Neuromodulation: Action Potential Modeling

There have been many different studies performed in order to examine various properties of neurons. One of the most important properties of neurons is an ability to originate and propagate action potential. The action potential is a source of communication between different neural structures located in different anatomical regions. Many different studies use modeling to describe the action potential and its properties. These models mathematically describe physical properties of neurons and analyze and explain biological and electrochemical processes such as action potential initiation and propagation. Therefore, one of the most important functions of neurons is an ability to provide communication between different neural structures located in different anatomical regions. This is achieved by transmitting electrical signals from one part of the body to another. For example, neurons transmit signals from the brain to the motor neurons (efferent neurons) and from body tissues back to the brain (afferent neurons). This communication process is extremely important for a being to function properly. One of the most valuable studies in neuroscience was conducted by Alan Hodgkin and Andrew Huxley. In their work, Alan Hodgkin and Andrew Huxley used a giant squid axon to create a mathematical model which analyzes and explains the ionic mechanisms underlying the initiation and propagation of action potentials. They received the 1963 Nobel Prize in Physiology/Medicine for their valuable contribution to medical science. The Hodgkin and Huxley model is a mathematical model that describes how the action potential is initiated and how it propagates in a neuron. It is a set of nonlinear ordinary differential equations that approximates the electrical characteristics of excitable cells such as neurons and cardiomyocytes. This work focuses on modeling the Hodgkin and Huxley model using MATLAB extension - Simulink. This tool provides a graphical editor, customizable block libraries, and solvers for modeling and simulating dynamic systems. Simulink model is used to describe the mechanisms and underlying processes involved in action potential initiation and propagation

Detection of modified forms of cytosine using sensitive immunohistochemistry

Methylation of cytosine bases (5-methylcytosine, 5mC) occurring in vertebrate genomes is usually associated with transcriptional silencing. 5-hydroxylmethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC) are the recently discovered modified cytosine bases produced by enzymatic oxidation of 5mC, whose biological functions remain relatively obscure. A number of approaches ranging from biochemical to antibody based techniques have been employed to study the genomic distribution and global content of these modifications in various biological systems. Although some of these approaches can be useful for quantitative assessment of these modified forms of 5mC, most of these methods do not provide any spatial information regarding the distribution of these DNA modifications in different cell types, required for correct understanding of their functional roles. Here we present a highly sensitive method for immunochemical detection of the modified forms of cytosine. This method permits co-detection of these epigenetic marks with protein lineage markers and can be employed to study their nuclear localization, thus, contributing to deciphering their potential biological roles in different experimental contexts

5-Carboxylcytosine is localized to euchromatic regions in the nuclei of follicular cells in axolotl ovary

5-Methylcytosine (5-mC) is an epigenetic modification associated with gene repression. Recent studies demonstrated that 5-mC can be enzymatically oxidised into 5-hydroxymethylcytosine and further into 5-formylcytosine (5-fC) and 5-carboxylcytsine (5-caC). 5-caC has been found in embryonic stem cells and in mouse pre-implantation embryos but no detectable levels of this modification have been reported for somatic tissues to date. Whereas it has been suggested that 5-caC can serve as an intermediate in the process of active demethylation, the function of this form of modified cytosine remains obscure. Here we show that 5-caC is immunochemically detectable in somatic cells of axolotl ovary. We demonstrate that both 5-hmC and 5-caC are localized to the euchromatin in the nuclei of axolotl follicular cells with similar patterns of spatial distribution. Our results suggest that 5-carboxylcytosine may play a distinct functional role in certain biological contexts

Diastolic stress test in assessing functional state of postinfarction myocardium

Taking into account the conflicting literature data and the ambiguity in the interpretation of parameters of diastolic heart function in patients with coronary heart disease, we conducted an analysis of diastolic heart function in patients after myocardial infarction at the outpatient rehabilitation stage. Aim of the study was to investigate the diagnostic information content and the feasibility of diastolic stress testing to assess the functional condition of postinfarction myocardium. Material and methods. 86 patients were examined at the outpatient stage of rehabilitation 6 weeks after myocardial infarction with coronary artery stenting ad hoc. The control group consisted of 10 healthy volunteers. Structural and functional examination of the heart was performed at rest and immediately after exercise using stress echocardiography. Results and discussion. Patients after myocardial infarction have a larger indexed volume of the left atrium (30.71 ± 8.88 vs. 20.49 ± 4.04 ml/m2) and an E/e` ratio (8.45 ± 3.27 vs. 6.46 ± 1.42) in comparison with the control group. 38 (62.3 %) patients with normal left ventricular (LV) ejection fraction (EF) had unimpaired LV diastolic function, 19 (31.1 %) and 2 (3.3 %) patients had type 1 and type 2 diastolic dysfunction, respectively, 2 patients (3,3 %) had an indeterminate result. Patients with reduced LV EF have a significantly lower early diastolic mitral annular velocity (e`). The diastolic stress test revealed a significant post-exercise increase in E/e` in only one patient (from 8.92 to 18.37), who also had an initially reduced EF (32 %). The stress test showed no significant changes in diastolic heart parameters after exercise in patients suffering from heart failure with preserved EF, which may indicate relatively good diastolic reserves of the heart. Conclusions. Myocardial infarction is accompanied by the presence of LV diastolic dysfunction in 53,5 % of the patients at the 6th week of the rehabilitation period. The diastolic stress test is accompanied by a rare occurrence of stress-induced diastolic dysfunction (4 %) in patients with reduced LV EF after myocardial infarction

Semi-quantitative immunohistochemical detection of 5-hydroxymethyl-cytosine reveals conservation of its tissue distribution between amphibians and mammals

5-Hydroxymethyl-cytosine (5-hmC) is a form of modified cytosine, which has recently attracted a considerable attention due to its potential role in transcriptional regulation. According to several reports 5-hydroxymethyl-cytosine distribution is tissue-specific in mammals. Thus, 5-hmC is enriched in embryonic cell populations and in adult neuronal tissue. Here, we describe a novel method of semi-quantitative immunohistochemical detection of 5-hmC and utilize it to assess the levels of this modification in amphibian tissues. We show that, similar to mammalian embryos, 5-hmC is enriched in axolotl tadpoles compared with adult tissues. Our data demonstrate that 5-hmC distribution is tissue-specific in amphibians, and that strong 5-hmC enrichment in neuronal cells is conserved between amphibians and mammals. In addition, we identify 5-hmC-enriched cell populations that are distributed in amphibian skin and connective tissue in a mosaic manner. Our results illustrate that immunochemistry can be successfully used not only for spatial identification of cells enriched with 5-hmC, but also for the semi-quantitative assessment of the levels of this epigenetic modification in single cells of different tissues. © 2012 Landes Bioscience

5-hydroxymethyl-cytosine enrichment of non-committed cells is not a universal feature of vertebrate development

5-hydroxymethyl-cytosine (5-hmC) is a cytosine modification that is relatively abundant in mammalian pre-implantation embryos and embryonic stem cells (ESC) derived from mammalian blastocysts. Recent observations imply that both 5-hmC and Tet1/2/3 proteins, catalyzing the conversion of 5-methyl-cytosine to 5-hmC, may play an important role in self renewal and differentiation of ESCs. Here we assessed the distribution of 5-hmC in zebrafish and chick embryos and found that, unlike in mammals, 5-hmC is immunochemically undetectable in these systems before the onset of organogenesis. In addition, Tet1/2/3 transcripts are either low or undetectable at corresponding stages of zebrafish development. However, 5-hmC is enriched in later zebrafish and chick embryos and exhibits tissue-specific distribution in adult zebrafish. Our findings show that 5-hmC enrichment of non-committed cells is not a universal feature of vertebrate development and give insights both into evolution of embryonic pluripotency and the potential role of 5-hmC in its regulation. © 2012 Landes Bioscience

Immunostaining for DNA modifications: computational analysis of confocal images

For several decades, 5-methylcytosine (5mC) has been thought to be the only DNA modification with a functional significance in metazoans. The discovery of enzymatic oxidation of 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) as well as detection of N6-methyladenine (6mA) in the DNA of multicellular organisms provided additional degrees of complexity to the epigenetic research. According to a growing body of experimental evidence, these novel DNA modifications may play specific roles in different cellular and developmental processes. Importantly, as some of these marks (e. g. 5hmC, 5fC and 5caC) exhibit tissue- and developmental stage-specific occurrence in vertebrates, immunochemistry represents an important tool allowing assessment of spatial distribution of DNA modifications in different biological contexts. Here the methods for computational analysis of DNA modifications visualized by immunostaining followed by confocal microscopy are described. Specifically, the generation of 2.5 dimension (2.5D) signal intensity plots, signal intensity profiles, quantification of staining intensity in multiple cells and determination of signal colocalization coefficients are shown. Collectively, these techniques may be operational in evaluating the levels and localization of these DNA modifications in the nucleus, contributing to elucidating their biological roles in metazoans

5-hydroxymethyl-cytosine enrichment of non-committed cells is not a universal feature of vertebrate development

5-hydroxymethyl-cytosine (5-hmc) is a cytosine modification that is relatively abundant in mammalian pre-implantation embryos and embryonic stem cells (Esc) derived from mammalian blastocysts. Recent observations imply that both 5-hmc and Tet1/2/3 proteins, catalyzing the conversion of 5-methyl-cytosine to 5-hmc, may play an important role in self renewal and differentiation of Escs. here we assessed the distribution of 5-hmc in zebrafish and chick embryos and found that, unlike in mammals, 5-hmc is immunochemically undetectable in these systems before the onset of organogenesis. In addition, Tet1/2/3 transcripts are either low or undetectable at corresponding stages of zebrafish development. however, 5-hmc is enriched in later zebrafish and chick embryos and exhibits tissue-specific distribution in adult zebrafish. Our findings show that 5-hmc enrichment of non-committed cells is not a universal feature of vertebrate development and give insights both into evolution of embryonic pluripotency and the potential role of 5-hmc in its regulation