Assay of acetylcholinesterase activity and elemental composition in brain compartments by electron probe microanalysis

Pogorelov, A. G.; Budantsev, A. Yu.; Pogorelova, V. N.; Mizin, I.

doi:10.1016/s1385-299x(96)00012-8

Cited by 6 publications

(7 citation statements)

References 12 publications

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“…Due to design of these devices and stability of physical efforts, the cutting step (critical parameter for estimating element concentrations in EPMA) is highly accurate [5,53,55,57]. In order to rule out the impact of the cryosection thickness variations, a special method was developed, when the results of each measurement were standardized with consideration for the intensity of inhibitory X-ray radiation proportional to the local bulky thickness of the section [40,41].…”

Section: Group Sodium Potassiummentioning

confidence: 99%

“…The adequacy of the method of "embedded" sections for EPMA was verified on tissues [55,57] and in comparative experiments on preparations of primary cardiomyocyte culture [8,56]. The method was used with lyophilized cryosections of primary culture of cardiomyocytes; data for isolated myocyte were reported [68].…”

Section: Group Sodium Potassiummentioning

confidence: 99%

“…This problem is essential for cells with different in vitro and in vivo status: hepatocyte [69], cardiomyocyte [8,68], and neuron [57]. In this respect it seems interesting to compare the element balance in a mammalian embryonic totipotent cell at the morula or blastocyst stage as analog of cell clone and in a clonogenic structure grown from a stem cell, e. g., human neurosphere.…”

Section: Group Sodium Potassiummentioning

confidence: 99%

“…Due to succession of events, no aqueous phase forms and no solvents are needed in any of the variants. Cryomethods serve not only as instruments for preservation of a life-time distribution of element composition, but promote optimal sensitivity and spatial resolution of EPMA [12,53,55,57].…”

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confidence: 99%

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Electron probe microanalysis of cytoplasmic concentrations of elements in a single cell in culture and suspension

Pogorelov

Goldstein

2006

Bull Exp Biol Med

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Electron probe microanalysis is a method for evaluation of concentrations of elements at the subcellular level, effective in studies of an individual cell in culture or suspension. Intracellular concentrations of cytoplasmic ions (K+, Na+, Cl-) most rapidly and markedly reacting to changes is an adequate criterion for evaluating the traumatism of manipulations used in cell technologies. Using electron probe microanalysis it will be possible to develop special procedures (for example, therapeutic cloning) for reprogramming or cryopreservation, when intactness of intracellular balance of element serves as the criterion of cell status.

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Section: Group Sodium Potassiummentioning

confidence: 99%

Section: Group Sodium Potassiummentioning

confidence: 99%

Section: Group Sodium Potassiummentioning

confidence: 99%

mentioning

confidence: 99%

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Electron probe microanalysis of cytoplasmic concentrations of elements in a single cell in culture and suspension

Pogorelov

Goldstein

2006

Bull Exp Biol Med

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“…The technique of embryo section preparation and the parameters of electron probe microanalysis were discussed earlier [8,9].…”

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confidence: 99%

Analysis of the diffusion and accumulation of potassium in mouse zygotes mediated by the cyclic activity of 240 pS K+ channel

et al. 2005

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Change in K + concentration induces key molecular genetic transformations: gene expression [1], DNA recombination/degradation [2], and (de)polymerization of actin [3]. The oscillation of the intracellular potassium level of early mammalian embryos was predicted from the cyclicity of the 240 pS K + channel whose activity is synchronized with the phases of the cell cycle [4][5]. Therefore, the purposes of this work were the following: (1) to study the potassium content and distribution in the cytoplasm of the mouse zygote at different stages of its development and (2) on the basis of the experimental data, elaborate the mathematical model of K + diffusion during the first cell cycle. The obtained results allowed us to put forward the hypothesis that the mechanism of the approach of pronuclei during the G 1 /S transition is mediated by the polymerization of actin-like proteins in the cell.It is known that G-actin exposed to physiologically high K + concentration is polymerized into the F form after binding the cation [6,7]. The storage of potassium, if it is present, can significantly change the electrolyte diffusion rate in the cytoplasm. This rate is suggested to be comparable to the rate in a water solution. This also indicates the appearance of a gradient of the element concentration between the periphery and the central region of the mouse zygote. The K + concentration should be changed with time according to the activity of the 240 pS K + channel. The electron probe microanalysis of potassium compartmentalization in the embryo cytoplasm was employed to test our hypothesis.This method is based on recording the intensity of the K α line of X-radiation excited in the zygote structure by accelerated electrons of the probe, which is formed in the electron microscope. The technique of embryo section preparation and the parameters of electron probe microanalysis were discussed earlier [8,9].The monitoring of embryo morphology and measurement of cytoplasmic potassium concentration in it was performed in 2-µ m slices using a JSM U3 electron microscope (JEOL, Japan). The results of the quantitative electron probe analysis are shown in the table.Analysis of potassium distribution in the cytoplasm of mouse zygote. After the fertilization, the potassium concentration changes during the zygote development (table). It is noteworthy that a significant difference between peripheral cytoplasm of embryo zygote and its central part was observed during the transition between G 1 /S and G 2 /M phases. These data suggest that the parameters of K + diffusion in the internal medium of the zygote do not correspond to those in a water solution.The analysis of the spatiotemporal characteristics of the distribution of cytoplasmic potassium in the zygote during different phases of its development (table) allows us to formulate the boundary and initial conditions for the calculation of the diffusion coefficient. At the initial moment, potassium concentration in the cytoplasm is distributed uniformly and is similar to that in the oocyte duri...

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The Role of δ-Opioid Receptors in Brain Ionic Homeostasis Under Physiological Condition

Chao¹,

Xia²

2015

Neural Functions of the Delta-Opioid Receptor

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Assay of acetylcholinesterase activity and elemental composition in brain compartments by electron probe microanalysis

Cited by 6 publications

References 12 publications

Electron probe microanalysis of cytoplasmic concentrations of elements in a single cell in culture and suspension

Electron probe microanalysis of cytoplasmic concentrations of elements in a single cell in culture and suspension

Analysis of the diffusion and accumulation of potassium in mouse zygotes mediated by the cyclic activity of 240 pS K+ channel

The Role of δ-Opioid Receptors in Brain Ionic Homeostasis Under Physiological Condition

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