Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Ultrastructure”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,063 records · Page 59Linked to original sources
PubMed1998

The influence of ethanol and the calcium channel antagonist, nifedipine on myocardial ultrastructure in the rat.

In this study we investigated the ultrastructural changes in rat myocardium induced by the concomitant treatment of animals with ethanol and the calcium channel antagonist, nifedipine. The animals were given perorally either placebo or 8 g/kg body weight ethanol in combination with 3 mg/kg nifedipine for 30 days. In contrast to known effects of ethanol on myocardial ultrastructure, the animals treated with ethanol and nifedipine did not have cardiomyocyte damage. The most common ultrastructural alteration was increased numbers of mitochondria. In a previous paper we reported that another calcium channel antagonist, verapamil, worsened the toxic effects of ethanol on rat myocardium. Therefore, our data could be potentially significant in the selection of a calcium antagonist for patients with increased alcohol consumption.

Animals↗
PubMed2000

Ultrastructural changes in human gingival fibroblasts in vitro after exposure to vapour phase smoke components.

Tobacco and some of its volatile and non-volatile components have been found to affect many types of cells including gingival fibroblasts. Because normal gingival fibroblast functioning is fundamental to the maintenance of the oral connective tissue as well as to wound healing, we examined the effect of two vapour phase smoke components (acrolein and acetaldehyde) on proliferation and ultrastructure of human gingival fibroblasts (HGFs) in culture. A human gingival fibroblast strain derived from healthy individuals was used in this study. The cells were incubated in the presence of different concentrations of acrolein and acetaldehyde and cell proliferation and fine morphology were evaluated. The results show that acrolein and acetaldehyde produced dose dependent inhibition of HGF viability and alteration of cytoplasmic organelles. The main ultrastructural finding for the HGF cytoplasm was the presence of vacuoles and lysosomal structures which became prominent with increasing concentration of acrolein and acetaldehyde. Our results suggest that the ultrastructural alterations we observed in HGFs may be due to the uptake and storage of acrolein and acetaldehyde by the cells.

Acetaldehyde↗
PubMed1995

Distribution and ultrastructure of somatostatin-immunoreactive cells in the pancreas of Rana esculenta.

Apart from a description of the general organization of the endocrine pancreas, the present study is focussed on the distribution and ultrastructural morphology of somatostatin-immunoreactive cells in the pancreas of the frog Rana esculenta. For light-microscopic histochemistry, the peroxidase-antiperoxidase technique was used. For the ultrastructural investigation, we employed the immunogold method. The endocrine pancreas of R. esculenta is composed of numerous islet-like structures, which contain several small somatostatin-immunoreactive cells arranged in the form of clusters. Often, however, single somatostatin cells are randomly distributed among the acinar tissue of the pancreas. These individually arranged elements possess long processes which terminate on exocrine pancreatic cells. The ultrastructural features of somatostatin-immunoreactive cells speak in favor of their endocrine and paracrine functions.

Animals↗
PubMed2003

The effect of zinc on endothelium-dependent relaxation of blood-vessels and on the ultrastructure of endothelial cells under immobilization stress.

We investigated the effects of zinc on the function and ultrastructure of endothelial cells in the case of a 48-day immobilization stress provoked in Chinchilla male rabbits (n=18) by placing them in metal hutches. Half of those rabbits (n=9) received an daily oral supplement of zinc at a dose of 0.3 mg/kg body weight (in the form of zinc acetate). The control rabbits had no intervention and received no supplement of zinc. The relaxation of smooth muscles from thoracic aorta as mediated by acetylcholine at concentrations from 10(-8) mol/L to 10(-4) mol/L was determined in isometric regime. Responses were expressed as the percentage of relaxation to prostaglandin F2alpha (2.10(-5) mol/L)-induced precontraction. The ultrastructure of endothelial cells was evaluated by electron microscopy. The level of total cholesterol and zinc in the blood serum was determined by an enzymatic method and by atomic absorption spectrometry, respectively. In rabbits receiving no zinc supplement, the relaxation of smooth muscles under the influence of acetylcholine concentrations from 10(-8) mol/L to 10(-4) mol/L was significantly (P < 0.05-0.01) lower than in rabbits receiving a supplement of zinc and lower than in control rabbits. Also, in the rabbits not receiving the zinc supplement, the level of total blood serum cholesterol was increased, but the concentration of zinc decreased. In rabbits receiving the zinc supplement, the contractility of the smooth muscles effected by acetylcholine did not change as compared with control rabbits, and we found a normal structure of endothelial cells and a normal level of total cholesterol and zinc in their blood serum. Thus, zinc played an important role in the maintenance of the normal ultrastructure and function of the endothelial cells in the rabbits receiving zinc under immobilization stress.

Acetylcholine↗
PubMed1997

Ultrastructural morphometry of parotid acinar cells following fractionated irradiation.

The aim of this study was to evaluate the long term effects on the ultrastructure of parotid glands after fractionated irradiation. The method implemented involved 5 x 6 Gy and 5 x 8 Gy, Monday to Friday 6 MV photons. By unilateral irradiation, the contralateral parotid gland served as a control. Although irradiation diminished the acinar cell density in light microscopic sections from 75 to 32% after 5 x 8 Gy of irradiation, ultrastructural morphometry could not detect any statistically significant differences in acinar cell size, nuclear size, nuclear density, granule area, mean granule size, or granule density. In general, greater differences were seen between rats receiving 30 or 40 Gy, on both the irradiated and the control size, than between the irradiated side and the control side. This was interpreted as due to differences in the nutritional state of the animals. This analysis concluded that individual acinar cells that survive irradiation seem not to be damaged in the long term when evaluated at the ultrastructural level. The study further stresses the importance of adequate sampling sizes and the use of adequate controls.

Analysis of Variance↗
PubMed2003

Effect of magnesium sulphate in experimental spinal cord injury: evaluation with ultrastructural findings and early clinical results.

Excitotoxic mechanisms have been implicated in the pathophysiology of spinal cord injury (SCI). The authors have studied the protection against secondary damage to rat spinal cord with magnesium sulphate, a well-known N-methyl-D-aspartate antagonist. Rats were randomly allocated into 5 groups. Group 1 rats were controls and normal spinal cord samples were obtained after clinical examination. 50 g-cm contusion injury was introduced to Group 2. Group 3 was vehicle, 1 cc of physiologic saline was injected post-trauma. Group 4 and 5 were treatment groups and 100 mg/kg and 600 mg/kg of Magnesium sulphate was given immediately after trauma, intraperitoneally. Animals were evaluated with inclined plane, Tarlov motor scale and Basso-Beattie-Bresnahan scale 24h after SCI. Spinal cord samples for ultrastructural evaluations were obtained following clinical examinations. Magnesium treatment improved neurological outcome. Electron microscopic results showed obvious neuroprotection in the treatment groups. Application of 600 mg/kg of magnesium revealed better ultrastructural findings and clinical results than 100 mg/kg. These findings demonstrated that magnesium sulphate possesses neuroprotection on spinal cord ultrastructure and on functional scores after acute contusion injury to the rat spinal cord.

Analgesics↗
PubMed2003

Clinical histopathology and ultrastructural analysis of myocardium following microwave energy ablation.

OBJECTIVE: Due to weaknesses of conventional modes for treating atrial fibrillation (AF), surgical energy ablation methods and tools to cure AF have been under rapid development. One of these methods, microwave energy, is beginning to be applied clinically. The purpose of this study was to examine histology and ultrastructure of lesions produced by microwave energy in the myocardium. METHODS: Fifteen consecutive patients underwent surgical microwave energy ablation (Microwave Ablation System with FLEX 4 probe, AFx Inc., Fremont, CA) concomitant to a valve procedure. Epicardial ablation was carried out on the beating normothermic heart prior to performing the valve procedure. Two tissue specimens (1cm(2)) were obtained from each patient; one from the lesion site (right appendage) and the other from an adjacent, non-ablated site, which was used as control. Tissue samples were fixed and stained as appropriate for histological and ultrastructural analysis. RESULTS: All ablated samples revealed observable microscopic alteration, including loss of nuclei, foci of coagulative necrosis or induced irregular bands of contraction. Ultrastructurally, ablated cells demonstrated architectural disarray, loss of contractile filaments, mitochondrial swelling and focal interruption of plasma membrane. CONCLUSIONS: Histologic appearance of lesions created by epicardial microwave energy ablation was consistent over tissue samples, although acute findings demonstrated differences from cryoablation. In most of the cases, lesions were transmural, as was demonstrated by loss of cellular viability throughout the depth of tissue specimens.

Atrial Fibrillation↗
PubMedDate not supplied

Impact of delayed reperfusion of myocardial hibernation on myocardial ultrastructure and function and their recoveries after reperfusion in a pig model of myocardial hibernation.

UNLABELLED: This study examined the effect of delayed reperfusion of myocardial hibernation from 24 hours to 7 days on myocardial ultrastructural and functional changes and their recoveries after reperfusion. BACKGROUND: We have previously shown in pigs that after reperfusion the functional and structural alterations in short-term myocardial hibernation which was reperfused in 24 hours can recover in 7 days. The effect of delayed reperfusion of hibernating myocardium on the extent and severity of cellular and extracellular structural changes of hibernating myocardium, and their recoveries after reperfusion is not known. METHODS AND RESULTS: A severe LAD stenosis was created in 27 pigs, reducing resting flow by 30-40% immediately after placement of the stenosis and producing acute ischemia as evidenced by regional lactate production, a decrease in regional coronary venous pH, reduced regional wall thickening (from 38.5 +/- 5.1% to 10.4 +/- 8.0%) and a 33% reduction of regional oxygen consumption. The stenosis was maintained either for 24 hours in 9 pigs (group 1) with LAD flow of 0.65 +/- 0.13 ml/min/g (38% reduction), or for 7 days in 17 pigs (group 2) with LAD flow of 0.67 +/- 0.14 ml/min/g (36% reduction). There were no differences (p = NS) in the reduction of wall thickening, rate-pressure product, lactate production, or regional oxygen consumption between group 1 and group 2. Quantitative morphometric evaluation of the ultrastructure on electromicrographs revealed a greater decrease in sarcomere volume and a higher incidence of myocytes with reduced sarcomere volume in 7-day than in 24-hour hibernating regions (53 +/- 19% versus 33 +/- 14%, p < 0.05). Patchy myocardial necrosis with replacement fibrosis was common, but 6 of the 18 pigs had no myocardial necrosis or replacement fibrosis in the 7-day hibernating group, and 4 of 9 pigs had no patchy myocyte necrosis in the 24 hour hibernating group. In 6 pigs in group 1 in which the stenosis was then released and hibernating myocardium reperfused in 24 hours, regional wall thickening recovered to 30 +/- 6% (p = NS compared to baseline) after one week of reperfusion. In 12 pigs in group 2 in which the stenosis was released and hibernating myocardium reperfused in 7 days, regional wall thickening recovered slowly, from 10.1 +/- 7.2% to 18.1 +/- 8.3% at one week (n = 5) and to 28.0 +/- 3.6% at 3-4 weeks of reperfusion (n = 7, p < 0.05 compared to baseline). Similarly, the sarcomere volume or myofilament recovered significantly (p < 0.01) and was not different compared to the normal region (p = NS) in the 24-hour hibernating region of group 1, but the recovery was much slower and was incomplete at 4 weeks (p < 0.01) compared to baseline in the 7-day hibernating region of group 2. Recovery of regional wall thickening correlated with ultrstructural recovery (p < 0.01). By multivariate stepwise regression analysis, the degree of LAD flow reduction, the extent of fibrosis, and myofilament loss were independent predictors of the extent of functional recovery. CONCLUSIONS: In a porcine model of myocardial hibernation with myocardial hypoperfusion, systolic dysfunction, and metabolic adaptations, a longer period of myocardial hibernation with delayed reperfusion was associated with more severe abnormalities of myocytes. an increasing interstitial fibrosis, and more protracted myofibrillar and functional recoveries after reperfusion. The extent of functional recovery is related to the degree of coronary flow reduction, the severity of the ultrastructural changes, and the extent of interstitial fibrosis.

Animals↗
PubMedDate not supplied

An ultrastructural study of cardiac myocytes in postmyocardial infarction ventricular aneurysm representative of chronic ischemic myocardium using semiquantitative and quantitative assessment.

To elucidate the characteristic myocytic changes in chronic ischemic myocardium, an electronmicroscopic (EM) study was carried out in surgically excised postmyocardial infarction left ventricular aneurysm (LVAN) (n = 15) using semiquantitative (in all 15 cases) and quantitative assessment methods (in 10 cases). The control group, representing normal ultrastructure, included left ventricular endomyocardial biopsy specimens of sick sinus syndrome (SSS) (n = 3), Wolff-Parkinson-White (WPW) Syndrome (n = 1), and intraoperative left ventricular endomyocardial biopsy in mitral stenosis (MS) (n = 3). Myocardial condition was assessed at the ultrastructural level according to the severity of morphologic changes, first semiquantitatively, thereafter with the use of the Image Processor-Analyzer LUZEX III for morphometric analysis. The most marked EM findings were mitochondrial regressive changes, glycogen accumulation, nuclear deformities, increased rough-surfaced endoplasmic reticulum at the perinuclear portion, lysis of myofibrils, and myofibrillar degeneration. The quantitative analysis revealed significant (p < 0.05) increase of glycogen deposition, only at the perinuclear portion in the LVAN group. The myofibril to mitochondria ratio at the intercalated disc (ID) portion of the cardiac myocytes significantly increased (p < 0.01) in the LVAN group as compared with the normal control group. Light microscopically evaluated quantitative analysis, using toluidine-blue stained semithin sections which underwent EM observation, showed that the fractional area of interstitial fibrous tissue was significantly increased in the LVAN group compared to the normal controls (p < 0.01). These results signify that in chronic ischemic myocardium, decreased consumption of glycogen in oxidative phosphorylation occurs in the surviving myocytes, and that hypertrophy of the myocytes appears. The results of this study may lead to the proper ultrastructural interpretation of biopsied human myocardium, regardless of etiology.

Adult↗
PubMed1999

Basal cell adenocarcinoma of the salivary gland: an ultrastructural and immunohistochemical study.

OBJECTIVE: The purpose of this study was to examine the ultrastructural and immunohistochemical characteristics of basal cell adenocarcinoma. STUDY DESIGN: Three cases of basal cell adenocarcinoma of the salivary glands were studied by means of light microscopy, electron microscopy, and immunohistochemistry. RESULTS: Some of the architectural tumor patterns encountered were solid, some were trabecular, and some were mixed. Ultrastructurally, solid areas were composed of nonluminal cells, some of which contained tonofilaments and well-formed desmosomes; tubulo-trabecular areas differentiated into both luminal and nonluminal cells. Both growth patterns were associated with the formation of excess basal lamina, marginally and between nonluminal cells. Myofilaments were infrequent in nonluminal cells of solid or trabecular areas. Cytokeratin (AE1/AE3) stained all 3 tumors, more peripherally in the solid pattern and usually centrally in the trabecular areas; vimentin stained all 3 tumors diffusely; smooth muscle actin (IA4) stained all 3 tumors but was mainly confined to peripheral tumor cells in both the solid and the trabecular growth patterns; epithelial membrane antigen and carcinoembryonic antigen stained 1 of the 3 tumors, predominantly in the luminal cells; p53 oncoprotein was focally positive in 2 of the 3 tumors; Ki-67 stained less than 5% of the tumor cells in all cases; and c-erb-B2 was uniformly negative in all cases. Staining patterns of cytokeratin and actin varied with the architecture of the tumor. CONCLUSIONS: Neither ultrastructural characteristics nor immunohistochemistry findings appear to distinguish basal cell adenocarcinoma from basal cell adenoma.

Actins↗
PubMed2005

[Study of mucociliary transport and nasal ciliary ultrastructure in patients with Kartagener's syndrome].

OBJECTIVE: Kartagener's syndrome (KS) is a clinical variant of primary ciliary dyskinesia involving situs inversus associated with chronic airway infections. The ciliary defect associated with this syndrome is the absence of dynein arms. The aim of this study was to evaluate mucociliary transport and ciliary ultrastructure in 14 patients with KS. PATIENTS AND METHODS: We studied nasal mucociliary transport using a radioisotopic technique and ciliary ultrastructure in 14 patients with KS. RESULTS: Thirteen patients had mucociliary stasis and 1 had severely slowed transport (1.3 mm/min). Four patients (29%) had cilia with normal dynein arms, 2 patients (14%) had short inner dynein arms, and 8 patients (57.1%) had total absence of inner and outer dynein arms. CONCLUSIONS: We conclude that the typical clinical presentation, together with altered mucociliary transport as identified by an isotopic technique, is diagnostic of KS, even when the ciliary ultrastructure is normal. KS is clinically homogenous and morphologically heterogenous.

Adolescent↗
PubMed2001

Atomic Force Microscopy and Scanning Electron Microscopy Reveal Genome-dependent Ultrastructure of Seed Surface.

Both scanning electron microscopy (SEM) and contact mode imaging via atomic force microscopy (AFM) have been utilized to elucidate the ultrastructure of mung bean seed surfaces. The results indicate: 1) that AFM is useful in the examination of seed surface ultrastructure ex-vaccuo without the need for additional complex preparative procedures; and 2) that both the cotyledon and seed coat of different strains of mung beans bear specific ultrastructural details unique to each strain. To our knowledge, these are the first AFM images of seed surfaces.

Journal Article↗
PubMed1992

Developmental changes of Echinococcus multilocularis metacestodes revealed by tegumental ultrastructure and lectin-binding sites.

Ultrastructural investigations (SEM, TEM) combined with lectin-binding analysis, have revealed concurrent modifications in tegumentary structure and surface glycoconjugates during the establishment and differentiation of Echinococcus multilocularis metacestodes in jirds. The laminated layer, which is amorphous and rich in polysaccharides when initially secreted by the young cyst, takes on a different appearance and has a different glycoconjugate composition according to whether the cyst becomes fertile or sterile. The laminated layer of fertile cysts transforms into a microfibrillar matrix, the protein content of which may increase while sugar content decreases during protoscolex differentiation. Independently of this structure, brood capsules, from which arise protoscoleces, are formed by invagination of the cyst tegument. The intense secretion of glycoconjugates from the brood capsule wall during invagination may serve to interact with host factors passing through the laminated layer. The combined use of ultrastructural study and lectin labelling has allowed the demonstration of an ultrastructural and biochemical gradient of differentiation of the protoscolex. Seven stages of differentiation have been described. The possibility that the excreted-secreted tegumentary glycoconjugates, revealed by lectin labelling during protoscolex differentiation, might be the gradual biochemical expression of one or several stimuli implicated in the phenomenon of protoscolex maturation, is discussed.

Animals↗
PubMed1997

Cloning of the entire COWP gene of Cryptosporidium parvum and ultrastructural localization of the protein during sexual parasite development.

Molecular cloning and immunoelectron microscopy have been used to clone the full-length gene encoding Cryptosporidium parvum oocyst wall protein (COWP) and to analyse at the ultrastructural level the expression and localization of COWP during development in the gut. COWP is 1622 amino acids long, has a typical leader peptide and consists of 2 amino acidic domains each containing distinct repeated elements possibly originating from a common ancestral precursor. Electron microscopy localized COWP in a large cytoplasmic inclusion and in the wall-forming bodies of early and late macrogametes, respectively. Ultrastructural analysis of double-walled sporulating and mature oocysts indicated that COWP is selectively localized in the inner layer of the oocyst wall. This study provides the first localization at the ultrastructural level of a cloned coccidian oocyst wall protein.

Amino Acid Sequence↗
PubMed1989

Albumin indirectly modulates fibrin and protofibrin ultrastructure.

Albumin modulation of fibrin and protofibrin coagulation parameters was studied. Cation-depleted, fatty acid free, human and bovine albumins decrease fibrin clot turbidity in a concentration-dependent manner. Albumin also inhibits the formation of protofibrin gels, induced by addition of 25 microM Zn(II) to protofibrils, though it does not bind to (proto)fibrin. In order to verify that competition for cations underlies the influence of albumin, fibrinogen was dialyzed against cation-depleted albumin. Elemental analysis indicates a redistribution of Zn(II) from the fibrinogen to the albumin compartment, and the resultant fibrin clots are less turbid. Apparently, cation-depleted albumin acts as a competitor for divalent cations. The ability of albumin to compete for available Zn(II) was also expressed in gels formed by pH-jump experiments, in which fibrin monomer, maintained soluble at pH 4.9, is induced to change phase by addition of NaOH to pH 7.4. While turbidimetric evidence indicates that individual fibrin fibers simply become thinner with albumin, scanning electron micrographs (SEM) reveal a more complex effect on ultrastructure. Though albumin does not bind to the gels, fibrin gels produced with albumin show major changes in fiber ultrastructure, particularly evident in gels formed in the presence of Zn(II). These structural modifications are discussed within the context of the "excluded volume" effect, in which "crowding" by albumin alters (proto)fibrin reactivity and ultrastructure.

Albumins↗
PubMed2005

Electron beam ionization induced oxidative enzymatic activities in pepper (Capsicum annuum L.), associated with ultrastructure cellular damages.

Mature green pepper fruits (Capsicum annuum L.) were subjected to ionizing radiation, in the range of 1-7 kGy, with accelerated electrons. Ultrastructural changes by electron microscopy, and the activity of several oxidative metabolism-related enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaicol peroxidase (POX), and lipoxygenase (LOX), were determined in pericarp tissue just after the ionization treatment and during postionization storage at 7 degrees C followed by 3 days at 20 degrees C. Changes in oxidative stress during the ionization treatment was assessed by the accumulation of malondyaldehide (MDA), a lipid peroxidation product. The ionization induced modifications in the cell ultrastructure, a moderate separation of the plasma membrane from the cell wall being observed for all doses. At 5 and 7 kGy, peroxisomes were not detected and the structures of the chloroplast and vacuoles were seriously damaged. Lipid peroxidation and lipoxygenase activity increased with the ionization dose, staying constant and decreasing, respectively, during the storage period. Conversely, catalase, ascorbate peroxidase, and superoxide dismutase had lower values than in nonionized fruits and, in general, their values did not change or diminished slightly from the seventh day of storage. Peroxidase exhibited an increase in activity with the ionization dose, although these was not a linear relationship, with higher values at 3kGy. Ionization of pepper, especially at doses of 5 and 7 kGy, caused a significant oxidative damage in the fruit, since it increased oxidation and decreased the antioxidant enzymatic defense systems causing ultrastructural changes at cell level.

Ascorbate Peroxidases↗
PubMed1998

Protective effects of WEB 2086 (PAF antagonist) and ketoprofen (NSAID) on PAF-induced changes in the morphological ultrastructure of blood platelets in calves.

The ultrastructure of bovine platelets was examined by transmission electron microscopy without any pretreatment (control), and after WEB 2086 (a triazolodiazepine) or ketoprofen (NSAID) pretreatment, followed by PAF infusion. The blood platelet count was also investigated. The group of calves that received WEB 2086 pretreatment before platelet-activating factor (PAF) infusion did not show a decreased number of platelets. However, in the other group, with ketoprofen pretreatment before PAF infusion, there was a rapid decrease from 1 to 3 min, while from 5 min the number of platelets recovered to the normal value. Electron microscopy revealed that pretreatment with WEB 2086 followed by PAF infusion did not alter the morphological ultrastructure of bovine platelets, except that the microtubules were briefly modified from 1 until 3 min after PAF challenge. After ketoprofen pretreatment, bovine platelets kept their regular shape, the number of dense bodies was not significantly altered, the number of mitochondria was maintained from 5 min after PAF infusion, giant platelets were not observed and the Golgi apparatus was rarely visible. Thus pretreatment with WEB 2086 and ketoprofen before PAF infusion had a protective activity on the ultrastructure of bovine platelets and, in cattle, pretreatment with WEB 2086 and ketoprofen before PAF challenge prevented the thrombocytopenia induced by PAF.

Animals↗
PubMed2000

Altered transarcolemmal Ca transport modifies the myofibrillar ultrastructure and protein metabolism in cultured adult ventricular cardiomyocytes.

The present study was designed to investigate how prolonged (24-72 h) exposure to modifiers of Ca transarcolemmal transport affects the myofibrillar structure, protein turnover and content of myofibrillar proteins in adult guinea pig cardiomyocytes maintained beating synchronously in long-term cultures. First we established the functional responses (the contractile activity and [Ca]i transients) of the cultured myocytes to acute exposures to several drugs used in this study. The ultrastructural characteristics of these cultures under the various treatments were determined using immunohistochemistry and confocal scanning laser microscopy, and their biochemical properties were evaluated using analysis of total cellular protein content, myofibrillar protein content and SDS-PAGE electrophoretic examination. We compared the effects of 24, 36 and 72 h-long exposures to the various specific Ca-flux modifiers. Increased Ca influx via CaL-channel agonist (Bay K 8644) or via the reversed-mode of the Na/Ca exchanger (veratrine) did not alter the myofibrillar structure or the specific protein profiles or proteosynthesis. However, when cytosolic Ca was increased by three different types of inhibitors of Ca extrusion from the cells via Na/Ca exchange, (Na-free solution, 5 mM NiCl2 and 10(-6) M ouabain), very significant changes in all investigated parameters occurred almost immediately. Twenty-four h-long exposure to Na-free did not affect significantly the total cellular protein (TCP), but the protein synthesis was decreased by 87% and the total myofibrillar protein (TMP) content was decreased by 38%. The myofibrils were heavily fragmented. Similarly, 24 h-long exposure to 5 mM NiCl2 did not affect the TCP, but it reduced protein synthesis by about 90% and decreased the total myofibrillar protein content by 30%. These effects were even more pronounced at 72 h of exposure and they were accompanied with a complete disassembly of myofilaments. Exposure to 10(-6) M ouabain over 72 h resulted in > 80% inhibition of protein synthesis, a 45% decrease in TCP content and a 53% in TMP content. In contrast, 10(-7) M ouabain did not produce any such changes. The changes produced by the Na/Ca-exchange inhibitors were accompanied by only minor changes in DNA content, indicating that the myocytes remained viable. Moreover, these effects were not due to the associated contractile arrest, since exposure to CaL-channel antagonists (5-20 microM nifedipine or 10 microM verapamil) produced only very minor changes in the myofibrillar structure and in protein profiles. Our data demonstrate that short-term (up to 72 h) increased Ca influx or contractile arrest of well-interconnected, spontaneously beating adult cardiomyocytes does not affect their ultrastructural characteristics or their myofibrillar protein turnover greatly, while any situations leading to Ca accumulation (via inhibition of Na/Ca exchange) affect cardiomyocyte function and ultrastructure almost immediately. These data are in sharp contrast to those previously reported from immature, neonatal myocytes.

Animals↗