Determination of cellular DNA content of Iberian salamanders by flow cytometry

Abstract

The DNA content per diploid nucleus (2 C-value) was determined for the eight species of Iberian salamanders by flow cytometry. All species showed high values, which are characteristic of Caudata. The species with the lowest value was Pleurodeles waltl (48.25 pg DNA/cell) and Salamandra salamandra had the highest values (70.55 pg DNA/cell). There were no statistical differences between sexes in any of the eight species and DNA content was not directly related with phylogeny. The values obtained are often higher than previously published estimates obtained with older and potentially less accurate methodologies. We discuss the possible relationship between cellular DNA content and reproductive biology. Se determinó el contenido en ADN por núcleo diploide (valor 2-C) por citometría de flujo de ocho especies de salamandras ibéricas. Todas las especies mostraron altos valores, lo que es característico de Caudata. La especie con el menor valor fue Pleurodeles waltl (48,25 pg ADN/célula) mientras Salamandra salamandra tuvo las valores más altos (70,55 pg ADN/célula). No se encontraron diferencias estadísticas entre sexos en ninguna de las 8 especies y el contenido en ADN no se relacionó directamente con la filogenia. Los valores obtenidos son, a menudo, más altos que los publicados previamente con técnicas más antigüas, y potencialmente menos precisas. Discutimos las posibles relaciones entre el contenido en ADN celular y la biología reproductiva de cada especie.

Reading and editing the Pleurodeles waltl genome reveals novel features of tetrapod regeneration

“…Our estimates agree with Licht and Lowcock (2 C value = 42.7 pg) 53 , Morescalchi and Olmo (2 C value = 39 pg) 54 and are lower than Lizana and collegues (2 C value = 48.25 pg) 55 .…”

Flow cytometry measurement of the DNA contents of G0/G1 diploid cells from three different teleost fish species

“…The retinal tissue was placed in a sodium citrate buffer (250 nM sucrose (Sigma, St. Louis, MO), trisodium citrate 40 mM (Sigma), and dimethylsulfoxide 5% (v/v; Merck, Frankfurter, Darmstadt, Germany), pH adjusted to 7.6). Single‐cell suspensions were obtained immediately from the retinas using mechanical disaggregation procedures according to well‐established protocols (13–16). The sample was then centrifuged, the supernatant discarded, and the cell pellet resuspended in the sodium citrate buffer.…”

“…The DNA cell content of each retinal cell analyzed was calculated by dividing the modal fluorescence intensity obtained for the G0/G1 peak of retinal cells by the modal fluorescence intensity of standard human leukocytes (16). To translate relative DNA cell content into picograms of DNA per diploid cell nucleus (2n value), standard human leukocytes were considered to have 7.0 pg of DNA per nucleus (9, 17).…”

Taxonomic status of Triturus vittatus (Amphibia: Salamandridae) in western Turkey: trunk vertebrae count, genome size and allozyme data

“…Among amphibians, the highest level of within-population variation was recorded for Plethodon vehiculum (CV = 7.5%, data of Licht and Lowcock, 1991 were re-calculated by us). Usually, however, individual variation in amphibian species was considerably lower (Licht and Lowcock, 1991;MacCulloch et al, 1996;Murphy et al, 1997;Lizana et al, 2000). For instance, in populations of salamandrids CVs ranged from 0.1% to 1.7% (mean is 0.64 ± 0.03), and were very similar to that of hynobiids, pelobatids, and some other amphibians studied in our laboratory at the same conditions (Litvinchuk et al, 1997(Litvinchuk et al, , 1999(Litvinchuk et al, , 2004Rosanov and Vinogradov, 1998;Borkin et al, 2001; our data).…”