Functional study of potential regulatory SNPs

Arina Degtyareva1, Elena Leberfarb2, Ilya Brusentsov3, Tatiana Kuzina4, Tatiana Merkulova51ICG SB RAS, degtyareva_rso@mail.ru2ICG SB RAS, lieberfarb@bionet.nsc.ru3ICG SB RAS, brusentsovi@gmail.com4NSU, t.kuzina@g.nsu.ru5ICG SB RAS, merkulova@bionet.nsc.ru In a previous study using a new bioinformatic approach based on Chip-Seq data with antibodies to various transcription factors , histone markers , and ChiA-PET and RNA-seq data, 32 potentially regulatory single -nucleotide polymorphisms (SNPs) were associated with colorectal cancer . In this study, a functional analysis of 6 of these polymorphisms was performed using EMSA and the luciferase reporter system. These nucleotide substitutions have been shown to alter the binding of oligonucleotides to nuclear extract proteins and alter the expression of the reporter gene.

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The rs12255372 and rs7903146 polymorphisms of the TCF7L2 gene among Buryats and Russians of Eastern Siberia.

Poster (download) Ludmila Tabikhanova1, Ludmila Osipova2, Tatiana Churkina3, Daria Lichman4, Elena Voronina5, Maxim Filipenko61Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Science, Novosibirsk State University, tabikhan@bionet.nsc.ru2Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Science, Novosibirsk State University, ludos77@yandex.ru3Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Science, Novosibirsk State University, tan646464@gmail.com4Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Science, Novosibirsk State University, daria.lichman@gmail.com5Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk State University, voronina_l@mail.ru6Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk State University, max@niboch.nsc.ru The objective of the present study was to investigate the polymorphism of metabolism genes in indigenous populations of Siberia. To this end, we have studied the ethnic features of allele frequency distribution for polymorphic variants in gene TCF7L2 G103894T (rs12255372) and C53341T (rs7903146) in the samples taken from Buryats, and Russians of Eastern Siberia, and compared it with data on world populations. Samples of Eastern (N = 132) and Western (N = 278) Buryats, Russians (N = 122) and persons of mixed Buryat-Russian origin (N = 56) were genotyped by real-time PCR using competitive TaqMan- probes. Compared to the Russians, the Buryats showed a statistically significantly lower incidence of the TCF7L2 103894T and TCF7L2 53341T alleles associated with metabolic diseases and Type 2 diabetes mellitus (T2DM). Buryats are intermediate between Caucasian and East Asian populations. This agrees with the lower susceptibility of Buryats to metabolic disorders compared to the Caucasian population described in the literature. Intermediate frequencies alleles in the group of mixed origin may indicate a higher risk of the associated metabolic disorders in the descendants of mixed marriages compared to the Buryats.

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The effect of lithium carbonate on angiogenesis of hepatocellular carcinoma-29

Viktoriia Makarova1, Nataliya Bgatova2, Iuliia Taskaeva31Laboratory of ultrastructural research, Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, shedina_vika@mail.ru2Laboratory of ultrastructural research, Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, n_bgatova@ngs.ru3Laboratory of ultrastructural research, Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, inabrite@yandex.ru Гепатоцеллюлярная карцинома является одной из самых агрессивных опухолей человека с высокой распространенностью и смертностью. В то же время существует проблема устойчивости ГЦК к стандартной терапии. Поиск лекарств, блокирующих рост и метастазирование опухоли, является актуальным. Имеются данные о противоопухолевых свойствах карбоната лития. Целью нашего исследования было определить влияние лития на развитие и структуру кровеносных сосудов экспериментальной гепатоцеллюлярной карциномы-29 (G-29). Исследование было выполнено на самцах мышей CBA. Клетки гепатокарциномы-29 использовали для индукции опухолевого процесса. Изучали экспрессию эндотелиального маркера кровеносных сосудов (CD-31) и ультраструктурную организацию опухолевых сосудов эндотелиальных клеток. Было показано, что карбонат лития уменьшает объемную плотность кровеносных сосудов G-29. Ультраструктурный анализ опухолевых сосудов выявил нетипичный характер их строения. Стенки сосудов были выстланы как эндотелиальными, так и опухолевыми клетками.

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Human retinal photoreceptor cells in glaucoma: destructive changes of mithochondria and mitophagy

Poster (download) Natalia Obanina1, Nataliya Bgatova2, Valerii Chernykh3, Aleksandr Trunov4, Alena Eremina5, Vladimir Konenkov61Laboratory of ultrastructural research, Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences; Section of cytology and genetics, Department of Natural Sciences, Novosibirsk State University, n.obanina@g.nsu.ru2Laboratory of ultrastructural research, Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, n_bgatova@ngs.ru3The academician S.N. Fyodorov Federal State Institution “Intersectoral Research and Technology Complex “Eye Microsurgery”, Ministry of Healthcare of Russian Federation, Novosibirsk Branch, rimma@mntk.nsk.ru4The academician S.N. Fyodorov Federal State Institution “Intersectoral Research and Technology Complex “Eye Microsurgery”, Ministry of Healthcare of Russian Federation, Novosibirsk Branch, trunov1963@yandex.ru5The academician S.N. Fyodorov Federal State Institution “Intersectoral Research and Technology Complex “Eye Microsurgery”, Ministry of Healthcare of Russian Federation, sci@mntk.nsk.ru6Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, vikonenkov@gmail.com The leading place in the development of low vision and blindness worldwide is occupied by glaucoma. This disease is non-infectious in nature, accompanied by an increase in intraocular pressure and degeneration of neurons of the retina, optic nerve, as well as its spread in the central part of the visual analyzer. The aim of the study was to identify the characteristics of the ultrastructure of human retinal photoreceptor cells in glaucoma. The study was carried out using transmission electron microscopy and morphometry. The volume densities (VV) of mitochondria and autophagosomes containing mitochondria were determined. Disorders of orientation and swelling of the membrane discs of rod neurons in the outer segments were revealed. It is shown that sticks are more prone to destructive changes than cones.

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The expression of Bcl-2 family proteins in liver cells of 57Bl/6 mice under conditions of functional pinealectomy

Poster (download) Svetlana Viktorovna Michurina1, Irina Yurievna Ishchenko2, Sergey Alekseevich Arkhipov3, Andrey Yurievich Letyagin4, Maxim Aleksandrovich Korolev5, Evgenii Leonidovich Zavjalov6 1Group of experimental pharmacology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, s.michurina@ngs.ru 2Group of experimental pharmacology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, irenisch@mail.ru 3Group of experimental pharmacology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, arhipowsergei@yandex.ru 4Laboratory of pharmaceutical technologies SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, letyagin-andrey@yandex.ru 5Laboratory of connective tissue pathology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, kormax@bk.ru 6Center of genetis resources of laboratory animals ICG SB RAS Novosibirsk, Russia, zavjalov@bionet.nsc.ru The expression of antiapoptotic Bcl-2 protein and the proapoptotic Bad protein in liver cells of C57BL/6 mice males kept 14 days under 24-hour lighting (24hL) (photoperiod light/dark 24/0 hours) was studied. Control animals kept under standard lighting conditions (photoperiod light/dark 14/10 hours). The immunohistochemical analysis (indirect avidin-biotin-streptavidin method) and morphometric assessment were used. We found in the liver cells of mice under round-the-clock lighting the increase in the area of Bcl-2 protein expression without changing the area of Bad protein expression, that led to a decrease in the ratio of Bcl-2/Bad. This result indicated weakening of the antiapoptotic protection of organ cells and creates conditions for activation of the “mitochondrial branch” of apoptosis in animal liver cells with functional pinealectomy.

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Genetic Polymorphism Associated with Infectious Pulmonary Diseases in Siberian Populations and Among Patients with Community Acquired Pneumonia

Poster (download) Svetlama V. Mikhailova11ICG SB RAS, mikhail@bionet.nsc.ru Innate immune system is the first to respond to pathogen invasion; it activates adaptive immunity and regulates the intensity of inflammation. Two single nucleotide polymorphisms of the innate immunity genes: rs5743708 of the TLR2 and rs8177374 of the TIRAP were shown to be associated with pneumonia and tuberculosis, but the data are contradictory in different ethnic groups. We assessed the prevalence of these variants in Caucasoid and Asian population samples, and among patients with community acquired pneumonia (CAP). Carriage of the rs5743708 A allele was found to be predisposed to severe CAP (OR=2.77, p=0.021), GG/CT genotype for rs5743708 / rs8177374 is proved to be protective against it (OR=0.478, p=0.022) in Caucasoid patients. Both Caucasoid and Asian studied populations were turn to be different from European and neighboring Asian populations on the genotypes prevalence. Differentiation of CAP by the causative pathogen could help to eliminate the currently observed contradictions between different research groups.

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Study of serotonin transporter gene polymorphism Stin2 in two Siberian indigenous populations

Poster (download) Mira Khantemirova1, Daria Lichman2, Daria Bazovkina3, Anatoly Bragin4, Vladimir Naumenko5, Ludmila Osipova61Laboratory of populational ethnogenetics, Institute of cytology and genetics; Laboratory of computer genomics, Novosibirsk State University Novosibirsk, Russia, hantemiramira@mail.ru2Laboratory of populational ethnogenetics, Institute of cytology and genetics; Laboratory of computer genomics, Novosibirsk State University Novosibirsk, Russia, daria.lichman@gmail.com3Laboratory of behavioral neurogenomics, Institute of cytology and genetics Novosibirsk, Russia, daryabazovkina@gmail.com4Laboratory of computer genomics, Novosibirsk State University Novosibirsk, Russia, ibragim@bionet.nsc.ru5Laboratory of behavioral neurogenomics, Institute of cytology and genetics Novosibirsk, Russia, naumenko2002@bionet.nsc.ru6Laboratory of populational ethnogenetics, Institute of cytology and genetics; Laboratory of computer genomics, Novosibirsk State University Novosibirsk, Russia, ludos77@yandex.ru Serotonin (5-HT) participates in regulation of emotions, mood, sleep, appetite, and cognitive processes. Serotonin transporter (5-HTT, SERT) modulates serotoninergic neurotransmission through reuptake of 5-HT from the synaptic cleft. A variable number tandem repeat polymorphism found in intron 2 of the 5-HTT gene (Stin2 polymorphism) influences 5-HTT expression and is associated with anxiety, depression, suicidal behavior, obsessive-compulsive disorder. The aim of the present study was to investigate Stin2 allele frequencies in populations of Tundra Nenets and Nganasans inhabiting north and east Siberia. The frequency of the STin2.10 allele was 72.1% in Tundra Nenets and 89.4% in Nganasans. The data obtained are consistent with the uniform geographic gradient of STin2 alleles, with low frequency of the STin2.10 allele in East Asia and its increase towards west.

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Mitochondrial dysfunction and redox balance alterations in the development of AD-like pathology in OXYS rats

Poster (download) Mikhail Tyumentsev1, Natalia Muraleva2, Yulia Polienko3, Artyom Gorodetsky4, Elena Bagryanskaya51ICG SB RAS, landselur@bionet.nsc.ru2ICG SB RAS, Myraleva@bionet.nsc.ru3NIOCH SB RAS, polienko@nioch.nsc.ru4NIOCH SB RAS, gorodaa@nioch.nsc.ru5NIOCH SB RAS, egbagryanskaya@nioch.nsc.ru This study focuses on the relationship between mitochondrial dysfunction and redox-status in the context of the development of signs of Alzheimer’s disease (AD). Mitochondrial dysfunction is considered the missing link between brain aging and AD [1], the most common type of age-related dementia worldwide, but the exact causal relationship between mitochondrial dysfunction and the transition from healthy aging to AD remains to be fully understood. Oxidative stress is thought to play a significant role in this mitochondrial dysfunction, leading to cellular damage in redox imbalance. However, the extent of these processes and timing of their occurrence within the scope of AD remain hard to study, especially so in the early, pre-clinical stages of the disease. We explored the mechanisms underlying the disruption of mitochondrial function, their impact on the initiation and progression of pathological molecular cascades of AD, and assessed the changes in redox status as one of the main consequences of oxidative stress. This investigation was conducted using senescence-accelerated OXYS rats, which spontaneously develop all major signs of AD and largely reproduce the stages of the disease. We concluded that mitochondrial dysfunction appears to mediate or possibly even initiate AD-like pathology in OXYS rats. Importantly this takes place with no apparent connection to redox imbalance as on both transcriptional and biochemical levels OXYS rats display no significant changes in redox-status and ROS production as compared to controls.

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Calcium Signaling Dynamics in Single Platelets during Optically-induced Activation.

Poster (download) Darya V. Spiryova1, Alexander E. Moskalensky2, Alexei Vorob’ev3 1NSU, gerd.raz@yandex.ru 2NSU, a.mosk@nsu.ru 3NSU, vor@nioch.nsc.ru Activation of blood platelets is the main process in normal hemostasis. Unfortunately, it also makes a great contribution to the development of cardiovascular diseases. In this work, we study activation dynamics using cytosolic calcium probe. To increase the accuracy of dynamic measurements, we present a method for optical activation of single platelets in suspension. It is achieved by the use of photolabile caged activation agonists. Futhermore, a mathematical model was developed that allows to detect individual calcium peaks andВ derive their dynamic parameters.

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LYVE-1 expression in liver cells of mice with functional pinealectomy

Poster (download) Irina Yurievna Ishchenko1, Svetlana Viktorovna Michurina2, Sergey Alekseevich Arkhipov3, Andrey Yurievich Letyagin4, Maxim Aleksandrovich Korolev5, Evgenii Leonidovich Zavjalov61Group of experimental pharmacology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, irenisch@mail.ru2Group of experimental pharmacology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, s.michurina@ngs.ru3Group of experimental pharmacology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, arhipowsergei@yandex.ru4Laboratory of pharmaceutical technologies SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, letyagin-andrey@yandex.ru5Laboratory of connective tissue pathology SRICEL – a branch of ICG SB RAS Novosibirsk, Russia, kormax@bk.ru6Center of genetis resources of laboratory animals ICG SB RAS Novosibirsk, Russia, zavjalov@bionet.nsc.ru The expression of the lymphatic vessel endothelial receptor-1 hyaluronan-1 (LYVE-1) was studied in the liver cells of male C57BL / 6 mice, which were kept for 14 days under 24-hour illumination (mice with functional pinealectomy). Used immunohistochemical analysis (indirect avidin-biotin-ABC-peroxidase method) and morphometric evaluation. A 2-fold decrease in the LIVE-1 expression area was detected against a 2% increase in the optical density of LIVE-1 staining in mouse liver cells after chronic continuous illumination. Weak expression of LYVE-1 on the membranes of endothelial cells of the sinusoids of the liver may indicate a violation of the functioning of the fields of fenestration of these cells. This can lead to a decrease in the endocytotic activity of the latter, difficulty in the exchange of hematotissue tissues, deterioration of lymphatic drainage and the development of tissue hypoxia in the liver of mice with functional pinealectomy.

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