[Age Dynamics of Telomere Length in Endemic Baikal Planarians].

Age-related changes in telomere length (TL) in somatic tissues are not limited only to shortening. It is known that many organisms show different TL dynamics. Such species specificity indicates the complexity of the mechanisms involved in the regulation of TL. Owing to their morphological, physiological, and ecological features, Baikal planarians are an interesting model for studying the TL dynamics and the factors influencing it in comparison with species living outside Baikal. In this work, we investigated telomerase activity and age-related changes in TL in three endemic species of planarians from the Dendrocoelidae family. Two species are giant deep-water species (7-12 cm long, Sorocelis hepatizon and Rimacephalus arecepta), and one is a coastal shallow species (1 cm long, Baikalobia guttata). In addition, we investigated the telomere biology in another small Siberian species from the Planariidae family (2 cm in length, Phagocata sibirica), which is not found in Baikal. TL and telomerase activity were determined using real-time PCR and the TRAP method. Three types of age-related TL dynamics were detected with active telomerase: (1) TL shortening at the juvenile stage of development and subsequent maintenance (R. arecepta, Ph. sibirica), (2) gradual TL shortening during ontogeny (S. hepatizon) and (3) cyclic dynamics of TL (B. guttata). Thus, the changes of TL in the studied planarians does not have an obvious connection with body size, habitat depth, phylogenetic relationship and is probably a consequence of species features in the regulation of telomerase activity.

[Telomere length and phylogenetic relationship of Baikal and Siberian planarians (Turbellaria, Tricladida)].

Dynamics of the telomeric DNA (tDNA) and the phylogeny of the Baikal and Siberian planarians have been studied based on the analysis of the 18S rDNA and beta-actin gene fragments. A relationship between tDNA and the planarians size has been demonstrated. Giant planarians with a minor exception have longer tDNA than little planarians. Phylogenetic affinity between the species that have the stretched tracks of tDNA, big size and similar habitats may indicate possible role of tDNA in the development of the indefinite regenerative capacity of planarians.

[Interspecific variability of telomeric DNA length in some Siberian and endemic Baikal planarians (Plathelminthes, Tricladida)].

The length of the telomeric DNA in nine species of planarians inhabiting Lake Baikal and one Siberian species from Baikal rivers was determined using Southern hybridization. According to preliminary estimations, it varied in the range of 25-30 kb (Rimacephalus arecepta, Rimacephalus pulvinar, Sorocelis hepatizon, Sorocelis nigrofasciata, Protocotylus sp., Baikalobia guttata, Bdellocephala baikalensis, Phagocata sibirica) and 50 kb (Baikaloplana valida, Baikalobia copulatrix). It is the first estimation of the values of telomeric region lengths for Baikal free-living flat worms.

[Molecular-phylogenetic analysis of cyclopoids (Copepoda: Cyclopoida) from Lake Baikal and its water catchment basin].

Baikalian cyclopoids represent one of the richest endemic faunas of freshwater cyclopoid copepods. The genus Diacyclops Kiefer, 1927 is the most numerous by species number in the lake. In this work, molecular-phylogenetic analysis of 14 species and 1 sub-species from Lake Baikal and its water catchment basin is performed. The regions of mitochondrial cytochrom-oxydase I (COI) and of nuclear small-subunit 18S rRNA were used as evolution markers. In the obtained set of nucleotide sequences of COT gene, an effect of synonymous substitution saturation is revealed. Baikalian representatives of the genus Diacyclops form at phylogenetic schemes by two markers a monophyletic griup, it suggest their origin from a common ancestral form. Preliminary estimate of the age of this group is 20-25 My.

[Ultrastructural features of the epidermis in turbellaria from the Lake Baikal Geocentrophora wagini (Lecithoepitheliata, Plathelminthes)].

The epidermis of Geocentrophora wagini was studied using transmission electron microscopy. The turbellarian body was entirely covered by cilia, whose density was higher on the ventral surface compared with the dorsal one. In all regions examined, the epidermis was made up of a one-layered insunk epithelium. The basal matrix, underlying the epidermis, was a well developed basement membrane (BM) with bilayered structure, overlying the muscle network of circular and longitudinal fibers. The double plasma membranes, extending from the apical surface of epidermis to BM, were linked by specialized cell junctions. This suggested that epidermis had a cellular rather than a cyncytial arrangement. Each insunk epidermal cell was made of two unequal parts: a comparatively thin surface plate attached to BM by hemiadherens junctions, and a massive nucleated portion located below the body wall musculature in the parenchyma. A thin cytoplasmic bridge connected the epidermal plate with the nucleated cell body. The epidermal plates were joined by belt-like junctions along their adjacent surfaces. Inconspicuous zonula adherens (ZA) had a most apical position, and prominent septate junction was arrayed proximally to this zonula. Except ZA, cell boundaries in epidermis were frequently flanked by rows of light tubules and vesicles. In the basal half of the epithelial sheet, they were occassionally accompanied by single cisternae of rough endoplasmic reticulum (RER). The ultrastructure of the insunk cell body and that of the surface plate showed a considerable similarity. The common features were distinctive profiles of RER and GA, the presence of epitheliosomes, light tubules and vesicles, centrioles and fibrous granules. Thus, ultrastructural features allow a rather reliable identification of epidermal cells in the parenchyma, despite the absence of any visible morphological association between cell body and its epidermal plate.

[Molecular phylogeny of Turbellaria, based on data from comparing the nucleotide sequences of 18S ribosomal RNA genes]. / Molekuliarnaia filogeniia (Turbellaria), osnovannaia na dannykh sravnitel'nogo analiza nukleotidnykh posledovatel'nostei gena 18S ribosomnoi RNK.

Polymerase chain reaction and direct sequencing of the 5'-end region of the 18S ribosomal RNA gene were used to infer phylogenetic relationship among turbellarian flatworms from Lake Baikal. Representatives of 5 orders (Tricladida--10 spp., Lecithoepitheliata--5 spp., Prolecithophora--3 spp., Proseriata and Kalyptorhynchia one for each) were studied; nucleotide sequence of more than 340 nucleotides was determined for each species. Consensus sequence for each order having more than one representative species was determined. Distance matrix and maximum parsimony approaches were applied to infer phylogenies. Bootstrap procedure was used to estimate confidence limits, at the 100% level by bootstrapping, the group of three orders: Kalyptorhynchia, Proseriata and Lecithoepitheliata was found to be monophyletic. However, subsets inside the group had no significant support to be preferred or rejected. Our data do not support traditional systematics which joins two suborders Tricladida and Proseriata into the single order Seriata, and also do not support comparative anatomical data which show close relationship of Lecithoepitheliata and lower Prolecithophora.

Genetic divergence of Asiatic Bdellocephala (Turbellaria, Tricladida, Paludicola) as revealed by partial 18S rRNA gene sequence comparisons.

Polymerase chain reaction (PCR) and direct sequencing of small ribosomal RNA genes were used for analysis of genetic differences among Asiatic species of freshwater triclad genus Bdellocephala. Representatives of four species and four subspecies of this genus were used to establish homology between nucleotides in the 5'-end portion of small ribosomal RNA gene sequences. Within 552 nucleotide sites of aligned sequences compared, six variable base positions were discovered, dividing Bdellocephala into five different genotypes. Sequence data allow to distinguish two groups of these genotypes. One of them unites species from Kamchatka and Japan, another one unites Baikalian taxa. Agreement between available morphological, cytological and sequence data is discussed.

Phylogenetic relationships of Baikalian species of Prorhynchidae turbellarian worms as inferred by partial 18S rRNA gene sequence comparisons (preliminary report).

The polymerase chain reaction (PCR) and direct sequencing of RNA genes were applied for phylogenetic relationship analysis among Turbellaria (flatworms). Representatives of 10 species of the genus Geocentrophora (Prorhynchidae family) were used to establish homology between nucleotides in the 5'-end portion of the 18S rRNA gene sequences. Sequence data were subdivided into 3 subsets--unpaired and basepaired (according to rRNA secondary structure) and transversion, including only transversion substitutions--that were independently analyzed to construct phylogenetic trees. Distance matrix and maximum parsimony approaches were applied to infer phylogenies. Trees were examined in terms of morphological taxonomy. The DNA sequences support the taxonomy and systematics of Baikalian flatworms. The hypothesis of Geocentrophora origin in Baikal is discussed.