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Cytophotometric determination of genome size in two species of Cyclops of Lake Baikal (Crustacea: Copepoda, Cyclopoida) in ontogenetic development
Authors:E. A. Ivankina  A. L. Alekseeva  V. F. Semeshin  L. V. Omelyanchuk  I. G. Palchikova  N. G. Sheveleva  S. V. Kirilchik  I. F. Zhimulev
Affiliation:16314. Institute of Molecular and Cellular Biology, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia
26314. Technological Design Institute of Scientific Instrument Engineering, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia
36314. Limnological Institute, Siberian Branch, Russian Academy of Sciences, Irkutsk, Russia
Abstract:The genome size of Cyclops in cells at early stages of cleavage (up to the fifth division) and in somatic cells was estimated by static digital Feulgen cytophotometry in order to study quantitative changes in DNA content during chromatin diminution. Described here cytophotometric method was approbated on five different digital-imaging systems in blood cells of four vertebrate species. In all cases, we observed a direct correlation between the data obtained with known from the literature on genome size and high reproducibility, which will allow these systems to be used in future work. We also optimized the conditions for DNA hydrolysis of both blood smears and for two species of Cyclops from the Moscow population as 30 min in 5 N HCl at 24°C. Here, we first revealed chromatin diminution in two endemic Baikal species of Cyclopoida: Acanthocyclops incolotaenia and Diacyclops galbinus. We estimated the extent of chromatin diminution in Diacyclops galbinus as 95.5–96.2%. Cytometric analysis of the third species, Mesocyclops leuckarti, did not reveal obvious chromatin diminution.
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