scholarly journals Global DNA Methylation Of Brain Neurons In Acute Poisoning With Clozapine And Its Combination With Alcohol: An Experimental Study

2021 ◽  
Vol 10 (3) ◽  
Author(s):  
Anastasiya S. Babkina ◽  
Maryam B. Khadzhiyeva ◽  
Irina V. Ostrova ◽  
Ivan A. Ryzhkov ◽  
Arkady M. Golubev
Keyword(s):  
Dna Methylation ◽  
Dna Fragmentation ◽  
Methylation Level ◽  
Gastric Tube ◽  
Acute Poisoning ◽  
Control Group ◽  
Global Dna Methylation ◽  
Brain Neurons ◽  
Ethanol Group ◽  
Risk Of Death

Background — Acute poisoning with atypical neuroleptic clozapine is characterized by rapid progression, high risk of death and severe neurological manifestations. Neurotoxic effects of this pharmaceutical drug have also been reported at therapeutic doses. The pathogenesis of brain damage in acute clozapine poisoning is not fully understood. Changes in DNA methylation level may play an important role in the mechanisms of drug neurotoxicity. The available data on the effect of clozapine on brain cell DNA provide a rationale for studying the epigenetic aspects of the pathogenesis of acute poisoning with this neuroleptic agent. The objective of our study was to evaluate the global DNA methylation level in rat brain neurons in acute poisoning with clozapine and its combination with ethanol. Material and methods — Clozapine – 150 mg/kg in 2.0 ml of normal saline solution, or clozapine – 150 mg/kg in 2.0 ml of 40% ethanol were administered via a gastric tube to adult male Wistar rats (n=21) under anesthesia with sevoflurane. In the control group, saline was administered via a gastric tube. Animals were euthanized four hours after drug administration. Autopsy was performed with the collection of brain samples for histochemical examination and determination of the DNA methylation level using the fluorometric method. To detect DNA in sections of paraffin-embedded tissue, we used the Feulgen staining. The TUNEL method was employed to detect DNA fragmentation. Results — An increase in the level of global DNA methylation in brain neurons was found in the clozapine and clozapine+ethanol groups. The average level of methylated DNA in the clozapine+ethanol group was higher than in the control group or clozapine group (2.56±0.31 vs. 1.35±0.1, p=0.007 and 1.70±0.33, p=0.044, respectively). An increase in the mean optical density of the cortical neuron nuclei was observed in the clozapine+ethanol group compared with the control group and clozapine group. DNA fragmentation was not detected in any experimental group. Conclusion — Acute poisoning with clozapine in combination with alcohol caused an increase in the global DNA methylation level in brain neurons, which may have played a significant role in the pathogenesis of acute clozapine poisoning and could be an important factor in the neurotoxicity of this medication.

scholarly journals Evaluation of Global DNA Methylation and Gene Expression of Izumo1 and Izumo1r in Gonads after High- and Low-Dose Radiation in Neonatal Mice

Biology ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1270
Author(s):  
Akifumi Nakata ◽  
Keisuke Sato ◽  
Yohei Fujishima ◽  
Valerie Goh Swee Ting ◽  
Kanade Nakayama ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Ionizing Radiation ◽  
Methylation Level ◽  
Low Dose ◽  
Control Group ◽  
Global Dna Methylation ◽  
Low Dose Radiation ◽  
Methylation Patterns ◽  
Dose Radiation

The intergenerational effects from chronic low-dose exposure are matters of concern. It is thus important to elucidate the radiation-induced effects of germ cell maturation, fertilization and embryonic development. It is well known that DNA methylation levels in CpG sites in gametes are reprogrammed in stages during their maturity. Furthermore, the binding of Izumo on the surface of sperm and Juno on the surface of oocytes is essential for fertilization. Thus, there is a possibility that these genes are useful indicators to evaluate fertility in mice after irradiation exposure. Therefore, in this study, we analyzed global DNA methylation patterns in the testes and gene expression of Izumo1 and Izumo1r (Juno) in the gonads of mice after neonatal acute high-dose ionizing radiation (HDR) and chronic low-dose ionizing radiation (LDR). One-week-old male and female mice were irradiated with a total dose of 4 Gy, with acute HDR at 7 days at a dose rate of 30 Gy/h and LDR continuously at a dose rate of 6 mGy/h from 7 to 35 days. Their gonads were subsequently analyzed. The results of global DNA methylation patterns in the testes showed that methylation level increased with age in the control group, the LDR group maintained its DNA methylation level, and the HDR group showed decreased DNA methylation levels with age. In the control group, the gene expression level of Izumo1 in the testis did not show age-related changes, although there was high expression at 100 days of age. However, in the LDR group, the expression level recovered after the end of irradiation, while it remained low regardless of age in the HDR group. Conversely, gene expression of Izumo1r (Izumo1 receptor) in the ovary decreased with age in the control group. Although the gene expression of Izumo1r decreased with age in the LDR group, it remained low in the HDR group. Our results indicate that LDR can induce different DNA methylation patterns, and both high- and low-dose radiation before sexual maturity might affect gametogenesis and fertility.

scholarly journals Potential effect of tobacco cigarettes smoking on global DNA methylation status and protamines transcripts in human spermatozoa

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Mohammed M. Laqqan ◽  
Maged M. Yassin
Keyword(s):  
Dna Methylation ◽  
Cigarette Smoking ◽  
Dna Fragmentation ◽  
Methylation Status ◽  
Human Spermatozoa ◽  
Global Dna Methylation ◽  
Transcription Level ◽  
Heavy Smokers ◽  
Protamine 1 ◽  
Protamine 2

Abstract Background Epigenetics refers to an alteration in gene expression without alteration in the sequence of DNA and this process may be affected by environmental factors and lifestyle like cigarette smoking. This study was designed to evaluate the potential effect of cigarette smoking on the global DNA methylation status and the transcription level of protamine 1 and protamine 2 in human spermatozoa. A total of 188 semen samples were collected from men with a mean age of 34.9 ± 5.8 years old (98 heavy smokers and 90 non-smokers). The DNA and RNA were isolated from purified spermatozoa, then the status of global DNA methylation and the transcription level of protamine 1 and protamine 2 were evaluated using ELISA and qPCR, respectively. The chromatin non-condensation and DNA fragmentation in human spermatozoa were evaluated using chromomycin A3 staining and TUNEL assay, respectively. Results A significant increase has been found in the status of global DNA methylation in spermatozoa of heavy smokers compared to non-smokers (7.69 ± 0.69 ng/μl vs. 4.90 ± 0.40 ng/μl, P < 0.001). Additionally, a significant reduction has been found in transcription level of protamine 1 (25.49 ± 0.31 vs. 23.94 ± 0.40, P < 0.001) and protamine 2 (28.27 ± 0.39 vs. 23.45 ± 0.30, P < 0.001) in heavy smokers. A downregulation has been found in the transcription level of protamine 1 and protamine 2 with a fold change of 0.497 and 0.047, respectively. A significant increase has been shown in the level of DNA fragmentation and chromatin non-condensation in heavy smokers compared to non-smokers (P < 0.001). On the other hand, a significant positive correlation has been found between sperm chromatin non-condensation, sperm DNA fragmentation, transcription level of protamine 1, transcription level of protamine 2, and global DNA methylation status (r = 0.304, P < 0.001; r = 0.399, P < 0.001; r = 0.216, P = 0.003; r = 0.494, P < 0.001, respectively). Conclusion Tobacco cigarette smoking has a potential influence on the global DNA methylation and the transcription level of protamine genes in human spermatozoa, and consequently, affect negatively on the semen parameters.

scholarly journals ANALYSIS OF THE ASSOCIATION OF THE METHYLATION LEVELS OF MIR10B AND MIR21 GENES IN BLOOD LEUKOCYTES WITH ADVANCED CAROTID ATHEROSCLEROSIS

2018 ◽  
Vol 33 (2) ◽  
pp. 77-82
Author(s):  
Iu. A. Koroleva ◽  
A. A. Zarubin ◽  
A. V. Markov ◽  
A. N. Kazancev ◽  
O. L. Barbarash ◽  
...  
Keyword(s):  
Risk Factors ◽  
Dna Methylation ◽  
Methylation Level ◽  
Carotid Atherosclerosis ◽  
Gene Promoter ◽  
Control Group ◽  
Coding Region ◽  
Blood Leukocytes ◽  
Atherosclerosis Risk Factors

Complications of atherosclerosis remain the leading cause of morbidity and mortality worldwide. MiRNAs are short regulatory molecules that are involved in all processes of pathogenesis. Expression of miRNAs is regulated by DNA methylation. Methylation and/or expression of MIR10B and MIR21 genes are known to vary in atherosclerotic tissues of the arteries, but there is no data about the changes in the methylation levels of these genes in blood leukocytes and their association with atherosclerosis risk factors.Objective.To evaluate the association of methylation levels of MIR10B and MIR21 genes in the blood leukocytes with risk factors and pathogenetically significant traits of carotid atherosclerosis.Material and Methods. DNA for the study was extracted from the samples of blood leukocytes of 122 patients with advanced carotid atherosclerosis as well as from blood leukocytes of 135 individuals in the control group. The DNA methylation level was analyzed by bisulfite pyrosequencing.Results.The methylation level of the MIR10B and MIR21 genes in leukocytes of patients with atherosclerosis is higher than in the leukocytes of the control group. In leukocytes of patients with carotid atherosclerosis the methylation level of the MIR21 gene promoter was correlated with type 2 diabetes and serum cholesterol level, and the methylation level of the coding region of the MIR10B gene was correlated with smoking.Conclusions.The level of DNA methylation in the regions of MIR10B and MIR21 genes in blood leukocytes is associated with the risk of advanced atherosclerosis of the carotid arteries. 

scholarly journals The Effect of Choline on DNA Methylation in the Hippocampus of the Offspring of Gestational Diabetes Mellitus (GDM) Mice

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1252-1252
Author(s):  
Kaydine Edwards ◽  
Xinyin Jiang ◽  
Hunter Korsmo
Keyword(s):  
Dna Methylation ◽  
Group Versus ◽  
Control Group ◽  
Global Dna Methylation ◽  
High Fat ◽  
Methylation Analysis ◽  
Feeding Condition ◽  
Control Groups ◽  
Funding Sources ◽  
Dna Methylation Analysis

Abstract Objectives The objective of this project is to determine the effects of choline supplementation on DNA methylation in the hippocampus of the offspring from mouse dams with GDM. Methods In this study, female mice were divided into four groups: high fat (HF) feeding (to induce GDM), HF with choline supplementation, normal fat (NF) control and NF with choline supplementation. The experimental groups followed their diets and supplements for 4 weeks before timed mating and throughout gestation. Thereafter, they were fed the NF diet during lactation. After weaning, the offspring were fed the HF diet for 6 weeks before dissection. Brain hippocampus was then dissected for DNA extraction and global DNA methylation analysis. Results Global DNA methylation was increased in the NF choline supplemented group versus the NF control group (P = 0.056); however, there were no differences between the HF choline versus the HF or NF control groups (P = 0.992). Conclusions In summary, there was an interaction between maternal HF feeding and choline supplementation in influencing global DNA methylation. Maternal HF feeding eliminated the increase in offspring hippocampal DNA methylation by choline supplementation observed under the NF feeding condition. Funding Sources NIGMS.

scholarly journals Repeats as global DNA methylation marker in bovine preimplantation embryos

2017 ◽  
Vol 62 (No. 2) ◽  
pp. 43-50 ◽  
Author(s):  
W. Li ◽  
A. Van Soom ◽  
L. Peelman
Keyword(s):  
Dna Methylation ◽  
Mutation Rate ◽  
Methylation Level ◽  
Methylation Status ◽  
Repetitive Sequences ◽  
Cell Types ◽  
Blastocyst Stage ◽  
Repeat Sequence ◽  
Preimplantation Embryos ◽  
Global Dna Methylation

DNA methylation undergoes dynamic changes and is a crucial part of the epigenetic regulation during mammalian early development. To determine the DNA methylation levels in bovine embryos, we applied a bisulfite sequencing based method aimed at repetitive sequences including three retrotransposons (L1_BT, BovB, and ERV1-1-I_BT) and Satellite I. A more accurate estimate of the global DNA methylation level compared to previous methods using only one repeat sequence, like Alu, could be made by calculation of the weighted arithmetic mean of multiple repetitive sequences, considering the copy number of each repetitive sequence. Satellite I and L1_BT showed significant methylation reduction at the blastocyst stage, while BovB and ERV1-1-I_BT showed no difference. The mean methylation level of the repetitive sequences during preimplantation development was the lowest at the blastocyst stage. No methylation difference was found between embryos cultured in 5% and 20% O<sub>2</sub>. Because mutations of CpGs negatively influence the calculation accuracy, we checked the mutation rate of the sequenced CpG sites. Satellite I and L1_BT showed a relatively low mutation rate (1.92 and 3.72% respectively) while that of ERV1-1-I_BT and BovB was higher (11.95 and 24% respectively). Therefore we suggest using a combination of repeats with low mutation rate, taking into account the proportion of each sequence, as a relatively quick marker for the global DNA methylation status of preimplantation stages and possibly also for other cell types.

scholarly journals Sperm global DNA methylation level: association with semen parameters and genome integrity

Andrology ◽  
2015 ◽  
Vol 3 (2) ◽  
pp. 235-240 ◽  
Author(s):  
D. Montjean ◽  
A. Zini ◽  
C. Ravel ◽  
S. Belloc ◽  
A. Dalleac ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Methylation Level ◽  
Semen Parameters ◽  
Genome Integrity ◽  
Global Dna Methylation

Changes in methylation patterns of multiple genes from peripheral blood leucocytes of Alzheimer's disease patients

2013 ◽  
Vol 25 (2) ◽  
pp. 66-76 ◽  
Author(s):  
Yaping Hou ◽  
Huayun Chen ◽  
Qiong He ◽  
Wei Jiang ◽  
Tao Luo ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Dna Methylation ◽  
Methylation Level ◽  
Target Genes ◽  
Normal Population ◽  
Cpg Islands ◽  
Methylation Pattern ◽  
Control Group ◽  
Specific Pcr

BackgroundEfforts aiming at identifying biomarkers and corresponding methods for early diagnosis of Alzheimer's disease (AD) might be the most appropriate strategy to initiate promising new treatments and/or prevention of ADObjectiveThe aim of our study is to assess the association of DNA methylation pattern of various leucocyte genes with AD pathogenesis in order to find potential biomarkers and corresponding methods for molecular diagnosis of AD.MethodsDNA methylation level of various genes in AD patients and normal population were compared by bisulphite sequencing PCR and methylation-specific PCR (MSP). Furthermore, real-time PCR was used to explore the effects of DNA methylation on the expression of target genes.ResultsResults showed significant hypermethylation of mammalian orthologue of Sir2 (SIRT1) gene in AD patients compared with normal population. Meanwhile, changes in methylation level of SIRT1 gene between different severities of AD were also found. Specific primers were designed from the SIRT1 CpG islands to differentiate AD and control group by MSP method. Besides, significant demethylation of β-amyloid precursor protein (APP) gene was observed in AD patients, whereas no difference was observed in other AD-related genes. Moreover, significant decrease in expression of SIRT1 gene and increase in expression of APP gene were also found in AD patients. In addition, the expression level of SIRT1/APP genes was associated with the severity, but not with the age or gender, of AD patients.Conclusion:SIRT1 and APP might be the interesting candidate biomarkers and valuable for clinical diagnosis or treatment of AD.

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