Succinate dehydrogenase activity in various parts of the rat heart in systole and diastole

After fixation of the rat heart at the state of systole and diastole, succinate dehydrogenase (SDH) activity has been revealed. In distributions of the muscle fibers according to their optic density, statistic characteristics connected with the first four moments have been taken into consideration. SDH activity is different in five cardiac parts. Individual changeability is more pronounced in the all cardiac parts at diastole. In comparison to the internal one at systole, it is lower in the all parts, and at diastole--in the atria only. The ratio of the internal and individual changeability is in such a state, that it should be taken into account in all statistical calculations, connected with SDH activity determination in cardiomyocytes.     

Ultrastructural and morphometric characteristics of cardiomyocyte mitochondriomes of several invertebrate species. II. Heart ventricle cardiomyocyte mitochondriome of several gastropod mollusks

The mitochondrial ultrastructure in ventricle cardiomyocytes of three gastropod molluscs (Clione limacina, Helix pomatia, Lymnaea stagnalis) has been studied. Mitochondria in cardiomyocytes of these molluscs are connected by intermitochondrial contacts of the same morphology as intermitochondrial contacts in vertebrate cardiomyocytes. Their numbers in cardiomyocytes of the above molluscs being, respectively, 61, 35.1 and 29.2 contacts per 100 mitochondria. In Clione limacina cardiomyocyte contractile elements located on the periphery of cell occupy 21.1% of the cytoplasm volume. Mitochondria form a core making large dense central accumulations taking up 54.9% of the cytoplasm volume. Numerous mitochondria have vesicular or tubular cristae and light matrix. Unlike cardiomyocytes of Clione limacina, in Helix pomatia and Lymnaea stagnalis contractile material predominates in cardiomyocytes occupying 43.7% and 49.2% of the cytoplasm volume, respectively. Mitochondria located on the periphery and in the center of cardiomyocytes in Lymnaea stagnalis and Helix pomatia occupy 31 and 32.5% of the cytoplasma volume, respectively. Mitochondria in cardiomyocytes of both these molluscs have plastic cristae and dense matrix. The differences in cardiomyocyte mitochondriom organization in the studied molluscs can be explained by different functional heart loading in these due to different levels of their locomotor activity.     

Electron microscopic study of the innervation of the heart conduction system in rats]

A systemic quantitative electron microscopic analysis on innervation of the sinus node, the atrioventricular node, the bundle of His and its pedicles within the interventricular septum has been performed in intact hearts of mature rats. The data have been obtained on the size of nonmyelinated and myelinated nerve fibres, efferent and afferent terminals within different parts of the cardiac conductive system, their interconnection with specialized cardiomyocytes have been described. Application of certain methods for electron microscopic investigation on the innervation of mammalian cardiac conductive system has been discussed.     

The effects of cardioactive drugs on cardiomyocytes derived from human induced pluripotent stem cells

Developing effective drug therapies for arrhythmic diseases is hampered by the fact that the same drug can work well in some individuals but not in others. Human induced pluripotent stem (iPS) cells have been vetted as useful tools for drug screening. However, cardioactive drugs have not been shown to have the same effects on iPS cell-derived human cardiomyocytes as on embryonic stem (ES) cell-derived cardiomyocytes or human cardiomyocytes in a clinical setting. Here we show that current cardioactive drugs affect the beating frequency and contractility of iPS cell-derived cardiomyocytes in much the same way as they do ES cell-derived cardiomyocytes, and the results were compatible with empirical results in the clinic. Thus, human iPS cells could become an attractive tool to investigate the effects of cardioactive drugs at the individual level and to screen for individually tailored drugs against cardiac arrhythmic diseases.     

Evidence for the impact of BAG3 on electrophysiological activity of primary culture of neonatal cardiomyocytes

Homeostasis of proteins involved in contractility of individual cardiomyocytes and those coupling adjacent cells is of critical importance as any abnormalities in cardiac electrical conduction may result in cardiac irregular activity and heart failure. Bcl2-associated athanogene 3 (BAG3) is a stress-induced protein whose role in stabilizing myofibril proteins as well as protein quality control pathways, especially in the cardiac tissue, has captured much attention. Mutations of BAG3 have been implicated in the pathogenesis of cardiac complications such as dilated cardiomyopathy. In this study, we have used an in vitro model of neonatal rat ventricular cardiomyocytes to investigate potential impacts of BAG3 on electrophysiological activity by employing the microelectrode array (MEA) technology. Our MEA data showed that BAG3 plays an important role in the cardiac signal generation as reduced levels of BAG3 led to lower signal frequency and amplitude. Our analysis also revealed that BAG3 is essential to the signal propagation throughout the myocardium, as the MEA data-based conduction velocity, connectivity degree, activation time, and synchrony were adversely affected by BAG3 knockdown. Moreover, BAG3 deficiency was demonstrated to be connected with the emergence of independently beating clusters of cardiomyocytes. On the other hand, BAG3 overexpression improved the activity of cardiomyocytes in terms of electrical signal amplitude and connectivity degree. Overall, by providing more in-depth analyses and characterization of electrophysiological parameters, this study reveals that BAG3 is of critical importance for electrical activity of neonatal cardiomyocytes.     

The number and size of human ventricular cardiomyocytes and the number of nuclei in them

The study of isolated cardiomyocytes after alkaline dissociation allowed to reveal an individual variability of the absolute number and average volume of cells, as well as characteristics of these indices in the left and right ventricle myocardia of the dead adult patients' hearts. On the basis of the analysis of these indices and of other parameters of isolated cardiomyocytes, a possibility of hyperplasia of nuclei and of muscle cells themselves during their hypertrophy is discussed.     

Size of cardiomyocytes of the left portion of the heart in adults

As demonstrates the investigation of cardiomyocytes of the left cardiac parts performed in 16 practically healthy mature persons died in accidents, the average area of the cells projections in the auricular atria makes 644 +/- 15 mcm2, and the portion of the multinuclear cells--2.27%. In the ventricular wall and in the papillar muscle these parameters are 831 +/- 16, 700 +/- 30 mcm2 and 11.25, 12.62%, respectively. Distribution of nuclei in the cardiomyocytes of all the cardiac zones studied is characterized by a predominance of tetra-, hypertetra- and octaploid classes, therefore the average amount of DNA in the nuclei reaches certain high values. Differences in size of the cardiomyocytes revealed in various parts of the heart are explained at their unequal loading under pressure.     

Ultrastructure and morphology of the mitochondriome of cardiomyocytes from invertebrates. I. The mitochondriome of cardiomyocytes from insects

The cardiomyocyte mitochondrial ultrastructure of two insect species (the American cockroach Periplaneta americana, and a dragonfly Aeschna sp.) has been studied. Mitochondria in cardiomyocytes of these insects are connected by intermitochondrial contacts, similar in morphology to vertebrate intermitochondrial contacts. The number of intermitochondrial contacts differs in cardiomyocytes of the studied insects, numbering 12 and 18 per 100 mitochondria in cardiomyocytes of the cockroach and dragonfly, respectively, which is due presumably to differences in activity of these insects. Cardiomyocytes of both species have several features in common. It was shown that cross-striated myofibrils oriented in different directions occupy 50-58% of the cytoplasmic volume, while mitochondria cover only 16-18%. The pattern of mitochondrial localization differs in cardiomyocytes of the two studied insects. In the cockroach, cardiomyocyte mitochondria are seen both in the center of the cell and on its periphery, in protrusions; whereas in the dragonfly, mitochondria of cardiomyocytes are confined to the protrusions of the abluminal cell side. Mitochondrial profiles are small, their packing is not dense. Mitochondria in cardiomyocytes of these insects have few plastic cristae and dense matrix.     

Extracellular matrix dependence of organization of the cardiomyocyte contractile apparatus

In the process of cardiomyocyte cultivation, there occurs rearrangement of their contractile apparatus with transformation of typical myofibrils into structures of nonmuscle type and with subsequent restoration of the initial organization. The causes and mechanisms of the rearrangements described are unknown. In this study, it was shown that, at cultivation of cardiomyocytes on the individual proteins of the extracellular matrix (ECM), as well as on the matrix produced either by cardiac fibroblasts or by similar cardiomyocytes, the period in which the contractile apparatus is in a converted state is shortened, whereas soluble factors of the conditioned cultural medium do not produce any obvious effect on the above-indicated rearrangements. Using the method of extracellular matrix isolation adapted for cardiomyocytes, we have confirmed that, in the process of cultivation, cardiomyocytes themselves produce ECM that differs from ECM of cardiac fibroblasts by composition and varies with culturing time.     

吸入麻醉药预处理心肌保护作用的临床研究进展

心肌梗死是围术期最严重的并发症之一,所以减少围术期心肌梗死的风险对于围术期麻醉十分重要。大量实验研究证明,吸入麻醉药预处理可以有效减轻心肌的缺血/再灌注损伤,减少心肌梗死范围,促进心脏功能的恢复。麻醉药预处理是一个复杂的过程,这一过程触发了两个不同的时相。第一,简称为早期预处理(E预处理),包括心肌细胞内有保护作用的酶的激活;第二,称为晚期预处理(L预处理),依赖于新的心肌保护蛋白的从头合成。虽然早期预处理和晚期预处理对心肌细胞的影响是挥发性麻醉药心脏保护作用的关键,但他们对冠状动脉内皮细胞的影响也很重要,这一机制可能改善了冠状动脉手术患者的长期预后。挥发性麻醉药,对改善围术期有心肌梗死风险的非心脏手术患者的预后,尚没有得到有明确意义的证实。     

Stability of beating frequency in cardiac myocytes by their community effect measured by agarose microchamber chip

To understand the contribution of community effect on the stability of beating frequency in cardiac myocyte cell groups, the stepwise network formation of cells as the reconstructive approach using the on-chip agarose microchamber cell microcultivation system with photo-thermal etching method was applied. In the system, the shapes of agarose microstructures were changed step by step with photo-thermal etching of agarose-layer of the chip using a 1064-nm infrared focused laser beam to increase the interaction of cardiac myocyte cells during cultivation. First, individual rat cardiac myocyte in each microstructure were cultivated under isolated condition, and then connected them one by one through newly-created microchannels by photo-thermal etching to compare the contribution of community size for the magnitude of beating stability of the cell groups. Though the isolated individual cells have 50% fluctuation of beating frequency, their stability increased as the number of connected cells increased. And finally when the number reached to eight cells, they stabilized around the 10% fluctuation, which was the same magnitude of the tissue model cultivated on the dish. The result indicates the importance of the community size of cells to stabilize their performance for making cell-network model for using cells for monitoring their functions like the tissue model.     

Morphometric research on the cardiomyocytes in cardiac hyperfunction]

By means of a complex morphometrical method special quantitative parameters of cardiomyocytes have been studied in the cardiac parts of 37 dogs, 6 months after right-sided pulmonectomy, and of 12 control dogs. The cardiac hyperfunction is accompanied with an increased mass of the heart's parts, hypertrophy of the right ventricle and right atrium dominating. Hypertrophy of myocardial parts takes place mainly at the expense of the increasing length and width of cardiomyocytes. This causes disorganization and disorder of morphological systems and an essential decrease in compensatory possibilities of the hyperfunctioning heart parts.     

Individual anatomic variability of particular thalamic nuclei in children and adolescents with respect to stereotaxic coordinates

As demonstrate macro-microscopic and stereologic investigations performed in 120 preparations of children and adolescent brain, the anterior, medial and lateral nuclei in the thalamus have a complex geometric form which does not change with age. Their linear dimentions and stereotaxic coordinates increase somewhat. These changes are described by means of mathematical models as regression equations. Individual changeability of the nuclear groups, situating within the studied ones are evidently subjugated to the same regularities. The individual variants revealed are not connected with side of the brain, form of the skull and sex. The combination of the methods used gives reliable results.     

Quantitative characteristics of the inferior parietal cortical formation of the chimpanzee cerebrum

In histological preparations of the left and right hemispheres a layer-by-layer investigation of cyto- and myeloarchitectonics of the radial system of fibers in the inferior parietal formation (fields 40, 39) has been performed; it is the homologue of the human inferior parietal formation and in chimpanzee it is immediately connected with manipulatory activity. The width of the cytoarchitectonic layers is measured, and in the fibrous layers 4, 5 and 6--also caliber (diameter) of the fibrous fasciculi and fibers in the fasciculi. Statistical parameters of distribution density of the layer widths, caliber of the fasciculi and fibers are estimated. The degree of their changeability is great. The variability coefficient of the layer widths is statistically significant, and there are no interhemispheric differences in width of analogous layers of both fields. Caliber of the fasciculi in the left and right hemispheres is distributed asymmetrically, being greater in the field 39 than in the field 40 of the right hemisphere. Similar regularity in the field 39 is noted in distribution of fibers. Overwhelming majority of fibers in the field 39 have, in comparison with the field 40, greater caliber. The data obtained demonstrate heteromorphism of the radial system in the fibers of the chimpanzee inferior parietal formation. This, evidently, ensures conduction of impulses of various modality.     

A MICRO-DROP APPLICATOR AND ITS USE FOR THE TREATMENT OF CERTAIN SMALL INSECTS WITH LIQUID INSECTICIDE

An instrument has been constructed for applying minute drops of liquid insecticide to selected parts of individual insects. The principle of the instrument, which is not original, is to express minute amounts of liquid from a hypodermic syringe, and blow the drop from the needle tip by a puff of air. This model is considered to have advantages over previous models working on the same principle. In particular, a very convenient form of fitting was developed for blowing the drop from the needle tip, and fittings of this type could be incorporated in existing similar applicators with little modification of the latter. The high directional accuracy of dropfire necessary for treating selected parts of small insects was achieved.
An immersion method was developed for determining the sizes of individual drops of Shell oil P₃₁ delivered, and hence the uncontrolled variability of drop-volume. It showed that the standard deviation in the volumes of drops intended to be equal in size was about 9% of a mean volume of 0.017 μ1. falling progressively to about 2% as the mean volume increased to 0.33 μl. These standard deviations, however, are of limited value, because the sizes of successive drops delivered appeared frequently to be correlated. A method in which the drops are delivered on to an oleophobic surface for measurement of the diameters of the oil lenses formed was found insufficiently accurate for determining the relative variability of drops of P_31.
A suction method was devised for manipulating, without anaesthetization or chilling, small beetles such as Calandra and Tribolium. This enabled large numbers to be individually treated with the applicator in a reasonable time. A similar but slower method was used successfully with the larvae of Ephestia. The biological results achieved have been very satisfactory.
The effects of uncontrolled variation in drop size on dose-response data are discussed.     

Cardiomyocyte ploidy levels in birds with different growth rates

Cytofluorimetric study of ploidy levels in ventricular cardiomyocytes was carried out on 36 adult bird species belonging to 10 orders as well as on the quail Coturnix coturnix, of different ages. It was shown that polyploidization of quail cardiomyocytes occurs during the first 40 days after hatching and ends by the time growth is completed. In adult birds, the cardiomyocyte ploidy hardly changed at all. Interspecies comparison revealed that in the adult bird myocardium 2cx2 myocytes are predominant, accounting for at least 50% of the cell population. Multinuclear cells with three to eight diploid nuclei were widespread. The percentage of such cells was five to six times higher in precocial species than in altricial birds of the same weight. Myocytes with polyploid nuclei were rare. A significant interspecies variability of cardiomyocyte ploidy levels was observed. The most prominent differences were found between the precocial and the altricial birds. The mean number of genomes in cells correlated both with the body mass and with the growth rate of the birds. The differences between the precocial and altricial birds disappeared when a statistical method was used to eliminate the effect of the growth rate, but did not when the effect of body mass was eliminated. Among the altricial birds, which are generally immobile during growth, the cardiomyocyte ploidy levels also correlated more closely with growth rate than with body mass. The opposite was observed in the precocial birds, which are highly mobile from the first minutes of life. We conclude that the interspecies variability of bird cardiomyocyte ploidy levels is a result of changes in the balance between the cardiac functional load and the growth rate; this is manifested at the cellular level as a competition between the proliferation and differentiation of cardiomyocytes. J. Exp. Zool. 289:48-58, 2001.     

Isolation and Culture of Neonatal Mouse Cardiomyocytes

Cultured neonatal cardiomyocytes have long been used to study myofibrillogenesis and myofibrillar functions. Cultured cardiomyocytes allow for easy investigation and manipulation of biochemical pathways, and their effect on the biomechanical properties of spontaneously beating cardiomyocytes.The following 2-day protocol describes the isolation and culture of neonatal mouse cardiomyocytes. We show how to easily dissect hearts from neonates, dissociate the cardiac tissue and enrich cardiomyocytes from the cardiac cell-population. We discuss the usage of different enzyme mixes for cell-dissociation, and their effects on cell-viability. The isolated cardiomyocytes can be subsequently used for a variety of morphological, electrophysiological, biochemical, cell-biological or biomechanical assays. We optimized the protocol for robustness and reproducibility, by using only commercially available solutions and enzyme mixes that show little lot-to-lot variability. We also address common problems associated with the isolation and culture of cardiomyocytes, and offer a variety of options for the optimization of isolation and culture conditions.     

Transformation of adult ventricular myocytes with the temperature sensitive A58 (tsA58) mutant of the SV40 large T antigen

Freshly isolated ventricular myocytes have been used extensively as an adult cardiac model system. Due to their inability to undergo cytokinesisin vitro and their dedifferentiated properties in long-term culture, they can not be used for extended studies. Recent reports tell of the establishment of fetal and neonatal cardiac cell lines and the development of adult cardiomyocytes from transgenic animals. A recent report by Kirshenbaum [1], is the first to demonstrate insertion of genes in to adult ventricular myocytes using viral infection. This paper discusses the infection of primary adult differentiated cardiomyocytes with the SV40 large T antigen and subsequent proliferation under temperature sensitive control. Upon further characterization, the cells could be used as a model to study muscle differentiation and repair as well as adult cardiac cell physiology.