Cellular and subcellular localization of catalase in the heart of transgenic mice.

Previous studies have described a cardiac-specific, catalase-overexpressing transgenic mouse model that was used to study myocardial oxidative injury. This study was undertaken to demonstrate cellular and subcellular localization of catalase in the hearts of transgenic mice. By the light microscopic immunoperoxidase method, we found that the overexpressed catalase was exclusively localized in cardiomyocytes. The ratios of immunoreactive cardiomyocytes in the heart were quite different among three transgenic lines examined but agreed with the elevated levels of catalase activity. In the cardiac blood vessels, positive cells were found in the walls of pulmonary veins and the vena cava, which consist of cardiomyocytes, but not in the pulmonary arteries, aorta, or cardiac valves. The electron microscopic immunogold method revealed that the elevated catalase was in sarcoplasm, nucleus, and peroxisomes, but not in mitochondria. In contrast to these distributions, catalase in the non-transgenic cardiomyocytes was in peroxisomes only. In addition, the number and size of peroxisomes in the transgenic cardiomyocytes were markedly increased, but no other ultrastructural changes were observed in comparison with those of non-transgenic mice. These results demonstrated that the elevated catalase in transgenic mouse heart is localized in cardiomyocytes and is distributed to peroxisomal and extraperoxisomal, but not mitochondrial, compartments.     

Acrolein,an Environmental Toxin,Induces Cardiomyocyte Apoptosis via Elevated Intracellular Calcium and Free Radicals

Acrolein, an unsaturated aldehyde, is an environmental toxin known to inhibit mitochondrial electron transport chain in brain and induce lipid peroxidation and apoptosis. However, the nature of the effects of acrolein on cardiac function and myocardium is not known. The objective of this study is to examine whether acrolein induces apoptosis in cardiomyocytes and alters cytosolic calcium concentration and the intracellular oxygen free-radical levels. Adult mouse cardiomyocytes exposed to 1 μmol/l of acrolein showed a marked increase in the intracellular oxygen free-radicals and calcium concentration, by 12- and 2-fold, respectively, compared to the resting value. Moreover, the cardiomyocyte viability decreased significantly in a dose-dependent manner by treatment with 25, 50, and 100 μmol/l of acrolein compared to controls. Morphological changes and DNA laddering typical of apoptosis were found in acrolein-exposed cardiomyocytes. Our finding suggested that acrolein caused apoptotic death of adult mice cardiomyocytes by increasing intracellular oxygen free-radicals and calcium concentration.     

MZ15, a monoclonal antibody recognizing keratan sulphate, stains chick tendon

Summary Biochemical, morphometric and electron histochemical methods have failed to demonstrate the presence in tendon of keratan sulphate, a common component of connective tissue proteoglycans. Using a monoclonal antibody specific for keratan sulphate, a positive localization of this molecule was made in the gastrocnemius tendon of stage 44 chicken embryos both at the light and electron microscopical levels.     

Morphological characteristics of the thymus gland of albino rats after prenatal administration of indomethacin

By means of histological, histochemical and electron microscopical methods the thymus of white rat fetuses and offspring has been investigated during various age periods after indomethacin++ influence (2.5 mg/kg). In the fetuses retardation in separation of the gland parenchyma into lobules has been revealed. During the first two weeks of life the section area of the medulla decreases. Amount of lymphoid cells decreases; small and degenerating lymphocytes decrease in their number, while the part of lymphoblasts, middle lymphocytes and figures of mitosis increases. Enzymatic activity in the nervous structures of the organ is inhibited. Some essential disturbances of the intracellular structures are revealed; they demonstrate certain destructive changes. The data obtained show a decreasing function of the thymus after the prenatal influence of indomethacin++ during the first month of life, which is especially manifested during the first two weeks.     

Differential mitochondrial calcium responses in different cell types detected with a mitochondrial calcium fluorescent indicator, mito-GCaMP2

Mitochondrial calcium plays a crucial role in mitochondrial metabolism, cell calcium handling, and cell death. However, some mechanisms concerning mitochondrial calcium regulation are still unknown, especially how mitochondrial calcium couples with cytosolic calcium. In this work, we constructed a novel mitochondrial calcium fluorescent indicator (mito-GCaMP2) by genetic manipulation. Mito-GCaMP2 was imported into mitochondria with high efficiency and the fluorescent signals co-localized with that of tetramethyl rhodamine methyl ester, a mitochondrial membrane potential indicator. The mitochondrial inhibitors specifically decreased the signals of mito-GCaMP2. The apparent K(d) of mito-GCaMP2 was 195.0 nmol/L at pH 8.0 in adult rat cardiomyocytes. Furthermore, we observed that mito-GCaMP2 preferred the alkaline pH surrounding of mitochondria. In HeLa cells, we found that mitochondrial calcium ([Ca(2+)](mito)) responded to the changes of cytosolic calcium ([Ca(2+)](cyto)) induced by histamine or thapasigargin. Moreover, external Ca(2+) (100 μmol/L) directly induced an increase of [Ca(2+)](mito) in permeabilized HeLa cells. However, in rat cardiomyocytes [Ca(2+)](mito) did not respond to cytosolic calcium transients stimulated by electric pacing or caffeine. In permeabilized cardiomyocytes, 600 nmol/L free Ca(2+) repeatedly increased the fluorescent signals of mito-GCaMP2, which excluded the possibility that mito-GCaMP2 lost its function in cardiomyocytes mitochondria. These results showed that the response of mitochondrial calcium is diverse in different cell lineages and suggested that mitochondria in cardiomyocytes may have a special defense mechanism to control calcium flux.     

Histochemistry and cytochemistry of glucose-6-phosphate dehydrogenase

Histochemistry and cytochemistry of glucose-6-phosphate dehydrogenase has found many applications in biomedical research. However, up to several years ago, the methods used often appeared to be unreliable because many artefacts occurred during processing and staining of tissue sections or cells. The development of histochemical methods preventing loss or redistribution of the enzyme by using either polyvinyl alcohol as a stabilizer or a semipermeable membrane interposed between tissue section and incubation medium, has lead to progress in the topochemical localization of glucose-6-phosphate dehydrogenase. Optimization of incubation conditions has further increased the precision of histochemical methods. Precise cytochemical methods have been developed either by the use of a polyacrylamide carrier in which individual cells have been incorporated before staining or by including polyvinyl alcohol in the incubation medium. In the present text, these methods for the histochemical and cytochemical localization of glucose-6-phosphate dehydrogenase for light microscopical and electron microscopical purposes are extensively discussed along with immunocytochemical techniques. Moreover, the validity of the staining methods is considered both for the localization of glucose-6-phosphate dehydrogenase activity in cells and tissues and for cytophotometric analysis. Finally, many applications of the methods are reviewed in the fields of functional heterogeneity of tissues, early diagnosis of carcinoma, effects of xenobiotics on cellular metabolism, diagnosis of inherited glucose-6-phosphate dehydrogenase deficiency, analysis of steroid-production in reproductive organs, and quality control of oocytes of mammals. It is concluded that the use of histochemistry and cytochemistry of glucose-6-phosphate dehydrogenase is of highly significant value in the study of diseased tissues. In many cases, the first pathological change is an increase in glucose-6-phosphate dehydrogenase activity and detection of these early changes in a few cells by histochemical means only, enables prediction of other subsequent abnormal metabolic events. Analysis of glucose-6-phosphate dehydrogenase deficiency in erythrocytes has been improved as well by the development of cytochemical tools. Heterozygous deficiency can now be detected in a reliable way. Cell biological studies of development or maturation of various tissues or cells have profited from the use of histochemistry and cytochemistry of glucose-6-phosphate dehydrogenase activity.(ABSTRACT TRUNCATED AT 400 WORDS)     

The value of the fluorescence histochemistry of biogenic amines in neurotoxicology

Conclusions Acid- and aldehyde-induced fluorescence offers a highly sensitive and specific instrument for the histochemical demonstration of biogenic amines. This technique can be used to advantage for the selective identification of those neuronal structures that contain biogenic amines, namely the peripheral postganglionic sympathetic neurones and the central aminergic neuronal systems.Structural changes of impaired aminergic neurones can be ascertained from their fluorescence microscope image and correlated with light and electron microscopical observations, so that selective neurotoxic changes, such as sympathetic denervation of organs, can be detected and the reversibility of these changes tested.The degree of functional and structural changes occurring in the above neuronal systems can be easily quantified by means of microfluorimetry.The histochemical approach is restricted by the necessity of using fresh and specially fixed tissues. The possibility of numerous pitfalls in the interpretation of histochemical reactions requires the simultaneous use of other optical methods, such as light and electron microscopy, or the testing of the uptake of exogeneous amines or their precursors, whenever the occurrence of neurotoxic effects is to be assessed.     

A light and electron microscope study on oogenesis in the freshwater pulmonate snail Biomphalaria glabrata

In the freshwater snail Biomphalaria glabrata the formation and composition of yolk granules and the role of the follicle cells were studied by histochemical and electron microscopical techniques. The rough endoplasmic reticulum and the Golgi apparatus appeared to be involved in yolk formation, which is a continuous process throughout oogenesis. From the very beginning of yolk formation two main types of yolk granules were distinguished morphologically. However, with histochemical and enzyme cytochemical methods no differences were observed between these types. The granules acquire lysosomal enzymes after oviposition, indicating that their main function is probably digestion of perivitelline fluid, which contains nutrients for the developing embryo.Yolk formation and the activity of the follicle cells were studied in successive stages of oogenesis by quantitative electron microscopy. The data strongly suggest that the follicle cells are involved in the formation of the follicular cavity and hence in the ovulation process.     

不同盐胁迫下小麦叶片渗透性调节和叶绿素荧光特性

盐胁迫对植物伤害机理受到普遍关注。本试验以‘西旱3号’小麦幼苗为材料,通过比较钠盐(150 mmol·L^-1)、钙盐(5、30 mmol·L^-1)单独及其复合胁迫对叶片渗透调节和光合特性的影响,揭示不同盐胁迫对小麦的伤害机理。结果表明: 钠盐或钙盐单独胁迫显著抑制了小麦幼苗根、茎的生长,使叶片可溶性糖和脯氨酸含量、调节性能量耗散电子产量、非光化学猝灭及玉米黄质相对含量均显著增加,而叶绿素a和叶绿素b含量、最大光化学效率、PSⅡ实际光化学效率、光化学猝灭及光合电子传递效率均显著下降。此外,钙盐对小麦幼苗生长的抑制作用更强,钠盐处理下叶片叶绿素含量减少和叶绿素荧光参数降低更显著。除了可溶性蛋白、叶黄素和玉米黄质相对含量以外,低浓度钙盐有效缓解了钠盐诱导其他各指标的变化,而高浓度钙盐进一步增大了钠盐处理小麦幼苗各参数的变化幅度。总之,钠盐和钙盐显著抑制了小麦幼苗的生长,低浓度钙盐能有效缓解钠盐对小麦幼苗的伤害,而高浓度钙盐加剧了钠盐的毒害作用。这均与叶片光合色素含量、光能捕获及光合电子传递的改变有关。此外,渗透调节物质在增强钠盐或钙盐环境中小麦幼苗的抗性方面发挥着重要作用。     

Mitochondrial Adaptations to NaCl. Complex I Is Protected by Anti-Oxidants and Small Heat Shock Proteins, Whereas Complex II Is Protected by Proline and Betaine

High soil sodium (Na) is a common stress in natural and agricultural systems. Roots are usually the first tissues exposed to Na stress and Na stress-related impairment of mitochondrial function is likely to be particularly important in roots. However, neither the effects of NaCl on mitochondrial function, nor its protection by several potential adaptive mechanisms, have been well studied. This study investigated the effects of NaCl stress on maize (Zea mays) mitochondrial electron transport and its relative protection by osmoprotectants (proline, betaine, and sucrose), antioxidants (ascorbate, glutathione, and alpha-tocopherol), antioxidant enzymes (catalase and Cu/Zn-superoxide dismutase), and mitochondrial small heat shock proteins (sHsps). We demonstrate that Complex I electron transport is protected by antioxidants and sHsps, but not osmoprotectants, whereas Complex II is protected only by low concentrations of proline and betaine. These results indicate that NaCl stress damaged Complex I via oxidative stress and suggests that sHsps may protect Complex I as antioxidants, but NaCl damaged Complex II directly. This is the first study to demonstrate that NaCl stress differentially affects Complex I and II in plants and that protection of Complex I and II during NaCl stress is achieved by different mechanisms.     

Lipids at the stages of early osteogenesis in human long tubular bones

In 240 long tubular bone anlagen (LTBA) of extremities in 40 human embryos and prefetuses at the age of 6-12 weeks of the prenatal development the investigation has been performed concerning localization, dynamics of contents and spectrum of neutral lipids and phospholipids. Lipids are stained with Sudan III and IV, phospholipids-with Sudan black with corresponding control extractive methods and quantitative estimation of these substances values. The material for electron microscopical investigation of the lipids is treated according to the method, that preserves their safety. The spectrum of lipids and phospholipids in the LTBA is studied by means of the thin layered chromatography method. The data of the histochemical investigation and chromatography demonstrate decreasing contents of neutral lipids with the gradient from the zone of the preserved cartilage up to the ossification zone of the epiphyseal cartilage of the LTBA in the human embryos and prefetuses. Increased concentration of phospholipids and complication of their spectrum is noted in the areas of intensive deposits of calcium salts. An essential role of the substance of lipid origin is supposed in ossification and mineralization of the human LTBA.     

Protection of Rat Myocardium by Coenzyme Q during Oxidative Stress Induced by Hydrogen Peroxide

Ubiquinone Q(10) (coenzyme Q) is an important component of the mitochondrial electron transport chain and an antioxidant. The purpose of this work was to find out whether an increase in the level of coenzyme Q in the heart changes its maximal working capacity and resistance to oxidative stress. Male Wistar rats were treated with coenzyme Q (10 mg/kg body weight per day) for six weeks, and this increased its content in the myocardium by 63%. The myocardial content of malonic dialdehyde and activities of key antioxidant enzymes were unchanged, except nearly 2.5-fold decrease in the activity of superoxide dismutase. The maximal working capacity of the isolated isovolumic heart did not change, but under conditions of oxidative stress induced by 45-min infusion of hydrogen peroxide (70 micro M) into coronary vessels the contractile function of these hearts decreased significantly more slowly. This was associated with less pronounced lesions in the ultrastructure of cardiomyocytes and lesser disorders in the oxidative metabolism of mitochondria that suggested increased antioxidant protection of the myocardium.     

Expression and characterization of Edg-1 receptors in rat cardiomyocytes: calcium deregulation in response to sphingosine 1-phosphate.

Recent evidence indicates that sphingolipids are produced by the heart during hypoxic stress and by blood platelets during thrombus formation. It is therefore possible that sphingolipids may influence heart cell function by interacting with G-protein-coupled receptors of the Edg family. In the present study, it was found that sphingosine 1-phosphate (Sph1P), the prototypical ligand for Edg receptors, produced calcium overload in rat cardiomyocytes. The cDNA for Edg-1 was cloned from rat cardiomyocytes and, when transfected in an antisense orientation, effectively blocked Edg-1 protein expression and reduced the Sph1P-mediated calcium deregulation. Taken together, these results demonstrate that cardiomyocytes express an extracellular lipid-sensitive receptorsystem that can respond to sphingolipid mediators. Because the major source of Sph1P is from blood platelets, we speculate that Edg-mediated Sph1P negative inotropic and cardiotoxic effects may play important roles in acute myocardial ischemia where Sph1P levels are probably elevated in response to thrombus.     

Calcium microdomains and oxidative stress

The phenomenon of calcium microdomains is firmly established in the field of subcellular physiology. These regions of localized, transient calcium increase are exemplified by the spontaneous 'sparks' released through the ryanodine receptor in myocytes, but include subplasmalemmal microdomains, focal calcium oscillations and microdomains enclosed within organelles, such as the endoplasmic reticulum, golgi and mitochondria. Increasing evidence suggests that oxidative stress regulates both the formation and disappearance of microdomains. Calcium release channels and transporters are all modulated by redox state, while several mechanisms that generate oxidative or nitrosative stress are regulated by calcium. Here, we discuss the evidence for the regulation of calcium microdomains by redox state, and, by way of example, demonstrate that the frequency of calcium sparks in cardiomyocytes is increased in response to oxidative stress. We consider the evidence for the existence of analogous microdomains of reactive oxygen and nitrogen species and suggest that the refinement of imaging techniques for these species might lead to similar concepts. The interaction between Ca(2+) microdomains and proteins that modulate their formation results in a complex and dynamic, spatial signaling mechanism, which is likely to be broadly applicable to different cell types, adding new dimensions to the calcium signaling 'toolkit'.     

Ultrahistochemical localization of Na+-K+ ATPase, Ca2+-ATPase and alkaline phosphatase activity in a calcium-transporting epithelium of a crustacean during moulting

Periodical changes in Na+-K+-ATPase, Ca2+-ATPase and non-specific alkaline-phosphatase activity were observed using cytochemical techniques in the posterior caeca of the crustacean amphipod, Orchestia cavimana, during the moult cycle. These changes were considered in relation to the calcium transport mechanisms in the posterior caecal epithelium. For both ATPases as well as alkaline phosphatase, the specific reaction products were most intense during the pre-exuvial period, i.e. when calcium is slowly transported against a concentration gradient: the localization of Na+-K+-ATPase activity in microvilli and the upper extracellular channels strongly supports the hypothesis that this enzyme is involved in an indirect, sodium-dependent mechanism for the transport of calcium. The detection of Ca2+-ATPase activity in microvilli would seem to indicate that this enzyme plays a role in the direct, active extrusion of Ca2+ at this level. Although the role of alkaline phosphatase in the transport of calcium remains unclear, the histochemical detection of this enzymatic activity throughout the apical part of the caecal epithelium suggests that this enzyme may be involved in calcium secretion. In post-exuvial period, we found only weak specific reaction products, thus indicating a reduced active calcium transport as these ions are rapidly reabsorbed down the concentration gradient.     

Enhanced Connexin-43 and α-Sarcomeric Actin Expression in Cultured Heart Myocytes Exposed to Triiodo-l-thyronine

This study examined whether triiodo-L-thyronine (T3) affects the expression of the major intercellular channel protein, connexin-43, and contractile protein alpha-sarcomeric actin. Cultured cardiomyocytes from newborn rats were treated on day three in culture with 10 or 100 nM T3 and examined 48 and 72 h thereafter. Treated and untreated cells were examined by immunofluorescence and electron microscopy. Expression levels of Cx43 and sarcomeric alpha-actin were monitored by Western blot analysis. Immunofluorescence labeling showed cell membrane location of Cx43 in punctuate gap junctions, whereby fluorescence signal area was significantly higher in cultured cardiomyocytes exposed to T3. This correlated with electron microscopical findings showing increased numbers and size of gap junction profiles, as well as with a significant dose-dependent increase of Cx43 expression detected by Western blot. Immunofluorescence of sarcomeric a-actin was enhanced and its expression increased dose- and time-dependently in T3-treated cultured heart myocytes. However, exposure to the higher dosage (100 nM) of T3 caused mild disintegration of sarcomeric a-actin in some myocytes, suggesting an over-dosage. The results indicate that T3 up-regulates Cx43 and accelerates gap junction formation in cultured neonatal cardiomyocytes. They suggest that thyroid status cannot only modulate the mechanical function of cardiomyocytes but also cell-to-cell communication essential for myocardial electrical and metabolic synchronizations.     

Thermal Stress Promotes Host Mitochondrial Degradation in Symbiotic Cnidarians: Are the Batteries of the Reef Going to Run Out?

The symbiotic relationship between cnidarians and their dinoflagellate symbionts, Symbiodinium spp, which underpins the formation of tropical coral reefs, can be destabilized by rapid changes to environmental conditions. Although some studies have concluded that a breakdown in the symbiosis begins with increased reactive oxygen species (ROS) generation within the symbiont due to a decoupling of photosynthesis, others have reported the release of viable symbionts via a variety of host cell derived mechanisms. We explored an alternative model focused upon changes in host cnidarian mitochondrial integrity in response to thermal stress. Mitochondria are often likened to being batteries of the cell, providing energy in the form of ATP, and controlling cellular pathway activation and ROS generation. The overall morphology of host mitochondria was compared to that of associated symbionts under an experimental thermal stress using confocal and electron microscopy. The results demonstrate that hyperthermic stress induces the degradation of cnidarian host mitochondria that is independent of symbiont cellular deterioration. The potential sites of host mitochondrial disruption were also assessed by measuring changes in the expression of genes associated with electron transport and ATP synthesis using quantitative RT-PCR. The primary site of degradation appeared to be downstream of complex III of the electron transport chain with a significant reduction in host cytochrome c and ATP synthase expression. The consequences of reduced expression could limit the capacity of the host to mitigate ROS generation and maintain both organelle integrity and cellular energy supplies. The disruption of host mitochondria, cellular homeostasis, and subsequent cell death irrespective of symbiont integrity highlights the importance of the host response to thermal stress and in symbiosis dysfunction that has substantial implications for understanding how coral reefs will survive in the face of climate change.     

Osmotic stress alters chromatin condensation and nucleocytoplasmic transport

Osmotic stress is a potent regulator of biological function in many cell types, but its mechanism of action is only partially understood. In this study, we examined whether changes in extracellular osmolality can alter chromatin condensation and the rate of nucleocytoplasmic transport, as potential mechanisms by which osmotic stress can act. Transport of 10 kDa dextran was measured both within and between the nucleus and the cytoplasm using two different photobleaching methods. A mathematical model was developed to describe fluorescence recovery via nucleocytoplasmic transport. As osmolality increased, the diffusion coefficient of dextran decreased in the cytoplasm, but not the nucleus. Hyper-osmotic stress decreased nuclear size and increased nuclear lacunarity, indicating that while the nucleus was getting smaller, the pores and channels interdigitating the chromatin had expanded. The rate of nucleocytoplasmic transport was increased under hyper-osmotic stress but was insensitive to hypo-osmotic stress, consistent with the nonlinear osmotic properties of the nucleus. The mechanism of this osmotic sensitivity appears to be a change in the size and geometry of the nucleus, resulting in a shorter effective diffusion distance for the nucleus. These results may explain physical mechanisms by which osmotic stress can influence intracellular signaling pathways that rely on nucleocytoplasmic transport.     

Ultrahistochemical localization of Na+−K+ ATPase,Ca2+-ATPase and alkaline phosphatase activity in a calcium-transporting epithelium of a crustacean during moulting

Summary Periodical changes in Na+–K+-ATPase, Ca2+–ATPase and non-specific alkaline-phosphatase activity were observed using cytochemical techniques in the posterior caeca of the crustacean amphipod, Orchestia cavimana, during the moult cycle. These changes were considered in relation to the calcium transport mechanisms in the posterior caecal epithelium. For both ATPases as well as alkaline phosphatase, the specific reaction products were most intense during the pre-exuvial period, i.e. when calcium is slowly transported against a concentration gradient: the localization of Na+–K+-ATPase activity in microvilli and the upper extracellular channels strongly supports the hypothesis that this enzyme is involved in an indirect, sodium-dependent mechanism for the transport of calcium. The detection of Ca2+-ATPase activity in microvilli would seem to indicate that this enzyme plays a role in the direct, active extrusion of Ca2+ at this level. Although the role of alkaline phosphatase in the transport of calcium remains unclear, the histochemical detection of this enzymatic activity throughout the apical part of the caecal epithelium suggests that this enzyme may be involved in calcium secretion. In post-exuvial period, we found only weak specific reaction products, thus indicating a reduced active calcium transport as these ions are rapidly reabsorbed down the concentration gradient.     

Inhibition of ErbB2/neuregulin signaling augments paclitaxel-induced cardiotoxicity in adult ventricular myocytes

Paclitaxel (Taxol) has been successfully combined with the monoclonal antibody trastuzumab (Herceptin) in the treatment of ErbB2 overexpressing cancers. However, this combination therapy showed an unexpected synergistic increase in cardiac dysfunction. We have studied the mechanisms of paclitaxel/anti-ErbB2 cardiotoxicity in adult rat ventricular myocytes (ARVM). Myofibrillar organization was assessed by immunofluorescence microscopy and cell viability was tested by the TUNEL-, LDH- and MTT-assay. Oxidative stress was measured by DCF-fluorescence and myocyte contractile function by video edge-detection and fura-2 fluorescence. Treatment of ARVM with paclitaxel or antibodies to ErbB2 caused a significant increase in myofilament degradation, similarly as observed with an inhibitor of MAPK-signaling, but not apoptosis, necrosis or changes in mitochondrial activity. Paclitaxel-treatment and anti-ErbB2 reduced Erk1/2 phosphorylation. Paclitaxel increased diastolic calcium, shortened relaxation time and reduced fractional shortening in combination with anti-ErbB2. A minor increase in oxidative stress by paclitaxel or anti-ErbB2 was found. We conclude, that concomitant inhibition of ErbB2 receptors and paclitaxel treatment has an additive worsening effect on adult cardiomyocytes, mainly discernible in changes of myofibrillar structure and function, but in the absence of cell death. A potential mechanism is the modulation of the MAPK/Erk1/2 signaling by both drugs.