Principal forms of acute damage to cardiomyocytes according to polarization microscopy data

Nepomnyashchikh, L. M.

doi:10.1007/bf02445691

Bull Exp Biol Med

1996

DOI: 10.1007/bf02445691

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Principal forms of acute damage to cardiomyocytes according to polarization microscopy data

L. M. Nepomnyashchikh

Abstract: The principal forms of acute cardiomyocyte damage of metabolic and ischemic genesis are accompanied by changes in the contractile apparatus, which is reflected in double refraction of myofibrils. Thus, polarization microscopy is the most sensitive method allowing one to detect the early stages of cardiomyocyte damage. Individual types of cardiomyocyte lesions are identified on the basis of parallel histological studies and polarization and electron microscopy of prenecrotic alterations and myocardial infarctio… Show more

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1997

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Cited by 7 publications

References 23 publications

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Structural features of acute focal metabolic injuries to somatic muscle fibers caused by dimethylparaphenylenediamine

1999

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The pathological picture of an acute disorder of cell metabolism caused by dimethylparaphenylenediamine consists of typical focal reactions of striated muscles to injury: myofibril contractures and intracellular myocytolysis. Analysis of the pathological process demonstrates the time course of contracture injuries as a succession of overcontracture (first-third degree contractures) and necrobiotic phases (fourth degree contracture), followed by lump degradation and macrophagal resorption. Two polar types of contractures are clearly differentiated by the size, shape, and structural and metabolic characteristics of target fibers: "ribbons " and "medallions". Disappearance of Z strips, disaggregation, disorientation, and fragmentation of myofibrils are typical of intracellular myocytolysis. The observed changes form the morphological basis for acute disease of striated muscles. Key Words: metabolic injuries; somatic muscles; polarization microscopyThe histological picture of pronounced clinical manifestations of somatic muscle disease reflects, as a rule, the interactions between destructive and compensatory-adaptive processes running under heterogeneous structural and metabolic conditions at all levels. The mechanism underlying pathological changes can be elucidated by extrapolation of experimental data characterizing the morphological manifestations during a myopathological process.General mechanisms underlying metabolic injuries to muscle fibers include primary and secondary disorders of energy metabolism, including those caused by blocking of mitochondrial enzyme systems [10,[12][13][14][15]. We analyzed the structural features and evolution of acute focal dystrophic injuries to the skeletal muscles caused by dimethylparaphenylenediamine, a blocker of respiratory enzymes and redox processes in the cell [9]. MATERIALS AND METHODSAcute metabolic injuries to somatic muscle fibers were induced by a single subcutaneous injection of dimethylparaphenylenediamine in a dose of 2.5 mg/100 g [5,9]. Thirty-nine male Wistar rats weighing 150-300 g were used, 14 of then were controls. The animals were sacrificed 6-8 h, 1, 3, and 5 days postinjection.For making the crural muscle preparation in the "strain at rest" state [5], the left hind limb was amputated at the level of the hip joint after decapitation, the skin was rapidly separated, and the whole limb was fixed in cold (4~ 4% paraformaldehyde in Mil-0007-4888/99/0006-0640522.00 9

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Structural features of acute focal metabolic injuries to somatic muscle fibers caused by dimethylparaphenylenediamine

1999

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Photochemical fluorochrome staining for detection of prenecrotic changes in cardiomyocyte myofibrils

Nepomnyashchikh

Tsimmerman

1999

Bull Exp Biol Med

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Ischemic and metabolic damages to cardiomyocytes induce immediate ultrastructural changes in their myofibrils. Photochemical fluorochromize staining of prenecrotic cardiomyocytes allows to determine with high sensitivity local redistribution of sarcomere mass. It is shown that different types of destructive processes in cardiomyocyte myofibrils underlie similar polarization microscopic picture.Key Words: necrobiosis; cardiomyocytes; myofibrils; luminescence and polarization microscopy Double refraction of myosin fibrils of sarcomeres allowed to use polarization microscopy for detection of the most early reactions of cardiomyocytes (CMC) to damage [3][4][5][6]10,11,15]. These results provide the basis for classification of CMC alterations. We proposed a method of photochemical fluorochromize staining for detection of early necrobiotic changes in CMC based on the formation of luminescent products in CMC after short wave UV (SUV) irradiation [12][13][14].Polarized light microscopy of CMC is based on changes in anisotropy and topography of myosin filaments in altered cell. Here we used photochemical fluorochromize staining in polarization microscopy of myofibrils during CMC alterations. MATERIALS AND METHODSOcclusion myocardial infarction was modeled in 39 male C57BI mice weighing 17-20 g. The left carotid artery was ligated at the level of the middle third of the left ventricle under Nembutal narcosis (0,5%, 1.25Laboratory of General Pathological Anatomy, Institute of Regional Pathology and Pathomorphologyl Institute of Cytology and Genetics, Siberian Division of the Russian Academy of Medical Sciences, Novosibirsk ml/100 g body weight). The animals were sacrificed 5 min and 1 day after coronary occlusion. Catecholamine-induced damage to the myocardium was modeled in 34 male Wistar rats weighing 160-270 g. Isopropylnoradrenaline (8-10 mg/100 g) was injected intraperitoneally 0.5-18 h before decapitation.After decapitation the heart was isolated and incubated at 0~ to termination of contraction and muscle relaxation. The atria were separated and the heart was cut along through the ventricles and septum. The tissue was washed in a cold neutral buffer. Some samples were frozen in liquid nitrogen and 10-g sections were prepared on an MK-25 cryostate at -18~ mounted on warm glass slides, dried on air, and exposed to phosphorus pentoxide. Fixed and paraffin-embedded tissue samples were cut into 7-~t sections. Before fluorochromize staining the sections were treated with xylene, alcohol, ammonia, and water to ensure complete removal of paraffin and formaldehyde [2].Luminescent, polarization, interference, and brightfield microscopy was used in the study. The preparations were stained by the following techniques: hematoxylin and eosin staining combined with Perls reaction, PAS-reaction combined with colloid iron, according to Hale, and hematoxylin orange, PAS-reaction after amylase treatment and hematoxylin-picrofuchsin 0007-4888/99/0009-0967522.00 9Kluwer Academic/Plenum Publishers

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Electron microscopic characteristic of major types of acute damage to cardiomyocytes

Nepomnyashchikh

1997

Bull Exp Biol Med

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The major (independent) types of acute damage to cardiomyocytes: myofibrillar contractures, intracellular myocytolysis, primary crumpy degradation, and cytolysis are characterized ultrastructurally. It is shown that dystrophic and necrobiotic changes in the myocardium are mosaic, implying that prior to electron microscopic examination cardiomyocytes should be studied in polarized light. Key Words: acute damage to cardiomyocytes; myofibrillar contracture; intracellular myoeytolysis; primary clumpy degradation of myofibrils; eytolysis; electron microscopyInvestigation of patho-and morphogenesis of the damage to cardiomyocytes is crucial for cardiological diagnosis and prognosis. Pathomorphological diagnosis of acute myocardial damage is based on the presence of necrotic, inflammatory, and sclerotic alterations [9,11,15] which reflect the outcome of the damage to cardiomyocytes (CM). With the aid of histochemical enzyme and luminescence techniques necrobiotic processes can be identified in autopsy material 3-4 h after their beginning [1]. A great progress in the diagnosis of acute damage to the myocardium has been achieved with the use of polarizing microscopy. This method allows one not only to identify the early damage to CM, but also to classify individual types of this damage that lay the basement for alterative myocardial insufficiency [2,3,6,8]. More insight into structural mechanisms underlying acute myocardial damage can be provided by electron microscopy. It should be noted, however, that damaged CM are often studied without considering the data obtained by polarizing and electron microscopy. Our aim was to examine damaged CM under an electron microscope after identification in polarized light. MATERIALS AND METHODSAcute myocardial damage was produced with cardiotoxic doses of epinephrine and isopropylnorepinephrine, cobalt chloride, functional overload, and ventricular fibrillations induced by electrical current [11. Myocardial infarction was provoked by coronary occlusion [2]. The myocardium was investigated 5 min--48 h after application of damaging factor. Myocardium of people died from acute cardiac failure, acute coronary insufficiency, myocardial infarction, and noncardiological diseases was also studied [1,21. Specimens (left papillary muscle) for electron microscopy [7] were fixed in 4% paraformaldehyde for 2 h at 4~ postfixed in 1-2% OsO 4 solution for 2 h, and embedded in Epon-Araldite. When necessary, the same specimens were embedded in styrene-butyl methacrylate, since semithin styrenemethacrylate sections can be more efficiently investigated in polarized light.

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Principal forms of acute damage to cardiomyocytes according to polarization microscopy data

Cited by 7 publications

References 23 publications

Structural features of acute focal metabolic injuries to somatic muscle fibers caused by dimethylparaphenylenediamine

Structural features of acute focal metabolic injuries to somatic muscle fibers caused by dimethylparaphenylenediamine

Photochemical fluorochrome staining for detection of prenecrotic changes in cardiomyocyte myofibrils

Electron microscopic characteristic of major types of acute damage to cardiomyocytes

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