Carnitine derivatives in hereditary cardiomyopathic animals

BIO 14.6 Syrian hamsters and diabetic KK mice have been reported to develop hereditary cardiomyopathy spontaneously. In order to investigate the pathophysiological role of carnitine metabolism in hereditary cardiomyopathy, tissue levels of carnitine derivatives and the histology of the heart, liver and skeletal muscles from BIO 14.6 hamsters and KK mice were studied. Free carnitine levels in the heart of the BIO 14.6 hamsters (287.0 +/- 27.0 n mole/g wet tissue) were significantly lower than in the control group (348.8 +/- 83.8, p less than 0.05). Short chain acylcarnitine (197.0 +/- 56.0 n mole/g wet tissue) and total carnitine (667.6 +/- 136.4 n mole/g wet tissue) in the hearts of the BIO 14.6 hamsters were significantly lower than in the control group (short: 425.2 +/- 54.8, total: 1023.6 +/- 81.4, p less than 0.001). There was no significant difference in the levels of various carnitine derivatives of the liver and skeletal muscles from the BIO 14.6 hamsters and control hamsters. On the other hand, carnitine derivatives in KK mice did not change significantly compared with those in the heart, liver and skeletal muscles of the control mice. Histological findings showed that heart muscle degeneration and necrosis were found in both cardiomyopathic animals. Coagulative necrosis was found in both animals, whereas myocytolytic necrosis was found only in the BIO 14.6 hamsters. In KK mice, the right ventricle, especially tissue under the epicardium, was severely affected compared with the left ventricle. In the BIO 14.6 hamsters, however, lesions were scattered over both ventricles with a predilection for the left ventricle.     

The effect of ouabain on hearts of cardiomyopathic hamsters: potentiation by isoproterenol

The isometric twitch properties of papillary muscles from hearts of 30- to 53-day-old cardiomyopathic hamsters (BIO 14.6) were studied before and after exposure to the cardiac glycoside, ouabain. The diseased tissue was weakly responsive to ouabain (3 to 100 microM), as compared to a more appreciable positive inotropic response in papillary muscle of similarly aged normal hamsters (BIO F1B). These data suggest that the activity of Na+,K+-ATPase is attenuated in the diseased sarcolemma. Pretreatment with the beta-adrenoceptor agonist, isoproterenol (0.1 microM), slightly increased the sensitivity of normal muscle to ouabain, however the response of myopathic muscle was greatly enhanced. These findings may be of significance to the genesis of cellular calcium overload hypothesized to be involved in the necrosis and degeneration of heart cells in this animal model of genetic cardiomyopathy.     

Isoprenaline-evinced disturbances in action potentials from hearts of young cardiomyopathic hamsters

The electrophysiology of ventricular cells from prenecrotic stage 10 to 14 day old hamsters with hereditary cardiomyopathy (BIO 14.6 strain) was studied with the intent of learning more about the previously documented relationship of stimulation with beta-adrenergic agonist and the sarcolemmal defect contributing to the excessive uptake of calcium by diseased cells. Before catecholamine treatment only slight differences were observed in the configuration of action potentials of myopathics and control random bred hamsters (BIO RB strain). However, isoprenaline in varying concentrations, increased the action potential duration (APD) at 50% and 95% repolarisation levels to a significantly greater extent in myopathics than in controls. Repetition of the dose-response to isoprenaline in the presence of the beta-adrenergic antagonist propranolol (0.1 mumol X litre-1) permitted the calculation of the dissociation equilibrium constant for the antagonist. The similarity of dissociation constants between strains, 0.008 mumol X litre-1 for controls and 0.011 mumol X litre-1 for myopathics, suggests that a difference in interstrain receptor affinities is an unlikely cause of the isoprenaline effect. Rather, the data are more consistent with the hyper-sensitivity resulting from larger numbers of beta-adrenoceptors on sarcolemma of myopathic cells. Also, the increase in APD of myopathics indicates that isoproterenol elicits an imbalance of slow inward calcium and late outward potassium currents. The possible significance of the isoprenaline hyper-sensitivity, which is the earliest pathophysiology seen in this disease, to the etiology of cellular calcium overload and degeneration of diseased myocardium remains to be determined.     

Functional state of myofibrils, mitochondria and bound creatine kinase in skinned ventricular fibers of cardiomyopathic hamsters

Functional states of cardiac contractile apparatus and mitochondria were studied in hereditary cardiomyopathic hamsters (CHF 146) and control golden hamsters using cardiac fibers skinned by two different techniques. The Triton X-100 skinned fibers obtained from diseased animals of 175 to 200 days old, or from control animals, demonstrated the same resting and maximal Ca-activated tensions, the same stiffness, the same rate of tension recovery after quick stretch; the fibers from cardiomyopathic animals differed only by a slightly increased calcium sensitivity. Functional activity of myofibrillar creatine kinase in cardiomyopathy was decreased as indicated by a smaller shift in the pMgATP/rigor tension curve to lower [MgATP] in the presence of phosphocreatine and by a slower rate of the tension recovery after quick stretch in the presence of phosphocreatine and ADP (without ATP). The saponin-skinned fibers allow evaluation of the respiration properties of the total tissue mitochondria. Data obtained in the preparations isolated from diseased animals of two ages (75 to 100 and 175 to 200 days) showed that the ratio of maximal ADP-stimulated respiration rate to the respiration rate in the absence of ADP (an analog of respiration control index) was unchanged in myopathy as compared with age-matched controls. However stimulation of respiration after an addition of creatine at submaximal ADP concentration was observed to be respectively 1.45 times and 3.5 times less in the preparations from younger and older myopathic animals as compared with their respective controls, thus indicating the impairment of functional coupling between mitochondrial creatine kinase reaction and oxidative phosphorylation. These results suggest that hereditary cardiomyopathy is associated with alterations in myocardial creatine kinase system, while myofilaments and mitochondria preserve their basic functional properties.     

Diminished beta-adrenergic receptor responsiveness and cardiac dilation in hearts of myopathic Syrian hamsters (BIO 53.58) are associated with a functional abnormality of the G stimulatory protein

Previous studies have demonstrated a diminution in the bioactivity of the guanine nucleotide-binding regulatory protein that stimulates adenylyl cyclase (Gs) in hearts of the hypertrophic BIO 14.6 Syrian hamster. In this study, we measured functional activity and immunodetectable levels of Gs in a mutant strain of hamsters (BIO 53.58) that develop a dilated cardiomyopathy. Pathological studies demonstrated that 100-day-old BIO 53.58 hamsters had substantial ventricular dilation when compared with age-matched F1B controls. Additionally, these 100-day-old hamsters demonstrated diminished contractile response to beta-adrenergic receptor stimulation. The pathological and hemodynamic changes were associated with defective coupling of Gs to adenylyl cyclase as adenylyl cyclase activation was distinctly decreased in the presence of isoproterenol, fluoride ion, guanine nucleotides, and forskolin. Additionally, the ability of the alpha-subunit of Gs to reconstitute isoproterenol-stimulated adenylyl cyclase activity in S49 cyc- membranes was reduced approximately 65%. By contrast, cyc- complementation assays did not reveal a difference between the functional activity of Gs in hearts from 30-day-old BIO 53.58 hamsters and F1B controls. Furthermore, beta-adrenergic receptor stimulation of adenylyl cyclase in the membranes of the young BIO 53.58 hamsters was not significantly different from controls. The substantial alterations in Gs bioactivity in hearts of the 100-day-old BIO 53.58 hamsters was not associated with alterations in the immunodetectable levels of either alpha Gs or alpha Gi on Western Blots. These results suggest that G protein changes are associated with ventricular dilation in BIO 53.58 hamsters and that G protein levels are not always reflective of G protein bioactivity.     

Does an impaired adenosine mediated feedback control play a role in the development of hereditary dystrophic cardiomyopathy?

A study was carried out to investigate whether or not an impairment of the adenosine mediated negative inotropic effect in the presence of beta adrenoceptor stimulation plays a role in the pathogenesis of the hereditary cardiomyopathy of the Syrian hamster. In electrically driven papillary muscles isolated from the hearts of cardiomyopathic (strain BIO 8262) and age matched healthy control Syrian hamsters the effects of isoprenaline, adenosine, and adenosine in the presence of isoprenaline were studied within the first 30 days of life (the prenecrotic stage of the disorder). In both cardiomyopathic and control hamsters adenosine antagonised the positive inotropic effect of isoprenaline, whereas adenosine alone had no or, only a weak, inhibitory effect on the force of contraction. The effects in both groups were similar. The effect of isoprenaline on the force of contraction also did not differ in the two groups. The data show that in both cardiomyopathic and control hamsters adenosine reduces the force of contraction during beta adrenergic stimulation. The potency or efficacy of adenosine did not differ in the two groups. An impaired adenosine mediated feedback control of the heart does not therefore seem to play a role in the pathogenesis of the hereditary dystrophic cardiomyopathy of the Syrian hamster.     

Calcium sensitivity and myosin light chain pattern of atrial and ventricular skinned cardiac fibers from patients with various kinds of cardiac disease

In the present study, the Ca2(+)-sensitivity and myosin light chain patterns of skinned fibers of right atrium and left papillary muscles of 27 patients suffering from mitral valve disease (MVD, moderate heart failure), ischemic cardiomyopathy (ICM, severe heart failure), dilated cardiomyopathy (DCM, severe heart failure), and coronary heart disease (CHD, no heart failure, no atrial hypertrophy) were investigated. Myosin light chains of both chemically skinned and intact samples were studied by two-dimensional gel electrophoresis (2D-PAGE). Ca2(+)-sensitivity of ventricular fibers was about 0.14 pCa-units higher than that of atrial fibers in all groups except dilated cardiomyopathy where this difference was markedly diminished (only 0.06 pCa-units). Generally, Ca2(+)-sensitivity of skinned ventricular fibers was the same among the different heart diseases. Skinned atrial fibers from patients with dilated cardiomyopathy, however, were significantly (about 0.08 pCa-units) more sensitive for Ca2+ than those of the other groups (coronary heart disease, mitral valve disease or ischemic cardiomyopathy) which showed similar Ca2(+)-tension relationships. Ventricle-specific P-light chain forms could be observed in atrial samples from patients of all groups, whereas no atrium-specific light chain forms were detectable in any ventricular sample. It is concluded that there is no difference in Ca2(+)-sensitivity of the ventricular contractile elements of the human heart in different heart diseases. In atrial myocardium, there is an increased Ca2(+)-sensitivity of skinned fibers from hearts with dilated cardiomyopathy which is probably related to an elevation of right atrial pressure.     

Increased left ventricular diastolic stiffness in the early phase of hereditary cardiomyopathy

Isolated hearts from normal and cardiomyopathic hamsters (160 to 180 days of age) were perfused through the aorta and assessed by echocardiographic and 31P-NMR (nuclear magnetic resonance) techniques. A decreased left ventricular systolic pressure in cardiomyopathic hamsters was associated with diminished cardiac size and left ventricular wall thickness. However, the ratio of inner/outer cross-sectional area and estimated left ventricular volume at any given left ventricular weight was significantly higher, indicating relative left ventricular chamber enlargement in cardiomyopathic hamsters. Left ventricular volumes were increased with an intraventricular balloon. Gradual inflation of the balloon resulted in increments of left ventricular systolic and developed stress that rose to the same values in both groups. At this point, the normalized stress-strain relationship was approximately two times steeper for cardiomyopathic hamsters, while at lower strain values the diastolic stress in cardiomyopathic hamsters was less than in controls, possibly due to cardiac dilatation. Almost the same degree of dilatation was induced in control hearts by the acute addition of 1% alcohol, but it was not followed by increased diastolic stiffness. Examination of hearts by 31P-NMR techniques revealed a decreased phosphocreatine/inorganic phosphate (PCr/Pi) ratio in the cardiomyopathic hamsters that progressed further with balloon inflation and was associated with a relative fall in PCr and adenosine triphosphate (ATP) content. Results suggest increased diastolic stiffness in cardiomyopathic hamsters, which was not seen in acute cardiac depression with alcohol. Diastolic volume overload with increased wall stress is probably the major factor contributing to increased diastolic stiffness early in the cardiomyopathy.     

Possible involvement of free radicals and antioxidants in the early stages of the development of cardiomyopathy in BIO 14.6 Syrian Hamster.

The BIO 14.6 cardiomyopathic Syrian hamster is a well-known animal model of congestive cardiomyopathy. To evaluate the role of free radicals and antioxidant protection in the pathogenesis of cardiomyopathy in this animal, we studied the concentration of heart mitochondrial free radicals, the activities of glutathione peroxidase (GSHPx) and superoxide dismutase (SOD), and the effect of alpha-tocopherol on the early stage of myocardial damage (up to 90 days). The GSHPx activity in BIO 14.6 hamsters was found to be twice that in the normal control hamsters at 30 days of age, while SOD activity was unchanged at 30 and 90 days of age. The concentrations of mitochondrial free radicals in BIO 14.6 hamsters at 40 and 90 days of age were significantly higher than those in the normal control hamsters. A protective effect of alpha-tocopherol therapy was shown in BIO 14.6 hamsters treated during the early stage of cardiomyopathy (up to 90 days). These results show the role of free radicals and antioxidant protection in the pathogenesis of hamster cardiomyopathy. We suspect that an increase in the GSHPx activity in BIO 14.6 hamsters may be due to a compensatory mechanism to counteract oxidative stress, but antioxidant reserve was not sufficient to protect the heart from the toxic effects of increased free radicals in the early stage of cardiomyopathy.     

Absence of Bowditch phenomenon in the ventricular muscle of hamsters with hereditary cardiomyopathy

The interval-tension relationship of right ventricular muscles of normal hamsters was compared to that of hamsters with hereditary cardiomyopathy, with or without congestive heart failure. The contractility of the heart of animals with cardiomyopathy without circulatory insufficiency did not differ from that of the normal animals at any frequency. The cardiac muscle of animals with severe congestive heart failure showed normal contractility at a low frequency of stimulation (range of the Woodworth phenomenon) but did not show the increased contractility upon high frequency stimulation (Bowditch phenomenon) seen in all other animals. Ryanodine, which eliminates only the contractility caused by the Woodworth phenomenon, abolished the contractility of the hearts of animals with congestive failure over the whole frequency range. Cardiac glycosides, which are known to potentiate the Bowditch phenomenon, were ineffective on the cardiac muscle of animals without Bowditch phenomenon. The probable cause of cardiac failure in hamsters with hereditary cardiomyopathy is discussed.     

Diminished effects of L-thyroxine on some functions and enzymes of the heart in myopathic hamsters.

The effects of chronic administration of l-thyroxine on some functions of the heart in normal and myopathic Syrian hamsters (strain BIO 14.6) were investigated. In contrast to normal controls, in myopathic hamsters administration of low doses of l-thyroxine for 4 weeks failed to induce tachycardia, increase the total activity of NaK-ATPase (EC 3.6.1.3) within the cells or improve contractility of the heart muscle. In addition, l-thyroxine treatment decreased the activity of adenyl cyclase in the myopathic and that of soluble protein kinases in both normal and myopathic hearts. Some mechanism(s) that mediate the action of l-thyroxine in the myocardium therefore may be altered or deficient in this form of hereditary cardiomyopathy of the hamster.     

Opposing effect of p38 MAP kinase and JNK inhibitors on the development of heart failure in the cardiomyopathic hamster

OBJECTIVE: p38 MAP kinase (p38 MAPK) and c-Jun NH2-terminal kinase (JNK) have been implicated in the pathophysiology of heart failure. We investigated the effects of chronic treatment with p38 MAPK and JNK inhibitors on the development of heart failure in dilated cardiomyopathy (DCM) hamster heart. METHODS AND RESULTS: BIO14.6 hamster hearts showed markedly increased p38 MAPK and JNK activities at 6 weeks of age when there was no significant increase in the area of fibrosis, heart weight/body weight, left ventricular (LV) chamber dilation and LV dysfunction. p38 MAPK and JNK activities were attenuated at 26 weeks of age and abolished at 40 weeks of age in BIO14.6 hamster hearts. BIO14.6 hamsters and the control BIOF1B hamsters were chronically treated (i.p.) with the p38 MAPK inhibitors, SB203580 (1 mg/kg/day) and FR167653 (3 mg/kg/day), or the JNK inhibitor, SP600125 (1 mg/kg/day) or vehicle for 20 weeks starting from 6 weeks of age. Treatment of BIO14.6 hamster hearts with SB203580 and FR167653 reduced the number of TUNEL-positive myocytes, the area of fibrosis and heart weight/body weight associated with a significant decrease of LV dimension and an increase in LV ejection fraction and LV contractility compared to the vehicle-treated counterpart. In contrast, treatment with SP600125 increased the number of TUNEL-positive myocytes and the area of interstitial fibrosis associated with aggravation of LV chamber dilation and LV dysfunction. CONCLUSIONS: These results suggest that chronic treatment with p38 MAPK and JNK inhibitors produces opposing effects on the development of heart failure in the DCM hamster heart.     

3H]-nitrendipine binding sites in normal and cardiomyopathic hamsters: absence of a selective increase in putative calcium channels in cardiomyopathic hearts

The number of putative calcium channels in cardiac muscle from young adult hamsters (60 days old) was compared in normal (F1B) hamsters and two different mutant strains (CHF 146 and Bio 14.6) which express cardiomyopathy and muscular dystrophy. Equilibrium binding assays of high affinity sites for [3H]-nitrendipine in ventricular homogenate preparations showed that the maximum number of [3H]-nitrendipine binding sites (Bmax), which corresponds to the number of putative calcium channels, was not significantly different in normal and cardiomyopathic hearts: 79(SEM 9), 64(14) and 69(10) fmol.mg-1 protein in 4-6 hearts from F1B, Bio 14.6 and CHF 146 hamster strains, respectively. Similar results were obtained with binding data after partial purification of the preparation. These data are in agreement with earlier studies comparing two normal strains (CHF 148 and random bred Syrian hamsters) with cardiomyopathic (CHF 146) hamsters, and conflict with other studies comparing normal and cardiomyopathic hamsters. Comparisons with the conflicting data suggest (a) that change in the number of high affinity [3H]-nitrendipine binding sites is not responsible for calcium overload and cell necrosis in cardiomyopathy, and (b) that increased numbers of low affinity [3H]-nitrendipine binding sites may emerge in cardiomyopathic hearts.     

Value of the isoprenaline test in arrhythmogenic heart diseases

The sympathetic nervous system seems to play a major role in the genesis of ventricular arrhythmias. The authors studied this adrenergic factor prospectively by exercise stress testing and intravenous isoprenaline in 107 patients referred for evaluation of arrhythmias or symptoms thought to be due to arrhythmias: 30 patients had morphologically normal hearts (15 ventricular extrasystoles, 15 bursts of ventricular tachycardia); 55 patients had dilated cardiomyopathy and 22 had probable or proven arrhythmogenic right ventricular dysplasia. Exercise testing was carried out with 30 watt increments every 3 minutes. Ventricular tachycardia was induced in 6 patients with apparently normal hearts (17%), 13 patients with dilated cardiomyopathy (31%) and 7 patients with arrhythmogenic right ventricular dysplasia (40%). Isoprenaline was infused for 3 minutes at a dose of 8-12 g/min: ventricular tachycardia was induced in 7 patients with apparently normal hearts (24%) and 23 patients with dilated cardiomyopathy. In some patients presenting with syncope, an arrhythmogenic response to isoprenaline was the only abnormality detected by the study protocol. An arrhythmia was induced by isoprenaline in 17 of the 18 patients with confirmed right ventricular dysplasia (94%), 12 of whom had sustained mono or polymorphic ventricular tachycardia. Two of these patients did not have significant right ventricular wall motion abnormalities. Four asymptomatic subjects related to patients with right ventricular dysplasia underwent the isoprenaline test; bursts of ventricular tachycardia were recorded in 3 of them. Polymorphic ventricular tachycardia was specifically associated with cardiac disease. The maximum heart rate attained by exercise testing (148 +/- 19/min) was higher than that attained with isoprenaline (148 +/- 22/min).(ABSTRACT TRUNCATED AT 250 WORDS)     

Contractile properties of papillary muscle from young cardiomyopathic hamsters: effects of isoprenaline

Papillary muscles isolated from the left ventricle of 17-20 day old Syrian hamsters with genetic cardiomyopathy (BIO 14.6 strain) were examined for their mechanophysiological response to the beta adrenoceptor agonist isoprenaline. The indices of isometric twitch for both myopathic and aged matched normal (BIO RB strain) hamster papillary muscles were statistically the same before exposure to catecholamine. Isoprenaline, however, elicited an exaggerated degree of positive inotropism in myopathic papillary muscles. In addition, the characteristic relaxing effect of beta adrenoceptor agonists (abbreviation of twitch duration) was virtually absent in myopathic muscles. These results suggest that under conditions resembling sympathetic stimulation sarcolemma and perhaps sarcoplasmic reticulum become inadequate regulators of calcium, predisposing young myopathic heart cells to accrue damaging concentrations of sarcoplasmic calcium.     

Role of alpha 1-adrenergic receptors and the effect of bunazosin on the histopathology of cardiomyopathic Syrian hamsters of strain BIO 14.6

In order to clarify the pathological involvement of the sympathetic nervous system in the development of cardiomyopathy, a receptor-binding study was carried out on cardiomyopathic Syrian hamsters of strain BIO 14.6 (BIO) at 21 days (prenecrotic stage); 35-42 days (onset of cardiomyopathy); and 70-84 days of life (early cardiac hypertrophy). The newly developed alpha 1-blocker (bunazosin hydrochloride) was initially administered at doses of 100 micrograms/kg or 10 mg/kg to BIO hamsters at 21 days of life and continued for 70 days. At the onset of cardiomyopathy and early cardiac hypertrophy, there was an increase in the number of alpha 1-receptors in the BIO hamsters compared to controls, but there were no significant changes at the prenecrotic stage. On histopathological examination, 10 mg/kg bunazosin had a significant beneficial effect on cardiomyopathy [area of necrosis 1.38% in untreated vs 0.33% in treated animals; area of calcification 2.70% (untreated) vs 0.60% (treated); area of all myocardial injuries 6.97% (untreated) vs 3.19% (treated)]. However, 100 micrograms/kg bunazosin had no effect. It was concluded that the increase in the number of alpha 1-receptors may not be involved in the pathogenesis of cardiomyopathy but that alpha 1-receptors could be implicated in the later progression of the condition.     

Contractile properties and Ca2+ release activity of the sarcoplasmic reticulum in dilated cardiomyopathy.

BACKGROUND. We performed a comparative study on Ca2+ release activity of the sarcoplasmic reticulum and calcium sensitivity of contractile apparatus of chemically skinned myocardial fibers obtained from four nonfailing human hearts and 13 excised hearts from patients with idiopathic dilated cardiomyopathy. METHODS AND RESULTS. Ca2+ sensitivity of contractile apparatus was studied by following the isometric tension developed by chemically skinned myocardial fibers challenged with solutions of decreasing pCa. Ca2+ release from sarcoplasmic reticulum was monitored indirectly by measurement of the isometric tension developed by skinned fibers challenged with caffeine. We observed no significant difference of Ca2+ sensitivity and cooperativity between normal myocardium (pCa50 = 6.00 +/- 0.05; Hill coefficient, nHill = 2.07 +/- 0.10) and dilated cardiomyopathy (pCa50 = 6.03 +/- 0.07; nHill = 2.72 +/- 0.30) when the fibers were stretched to 130% of the resting length. We also found that both in normal myocardium and dilated cardiomyopathy, stretching to 150% of the resting length increased the Ca2+ sensitivity of the contractile system; pCa50 = 6.21 +/- 0.01 and 6.13 +/- 0.04 in normal and dilated cardiomyopathy, respectively, whereas in dilated cardiomyopathy there was a decrease of Hill coefficient with stretching that was not observed in the control group. The caffeine threshold in idiopathic dilated cardiomyopathy was markedly increased compared with the control group, 1.94 +/- 0.27 mmol/l and 0.29 +/- 0.04 mmol/l caffeine, respectively, whereas there were no significant differences in the extent and rate of caffeine-induced Ca2+ release. CONCLUSIONS. These results indicate that in idiopathic dilated cardiomyopathy there is no alteration of contractile and regulatory proteins; on the contrary, the gating mechanism of the Ca2+ release channel of sarcoplasmic reticulum is abnormal, suggesting a possible involvement of the excitation-contraction coupling in the pathogenesis of this disease. It should also be taken into account, however, that the increased caffeine threshold in dilated cardiomyopathy would be a result of the enhanced resistance to the skinning procedure secondary to the modification of lipid species and/or content in sarcoplasmic reticulum membrane.     

The studies of cell damaging and cell growth factors which induce cardiomyopathy.

We demonstrated that phosphatidylinositide-specific phospholipase C (PLC) activity was greater in cardiomyopathic hamster hearts (BIO 14.6 and BIO 53.58) then in hamster controls (F1b). Inositol trisphosphate (IP3) production was markedly greater in both of the cardiomyopathic hamsters, BIO 14.6 and BIO 53.58. We have also determined the sarcoplasmic reticulum (SR) function of heart. Calcium uptake into SR markedly increased in BIO 14.6. On the other hand, it significantly decreased in BIO 53.58 compared with F1b. It is well known that IP3 stimulates calcium release from SR. In BIO 14.6, calcium release from SR stimulated by IP3 increased, but its effect decreased in BIO 53.58 compared with F1b. These results suggest that PI response may produce high intracellular calcium levels in both BIO 14.6 and BIO 53.58 myocytes. In addition, in the BIO 53.58 hamster the sarcoplasmic reticulum deteriorate in function. It was concluded from these results that a prolonged high intracellular calcium level may lead to the death of BIO 53.58 myocytes. The expression of angiotensinogen mRNA was observed in the hamster heart. There was no differences in its expression level between F1b, BIO 14.6 and BIO 53.58. There was no effect of ages on its expression in these hamster hearts. We have also determined the distribution of angiotensinogen in these hamsters. At 4 weeks of age, the immunohistochemical study revealed that angiotensinogen was widely distributed in subendocardium in these hamsters. There was no difference in its distribution between F1b, BIO 14.6 and BIO 53.58. But at 20 weeks old of age its immunoreactivity decreased in BIO 53.58.(ABSTRACT TRUNCATED AT 250 WORDS)     

Decreased 1,2-diacylglycerol levels in myopathic hamster hearts during the development of heart failure

1,2-Diacylglycerol is believed to play an important role in cellular functions through protein kinase C activation, although its role in cardiac functions remains largely unexplored. We determined the level of 1,2-diacylglycerol and its fatty acid composition in heart tissues from Syrian hamsters with hereditary cardiomyopathy (BIO 14.6 strain) during the development of congestive heart failure from 90 days to 240 days of age. The myopathic hamsters had lower contents of triglyceride and of the major phospholipids, phosphatidylcholine, phosphatidylethanolamine and cardiolipin, in the myocardium when compared to normal hamsters, whereas there was no difference in the cholesterol content. No difference in the myocardial 1,2-diacylglycerol content was observed at 90 days of age. On the other hand, 1,2-diacylglycerol contents in myopathic hearts at 160 and 240 days of age were significantly lower by 21% and 52%, respectively, then in age-matched normal hamsters. The oldest hamsters (240-day-old) showed reduced 1,2-diacylglycerol levels in both groups despite an age-related increase in most lipids. The 1,2-diacylglycerol fatty acid composition profile was found to be different from that of other lipids, and there were several differences in the fatty acid composition of 1,2-diacylglycerol between the two groups at 240 days of age. These results indicate that decreased levels of 1,2-diacylglycerol occur concomitantly with congestive heart failure in the myopathic hamsters.