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Kharzinova V.R.,Federal Agency of Scientific Organizations | Gladyr E.A.,Federal Agency of Scientific Organizations | Fedorov V.I.,Federal Agency of Scientific Organizations | Romanenko T.M.,Federal Agency of Scientific Organizations | And 4 more authors.
Sel'skokhozyaistvennaya Biologiya | Year: 2015

Reindeer (Rangifer tamndus), the only member of the genus Rangifer, is one of the most interesting object to investigate genetic diversity. One of the technique of studying the genetic structure of populations and parentage identification is to create panels of STR (short tandem repeats) markers. The aim of the current study was the development of multiplex panel of STR markers and assessment of its application to assign the parents and to study biodiversity of Russian reindeer populations. As a biological material for research we used tissue samples (part of ear's lobes) of reindeer of Even (EVN, n = 44), Evenk (EVK, n = 44), Nenets (n = 45) breeds and Tyva population (TUV, n = 35). DNA extraction was performed using Nexttec columns (Germany) according to the manufacturer's instructions. Polymorphism of nine STR markers (NVHRT76, RT9, NVHRT24, RT30, RT1, RT6, RT27, NVHRT21 and RT7) was determined by own procedures using ABI 3130×1 DNA analyzer («Applied Biosystems», USA). Statistical analysis was performed in MS Excel 2007 with the plugin GenAIEx v. 6.5, software MSA 4.05, PHYLIP, v. 3.5c, Treev32 and Structure, v. 2.3.4. The studied populations of reindeer were characterized by relatively high levels of genetic diversity. The average number of alleles per locus was 6.11±0.56 in TUV, 6.67±0.50 in NEN, 8.00±0.76 in EVN and 8.89±0.65 in EVK. The smallest effective number of alleles per locus was detected in TUV (3.37±0.47), the maximal value was in EVK (4.89±0.46 alleles per locus), and EVN and NEN occupied an intermediate position (4.42±0.53 and 3.90±0.38, respectively). The number of alleles in single loci ranged from four in NVHRT21 and NVHRT24 for TUV to twelve in RT7 for EVK and RT1 for EVN. The probability of matching genotypes (PI) for the nine loci ranged from 1.84×10-9 in NEN to 5.9×10-11 in EVK, showing the high power of the proposed marker panel for parentage identification. The calculation of the mean values of similarity coefficient Q in the ith cluster with the most probable number of clusters such as k = 3 and k = 4 (Qi/k) revealed high heterogeneity of genetic structure of studied populations. The highest degree of genetic differentiation was shown for TUV (Q2/3 = 0.899±0.034, Q3/4 = 0.883±0.035) and for NEN (Q3/3 = 0.885±0.031, Q4/4 = 0.813±0.038). The EVN and EVK population were close to each other, and a clear clustering between them was not observed. An estimation of Rst (AMOVA) showed that 11.4 % of the total molecular variability was caused by differences between populations, and 88.6 % was due to individual differences between animals (p < 0.01). Evaluation of degree of genetic differentiation of studied populations, using as criteria the values of Nei' genetic distances and pairwise comparisons of Fst showed similar trends. TUV population was the most distinct comparing to other populations (DNei = 0,283-0,502, Fst = 0,299-0,452), while it was the most differ from NEN and the closest to EVN. The minimal genetic differences were observed between EVN and EVK (DNei = 0,068, Fst = 0,032). The results show high functional power of the developed STR panel to identify the parentage and to study biodiversity in Russian reindeer populations.


Rola-Luszczak M.,National Veterinary Research Institute | Pluta A.,National Veterinary Research Institute | Olech M.,National Veterinary Research Institute | Donnik I.,Urals State Scientific Research Institute of Veterinary Medicine | And 5 more authors.
PLoS ONE | Year: 2013

Recent studies have shown that bovine leukemia virus (BLV) sequences can be classified into seven distinct genotypes based on full gp51 sequence. This classification was based on available sequence data that mainly represented the BLV population that is circulating in cattle from the US and South America. In order to aid with a global perspective inclusion of data from Eastern Europe is required. In this study we examined 44 BLV isolates from different geographical regions of Poland, Belarus, Ukraine, and Russia. Phylogenetic analysis based on a 444bp fragment of env gene revealed that most of isolates belonged to genotypes 4 and 7. Furthermore, we confirmed the existence of a new genotype, genotype 8, which was highly supported by phylogenetic computations. A significant number of amino acid substitutions were found in the sequences of the studied Eastern European isolates, of which 71% have not been described previously. The substitutions encompassed mainly the C-part of the CD4+ epitope, zinc binding peptide region, CD8+ T cell epitope, and overlapping linear epitope E. These observations highlight the use of sequence data to both elucidate phylogenetic relationships and the potential effect on serological detection of geographically diverse isolates. © 2013 Rola-Łuszczak et al.


Lebedeva I.Y.,All Russian Research Institute of Animal Breeding | Singina G.N.,All Russian Research Institute of Animal Breeding | Taradajnic T.E.,All Russian Research Institute of Animal Breeding | Zinovieva N.A.,All Russian Research Institute of Animal Breeding
Cell and Tissue Biology | Year: 2011

The mechanisms of prolactin signal transduction in generative and somatic cells of mammalian ovarian follicles are poorly understood. In this work, participation of tyrosine kinases and protein kinase C in mediation of the previously revealed modulating effects of prolactin on the nuclear maturation of bovine oocytes and the morphologic and functional state of surrounding cumulus cells in vitro has been investigated. It was found that a tyrosine kinase inhibitor genistein suppresses the stimulating action of prolactin on the completion of oocyte nuclear maturation and cumulus expansion, whereas a protein kinase C inhibitor calpostin C does not affect the hormonal effect. Furthermore, both genistein and calpostin C inhibited the inducing influence of prolactin on the proliferative activity of cumulus cells. At the same time, the retarding action of prolactin on destructive processes in cumulus cells was blocked only in the presence of calpostin C. These results show that the stimulating influence of prolactin on oocyte nuclear maturation accompanied by cumulus expansion is achieved with participation of tyrosine kinases, whereas the modulating action of the hormone on the functional state of cumulus cells depends on activation both of tyrosine kinases and protein kinase C. © 2011 Pleiades Publishing, Ltd.


Yu Lebedeva I.,All Russian Research Institute of Animal Breeding | Singina G.N.,All Russian Research Institute of Animal Breeding | Taradajnic T.E.,All Russian Research Institute of Animal Breeding | Zinovieva N.A.,All Russian Research Institute of Animal Breeding
Tsitologiya | Year: 2011

Mechanisms of prolactin signal transduction in generative and somatic cells of mammalian ovarian follicles have been studied only to a small extent. In the present work, the involvement of tyrosine kinases and protein kinase C in mediating of the previously revealed modulating effects of prolactin on the nuclear maturation of bovine oocytes and the morphologic-functional state of surrounding cumulus cells was investigated in vitro. Tyrosine kinase inhibitor, genistein, was found to suppress the stimulating action of prolactin on the completion of oocyte nuclear maturation and cumulus expansion, whereas protein kinase C inhibitor, calpostin C, did not affect these hormonal effects. Furthermore, both genistein and calpostin C inhibited the inducing influence of prolactin on the proliferative activity of cumulus cells. At the same time the retarding action of prolactin on destructive processes in cumulus cells was blocked only in the presence of calpostin C. The results of the study suggest that the stimulatory influence of prolactin on oocyte nuclear maturation and attendant cumulus expansion is achieved with the participation of tyrosine kinases, whereas the modulating action of the hormone on the functional state of cumulus cells depends on activation of not only tyrosine kinases, but also protein kinase C.

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