Carassius carassius DAPI stained (a) and with a single colour FISH (b) with 28S rDNA probe. 28S rDNA hybridisation signals shown by arrows.
Figure 3.
Figure 3.
Representative mitotic metaphase figs (a−d) and corresponding karyotype of Carassius carassius (e): a DAPI stained and b−d most frequent hybridisation pattern after dual colour FISH with four 28S rDNA sites (b), ten 5S rDNA sites (c) and both rDNA probes (d). Six stronger and four weaker 5S rDNA hybridisation sites (c) shown by thick and thin arrows, respectively.

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