Fluorescence replication banding of frog chromosomes

Abstract

To identify individual chromosomes of a frog karyotype by their fluorescence banding patterns, chromosomes were stained with actinomycin D and 4,6-diamidino-2-phenylindole (DAPI) after incorporation of BrdU during the late S-phase. The chromosomes of three Rana species which were selected for this study (R. ridibunda, R. lessonae and R. japonica) showed well-defined late replication bands. The fluorescence patterns obtained were the reverse of those produced by a 4Na-EDTA Giemsa-staining technique. Fluorescence patterns of the two water frog species (R. ridibunda and R. lessonae) were similar to each other, except for the different fluorescence of the centromeric heterochromatin, which gave extremely bright signals in R. ridibunda but no signal in R. lessonae. Experiments also showed differences between the fluorescence patterns of R. lessonae chromosome 13 in the Italian and Luxembourgian populations. These results show that the fluorescence replication banding using actinomycin D and DAPI is very effective in identifying individual frog chromosomes and detecting their structural changes.

Features of the karyotypes of Pelophylax ridibundus Pallas, 1771 and Rana macrocnemis Boulenger, 1885 (Amphibia: Ranidae) from Armenia

“…Comparison of the chromosomal data of P. ridibundus with other species of the green frogs (Heppich, 1978;Schmid, 1978;Miura, 1995;Miura et al, 1997) and some species of brown frogs (Orlova et al, 1978;Birstein, 1984;Green, Borkin, 1993;Popov, Dimitrov, 1999) with data from this study confi rms the close similarity of several chromosome pairs by size and morphology. On the whole, the interspecifi c differences in karyotypes of species of Rana and Pelophylax are manifested in the distinctions of late replication banding patterns (Miura, 1995;Miura et al, 1997), fl uorescence replication banding patterns (Picariello et al, 2002) and in the locations of C-heterochromatin blocks (Schmid, 1978;Miura, 1995;Miura et al, 1997;present study).…”

“…On the whole, the interspecifi c differences in karyotypes of species of Rana and Pelophylax are manifested in the distinctions of late replication banding patterns (Miura, 1995;Miura et al, 1997), fl uorescence replication banding patterns (Picariello et al, 2002) and in the locations of C-heterochromatin blocks (Schmid, 1978;Miura, 1995;Miura et al, 1997;present study).…”

“…Taking into account the taxonomic status of these frogs and chromosome sets differentiation (m/st pairs), the differences between karyotypes of lake frogs in Turkey and Saudi Arabia can also (Heppich, 1978;Schmid, 1978;Belcheva et al, 1985;Miura, 1995;Miura et al, 1997), as the morphological elements of the karyotype vary both among populations of P. ridibundus as well as among the species of P. ridibundus complex. It is known that the several techniques features, such as C-banding patterns, fl uorescence replication banding patterns (DAPI and FISH) are used for an accurate distinction of the species of green and brown frogs (Schmid, 1978;Miura, 1995;Miura et al, 1997;Picariello et al, 2002). Hence, those should also be used for the fi nal confi rmation of differences in the frog species within the P. ridibundus complex.…”

Evidence for Integrity of Parental Genomes in the Diploid Hybridogenetic Water Frog <i>Pelophylax esculentus</i> by Genomic in situ Hybridization

“…Diploid karyotypes of P. lessonae , P. ridibundus , and P. esculentus are composed of 26 chromosomes, considered so far as morphologically similar in length and shape in all 3 taxa [Aleksandrovskaya, 1976;Heppich, 1978;Bucci et al, 1990;Miura et al, 1997]. Heppich et al [1982] firstly revealed that the genomes of diploid P. esculentus hybrids are composed of 2 groups of chromosomes with differential affinity to AMD-DAPI staining in pericentromeric heterochromatin.…”