


Vol 55, No 6 (2019)
- Year: 2019
- Articles: 17
- URL: https://journal-vniispk.ru/1022-7954/issue/view/11863
Reviews and Theoretical Articles
Prejudices against Microsatellite Studies and How to Resist Them
Abstract
In this article, we summarize the basic concepts and techniques widely used in microsatellite studies. We describe briefly the main stages of the work and provide references for the most frequently used computational programs for processing microsatellite data. The article also gives a brief analysis of the biological meaning of the indices most often encountered in microsatellite studies.



Aldosterone Biosynthesis: Genetic Control and Contribution to the Development of Arterial Hypertension
Abstract
Data on the contribution of gene polymorphism of proteins and enzymes of aldosterone metabolism to arterial hypertension are considered. On the basis of the analysis of scientific papers (eLIBRARY.RU, PubMed) and open databases (NCBI, GeneCard, QIAGEN, and related resources), the current status of ideas about the aldosterone metabolism genes (STAR, CYP11A1, HSD3B1, CYP21A2, CYP11B1, CYP11B2, UGT2B7), their expression, polymorphism, and the effect of the genetic variability on the encoded product was assessed. The role of the balance of aldosterone precursors in the regulation of blood pressure and the mutual influence of mineralocorticoids during aldosterone biogenesis on the final product of the reaction were analyzed. The main discussions on the studied issue were noted.



Genotoxic and Carcinogenic Effects of Industrial Factors in Coal Mining and Coal-Processing Industry (Review)
Abstract
This paper presents a review of research devoted to the study of genotoxic and carcinogenic effects of exposure to factors of the production environment of the coal mining and coal processing industries. The first part of the review analyzes mutagenic effects: chromosomal aberrations (CA), sister chromatid exchanges (SCE), micronuclei (MN), and DNA comets in coal mining workers (mines) and workers in coal processing (coal heating plants, coking plants) enterprises. The second part is devoted to the study of cancer incidence in workers employed in the coal industry. The third part deals with works that carried out a comprehensive analysis of the mutagenic and carcinogenic effects in miners and workers at coal-fired power plants and coking plants. In general, the analysis of the literature confirmed the high genotoxic and carcinogenic danger of the working conditions for workers employed in the coal mining and coal processing industry.



Plant Genetics
SWEET Uniporter Gene Family Expression Profile in the Pitcher Development in the Carnivorous Plant Nepenthes sp.
Abstract
Transcriptome analysis of leaf and pitcher at different developmental stages in the carnivorous plant Nepenthes sp. identified 20 cDNAs of the SWEET gene family encoding sugar uniporters of classes I–IV. The structure of NSWEET proteins generally corresponded to the 3-1-3 scheme typical of SWEET proteins in eukaryotes. The variability in the expression of NSWEET genes in the mature leaf and in the three stages of pitcher development indicates the possible functional diversity of these genes. It has been suggested that class I transporters (NSWEET2d, 2f, and 2h) may participate in export of sugars from the leaf to the site of pitcher meristem initiation, while proteins NSWEET1, 2a, 2k (class I), 4a, 4b, 4d (II), and 12c (III) may participate in the delivery of sugars for the primary development of the pitcher. In subsequent stages, they can be replaced by NSWEET2b, 2c, 2e, 2i, 2j (class I), 4c (II), and 12b (III), delivering hexoses and sucrose to the growing pitcher. In the fully formed pitcher, proteins NSWEET12a (III), 2g (I), and 16 (IV) can export sugars from the digestive fluid to the leaf.



Divergence of Oxytropis Species from the Section Verticillares (Fabaceae) of Steppe Flora of Baikal Siberia Based on Analysis of Chloroplast DNA
Abstract
The analysis of nucleotide sequence polymorphism of chloroplast DNA psbA–trnH, trnL–trnF, and trnS–trnG intergenic spacers in the Oxytropis lanata, O. myriophylla, O. oxyphylla, O. selengensis, and O. stukovii (section Verticillares) of the steppe flora of Baikal Siberia demonstrated that the populations are characterized by a high haplotype (0.634–1.000) and relatively low (0.0003–0.0045) nucleotide diversity. Statistically significant values of genetic distances between the populations of the same species change from 0.19382 to 0.43449. Marker nucleotide substitutions and indels were detected in O. lanata, O. gracillima, O. interposita, O. pumila, and O. mongolica, indicating a significant divergence of their chloroplast genomes. A weak differentiation (ΦST = 0.137, P < 0.0001) between O. oxyphylla, O. selengensis, O. stukovii and O. bargusinensis and also low values of genetic distances (0.08939–0.32339) (corresponding to interpopulation distances), absence of differentiating molecular markers, formation of a single haplogroup in the median network, and presence of common haplotypes indicate that these species are a genetically homogeneous group probably formed as a result of relatively recent and rapid divergence from the common ancestor, as well as a high degree of interspecies hybridization.



Animal Genetics
Analysis on Microsatellite Loci of Daphnia similoides sinensis and Rapid Development of Primer Based on Transcriptome Sequencing
Abstract
In this study, we performed transcriptome sequencing of D. similoides sinensis and developing of simple sequence repeats (SSRs) markers to identify valuable markers for Daphnia molecular genetics. The total length of D. similoides sinensis transcriptome sequence was 40 093 546 bp, which contained 12 017 SSR markers and 21 344 microsatellite loci. 2164 SSR primer pairs were obtained through design for 21 344 sequences which contained the loci of SSRs. 60 primer pairs were randomly selected, 11 SSR primer pairs with polymorphism were obtained by polyacrylamide gel electrophoresis and capillary electrophoresis. A total of 88 alleles of SSRs were amplified, of which 52 belonged to polymorphic loci (the percentage of polymorphic loci was 59%). The polymorphism information content of the 11-pairs SSR primers ranged from 0.4 to 0.75, with an average of 0.58, indicating that these primers had rich polymorphism. The results suggested that the 11 primers pairs might provide important values in revealing the genetic diversity and phyletic evolution of D. similoides sinensis.



Human Genetics
The Influence of microRNAs in Regulation of Hormone Dependence in Prostate Cancer Cells
Abstract
A spectrum of differentially expressed microRNAs was determined by the massively parallel sequencing method in normal healthy prostate tissues, in hormone-dependent prostate cancer samples, and in the LNCaP and DU145 cell lines. A set of microRNAs in tumors and prostate cancer (PCa) cell lines was identified on the basis of the changes in expression compared with that in normal prostate tissues. Twenty-seven aberrantly expressed microRNAs were detected in tumor tissues and ten of them showed significant changes in expression in LNCaP and DU145 cells. Seven of them demonstrated the change of the expression in the same direction in all the tumor samples as well as in the PCa cell lines. The expression of miR-148a changed in DU145 cells in the opposite direction compared with that in LNCaP cells and tumors. The expression of let-7c, let-7b, miR-99a, miR-125b-2, miR-100, miR-10a, and miR-31 was reversed in DU145 cells compared with LNCaP cells. However, these microRNAs exhibited no significant changes in expression in tumors. It turns out that the target of miR-148a, let-7b, and microRNAs, included in the miR-99a/let-7c/miR-125b-2 cluster, the expression of which increased in LNCaP cells and decreased in DU145 cells, is the insulin-like growth factor receptor gene 1 (IGF1R). The obtained data make it possible to assume that the differences in the effect of microRNAs in cell lines are connected with their repressive influence on IGF1R expression in hormone-sensitive LNCaP cells and an absence of such influence in the hormone-independent DU145 cell line.



Association Analysis of Polymorphic Gene Variants in the JAK/STAT Signaling Pathway with Aging and Longevity
Abstract
The aim of this study was to investigate the associations between the polymorphic loci in the JAK/STAT signaling pathway and the aging processes and longevity in the populations of ethnic Tatars. For this aim, we analyzed the age dynamics of genotype frequencies of the JAK1 (32454C>T, rs310216), JAK3 (14385C>T, rs3212780), STAT1 (14228A>C, rs12693591), STAT3 (19006G>C, rs2293152), and STAT5A (1985T>C, rs9889323) genes. Using logistic regression analysis, we discovered the association of these polymorphic loci with advanced age. In the general study group, the probability of the JAK1*C/*Т genotype was higher among persons of advanced age (69–80 years old, p = 0.031, OR = 1.070). In turn, the chance to reach advanced age was lower among the carriers of the JAK1*Т/*Т (58–82 years old, p = 0.014, OR = 0.958) and STAT5A*T/*T (46–109 years old, p < 0.001, OR = 0.979) genotypes. The probability of attaining longevity was higher among men with the STAT3*С/*С genotype (25–98 years old, p = 0.027, OR = 1.016) and was lower among men with the STAT3*G/*С genotype (73–98 years old, p = 0.044, OR = 0.950). In the group of women, the probability to reach 80 years of age and over was higher among carriers of the JAK3*Т/*С (48–81 years old, p = 0.046, OR = 1.024), STAT1*С/*A (61–88 years old, p = 0.041, OR = 1.035), and STAT5A*T/*C (46–82 years old, p < 0.0001, OR = 1.044) genotypes. In turn, among middle age women with the STAT1*С/*С genotype, the probability to attain longevity was lower (55–109 years old, p = 0.013, OR = 0.980).



Study of the Role of the Main Factors of Population Dynamics in the Mechanism of Differentiation and Formation of Diversity and Genetic Load of Hereditary Diseases in Subpopulations of the Karachay-Cherkess Republic
Abstract
The generalized results of the genetic and epidemiological study of the genetic structure of various ethnic groups of the Karachay-Cherkess Republic (KChR) through various genetic systems are presented: by the genes causing AD, AR, and X-linked hereditary diseases and by nonbiological parameters estimated by the methods of population statistics (random inbreeding FST, Malécot’s isolation-by-distance parameters, migration index, and Crow index and its components). The total size of the investigated population was 410 367 individuals (city of Cherkessk, Ust-Dzhegutinsky, Karachayevsky, Malokarachayevsky, Prikubansky, Khabezsky, Nogaysky, Adyge-Khablsky, Urupsky, and Zelenchuksky districts). On the basis of the study of correlations between different characteristics of the population genetic structure, it is assumed that the main cause of genetic differentiation of KChR subpopulations in terms of diversity and load of hereditary diseases (AD, AR, and X-linked) is genetic drift and migration under decreased influence of natural selection and mutational process.



Effect of the ZNF804A Gene and Obstetrical Complications on Clinical Characteristics of Schizophrenia
Abstract
To explore for the first time an effect of gene × environment (G × E) interactions on the clinical characteristics of schizophrenia, we studied the ZNF804A rs1344706 polymorphism, a genome-wide supported risk variant of psychosis, and obstetrical complications (OC) as a risk-modifying factor for psychiatric disorders, in 369 patients with schizophrenia (203 women, mean age 29.7 ± 10.1 years, age at disease onset 21.6 ± 7.3 years). A history of at least one definite OC was present in 111 patients, while 258 patients had no history of OC. Clinical characteristics, including age at disease onset and severity of symptoms assessed with PANSS, were compared by the ZNF804A genotype in the groups with and without OC. Patients with the risk AA genotype and OC had higher scores on the subscale of general psychopathological symptoms compared to the carriers of the CC genotype without OC (p = 0.007). The results demonstrated an additive effect of ZNF804A rs1344706 and OC on symptoms severity in schizophrenia.



A Study of the Association between Breastfeeding and DNA Methylation in Peripheral Blood Cells of Infants
Abstract
This article reports on new data on the association of breastfeeding with DNA methylation in the peripheral blood cells of 37 children aged from 9 months to four years. Whole-genome DNA methylation profiling was performed using the Illumina Methylation EPIC array. The Epigenome-Wide Association Study (EWAS) revealed an association between the duration of breastfeeding and the methylation level of 4276 СpG sites related to 2635 genes. According to the functional annotation, these genes were predominantly involved in the control of cell signaling systems, the development of anatomical structures and cells, and, above all, were related to the development and function of the immune system and the CNS. The results of the study allowed assuming a special role of the oxytocin signaling pathway, as a potential trigger of coordinated epigenetic changes in the genes involved in the CNS function in response to breastfeeding.



Analysis of Two Polymorphic Repeats IVS3 Poly A and IVS10 CA in Tunisian Cystic Fibrosis Patients: Case–Control Study
Abstract
The aim of this study was to determine a possible association of IVS3 poly A and IVS10 CA microsatellites with CF in case–control Tunisian groups and to compare the results with the findings in Italian population. Both microsatellites were analyzed by capillary electrophoresis in 70 normal subjects and 60 Tunisian CF patients. The statistical distribution of IVS3 poly A and IVS10 CA polymorphism shows a significant difference between CF patients and controls group. 12 different alleles were found for IVS3 poly A which only five were identified in CF patients. IVS3 poly (A)19 was the most prevalent allele in CF patients with 75%. Nevertheless, IVS3 poly (A)18 was more common in normal individuals (27.14%). Seven different alleles were found for IVS10 CA. The IVS10 (CA)16 was the most prevalent allele both in normal individuals and CF patients with 30.71 and 52.5% frequencies respectively. Concerning the association of these two markers with F508del mutation, we noted that IVS3 poly (A)19 is the most common in our CF patients but IVS3 poly (A)18 is specific to the CF Italian patients. Furthermore, IVS10 (CA)16 and IVS10 (CA)19 are the most dominant (29.16%) in our samples and IVS10 (CA)23 is specific to the CF Italian subjects (94.4%). Our study allowed us to identify the haplotype distribution of both markers IVS3 poly A and IVS10 CA in CF and control subjects. The results obtained show a strong association between the most frequent F508del mutation and IVS3 poly (A)19, IVS10 (CA)16 and IVS10 (CA)19.



Short Communications
Genetic Differentiation of Rhododendron aureum Georgi at Nuclear Microsatellite Loci
Abstract
SSR analysis of Rhododendron aureum Georgi in eight populations (63 samples) from Siberia (West and East Sayans) and Kamchatka was performed. Polymorphism of five SSR loci showed a high level of genetic diversity in all the populations (mean values Ho = 0.667, He = 0.638). The analysis of molecular variation (AMOVA) revealed that about 16% of the genetic diversity of R. aureum occurs in interpopulation differences (Fst = 0.161, p = 0.001). The Bayesian approach in the STRUCTURE program revealed that the species has a heterogeneous genetic structure—populations from Kamchatka form a separate cluster and show high homogeneity, and populations from Western and Eastern Siberia are of mixed origin. The obtained results indicate the prospects of using this type of markers for population-genetic studies of R. aureum.



The Frequencies of Alleles of Single Nucleotide Substitutions in the CCK and CCK2R Genes in Some Russian Cattle Breeds
Abstract
Allele frequencies of three substitutions in the CCK (rs42891945 and rs42891946) and CCKBR (rs42670352) genes were identified in three Russian cattle breeds: Holstein (dairy), Bestuzhev (dual-purpose), and Kalmyk (beef). The most promising for further associative studies is the rs42670352 substitution in the CCKBR gene.



Microsatellite Variability of the Arctic Rainbow Smelt Osmerusdentex from the White Sea
Abstract
We studied the genetic variability of ten microsatellite loci in Arctic rainbow smelt datasets from Kandalaksha and Mezen bays of the White Sea. Average estimates of expected heterozygosity ranged from 0.116 to 0.640 at different loci, with an average of 0.407. Smelt from Kandalaksha Bay is characterized by lower values of genetic diversity (HE = 0.365–0.369, AR = 3.1–3.15) compared to dataset from Mezen Bay (HE = 0.486, AR = 4.02). Significant genetic divergence between the smelt of Kandalaksha and Mezen bays is shown; the overall estimate of genetic differentiation θ = 7.8% with 95% confidence bootstrap interval [2.8–13.6%].



Meiotic Chromosomes, Synaptonemal Complexes in a Female Viviparous Lizard (Zootoca vivipara) in Prophase I of Meiosis
Abstract
In the females of the viviparous lizard Zootoca vivipara (Lichtenstein, 1823) (family Lacertidae) from Northwest Russia (2n = 35: 32A (acrocentric autosomes) + Z1Z2W sex chromosomes), the ovarian lumen germinal vesicles (oocytes), as well as germinal lamina cells, were examined. Chromosome preparations were obtained using the direct method and the method of total oocyte nuclei spreading developed by Dresser and Moses. Chromosome preparations were stained with Giemsa; for visualization of synaptonemal complexes (SCs), total preparations of oocyte nuclei were stained with silver nitrate and DAPI. It was demonstrated that, during oogenesis in the female, primary follicles enter the early stages of the meiotic prophase I (stages from leptotene to diplotene, lampbrush chromosomes are formed). Here, for the first time, total oocyte spreads were obtained and studied. On the basis of light microscopic analysis of the oocyte SCs and taking into account their length in a female with 2n = 35, the SC karyotype is presented, consisting of 19 SC elements, among which 16 SC autosomal bivalents are distinguished. The remaining three SC elements, according to the authors, can be univalents of Z1Z2W sex chromosomes or one WZ1 bivalent and Z2 and B chromosome univalents. As in the SC karyotype of Z. vivipara males, in a female, specific features in the morphology of SC elements of chromosomes 5 and 6 were observed.



Structure and Variability of the Pax3 and EDNRB Genes in American Mink (Neovison vison)
Abstract
The Pax3 protein is a transcription factor responsible for process of differentiation neural crest stem cells during the embryonic period, in individual life Pax3 gene expression is an element of skin cells response to ultraviolet radiation (UV). Mutations of Pax3 gene are possible cause of some color patterns Quarter Horse breed, additionally some variations of Pax3 gene can disturb the process of embryonic or fetal development. Endothelin Receptor Type B (EDNRB) is another candidate gene that can affect on the distribution of the dye in skin or hairs, exons 1, 3, 4 and 5 were sequenced, any nucleotide variations were not observed. Exons 2 and 3 of the Pax3 gene along with the flanking regions were sequenced, in which a few variations were found, some of them were genotyped by means of PCR-RFLP and ASA-PCR. Statistically significant differences between the frequency of examined alleles in the various coat color groups were observed.


