• Proceedings of the Korean Biophysical Society Conference
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• 1999.06a
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• pp.53-53
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• 1999

We have investigated whether alterations in the expression levels of sarcoplasmic reticulum (SR) $Ca^{2＋}$ regulatory proteins in heart and mesenteric arteries from different models of hypertension would occur. Nephrectomied diabetic-hypertensive rats (DM-HT) and spontaneously hypertensive rats (SHR) were used as models of hypertension.(omitted)

• Proceedings of the Korean Biophysical Society Conference
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• 1997.07a
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• pp.18-18
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• 1997

Chronic treatment with cyclosporin A (CsA) were shown to induce reversible alterations of contractile properties in rat heart. To define the molecular mechanisms underlying the physiological alterations, the $Ca^{2＋}$ release channel (CRC) and $Ca^{2＋}$-ATPase in rat sarcoplasmic reticulum (SR) were examined.(omitted)

• Proceedings of the Korean Biophysical Society Conference
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• 1997.07a
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• pp.30-30
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• 1997

Dysfunctions of skeletal muscles have been frequently reported in chronic diabetic mellitus (DM). In order to investigate the molecular mechanisms of abnormal function, the junctional sarcoplasmic reticulum (HSR) vesicles of skeletal muscles were prepared from the control and the streptozotocin-induced diabetic rats.(omitted)

• Molecules and Cells
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• v.20 no.1
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• pp.51-56
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• 2005

Excitation-contraction coupling (ECC) proteins in the human heart were characterized using human atrial tissues from different age groups. The samples were classified into one infant group (Group A: 0.2-7 years old) and three adult groups (Group B: 21-30; Group C: 41-49; Group D: 60-66). Whole homogenates (WH) of atrial tissues were assayed for ligand binding, $^{45}Ca^{2+}$ uptake and content of ECC proteins by Western blotting. Equilibrium [$^3H$]ryanodine binding to characterize the ryanodine receptor (RyR) of the sarcoplasmic reticulum (SR) showed that the maximal [$^3H$]ryanodine binding ($B_{max}$) to RyR was similar in all the age groups, but the dissociation constant ($k_d$) of ryanodine was higher in the infant group than the adult groups. Oxalate-supported $^{45}Ca^{2+}$ uptake into the SR, a function of the SR SERCA2a activity, was lower in the infant group than in the adult groups. Similarly, [$^3H$]PN200-110 binding, an index of dihydropyridine receptor (DHPR) density, was lower in the infant group. Expression of calsequestrin and triadin assessed by Western blotting was similar in the infant and adult groups, but junctin expression was considerably higher in the adult groups. These differences in key ECC proteins could underlie the different $Ca^{2+}$ handling properties and contractility of infant hearts.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.4
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• pp.195-201
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• 2005

High extracellular glucose concentration was reported to suppress intracellular $Ca^{2+}$ clearing through altered sarcoplasmic reticulum (SR) function. In the present study, we attempted to elucidate the effects of pyruvate and fatty acid on SR function and reveal the mechanistic link with glucose-induced SR dysfunction. For this purpose, SR $Ca^{2+}$-uptake rate was measured in digitonin-permeabilized H9c2 cardiomyocytes cultured in various conditions. Exposure of these cells to 5 mM pyruvate for 2 days induced a significant suppression of SR $Ca^{2+}$-uptake, which was comparable to the effects of high glucose. These effects were accompanied with decreased glucose utilization. However, pyruvate could not further suppress SR $Ca^{2+}$-uptake in cells cultured in high glucose condition. Enhanced entry of pyruvate into mitochondria by dichloroacetate, an activator of pyruvate dehydrogenase complex, also induced suppression of SR $Ca^{2+}$-uptake, indicating that mitochondrial uptake of pyruvate is required in the SR dysfunction induced by pyruvate or glucose. On the other hand, augmentation of fatty acid supply by adding 0.2 to 0.8 mM oleic acid resulted in a dose-dependent suppression of SR $Ca^{2+}$-uptake. However, these effects were attenuated in high glucose-cultured cells, with no significant changes by oleic acid concentrations lower than 0.4 mM. These results demonstrate that (1) increased pyruvate oxidation is the key mechanism in the SR dysfunction observed in high glucose-cultured cardiomyocytes; (2) exogenous fatty acid also suppresses SR $Ca^{2+}$-uptake, presumably through a mechanism shared by glucose.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.6
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• pp.707-716
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• 1997

Recent reports revealed that the $Na^+-Ca^{2+}$ exchangers and feet structures of sarcoplasmic reticulum(SR) are located in close vicinity in the specific compartment. Therefore, we investigated the possibility that the $Na^+-Ca^{2+}$ exchanger may decrease the tension development by transporting the $Ca^{2+}$ out of the cell right after it released from SR, on the basis of this anatomical proximity. We exammined the negative force-frequency relationship of the developed tension in the electrically field stimulated left atria of postnatal developing rat(1, 3 day, 1 week and 4 week old after birth). Cyclopiazonic $acid(3{\times}10^{-5}\;M)$ treatment decreased the developed tension further according to postnatal age. $Monensin(3{\times}10^{-6}\;M)$ treatment did not increase the maximal tension in 4 week-old rat, preserving negative staircase, while the negative staircase in the younger rat were flattened. $Ca^{2+}$ depletion in the buffer elicited more suppression of the maximal tension according to the frequency in all groups except the 4 week-old group. The % decrease of the maximal developed tension of 4 week-old group at 1 Hz to that of 0.1 Hz after $Na^+$ and $Ca^{2+}$ depletion was only a half of those of the yonger groups. Taken together, it is concluded that the $Na^+-Ca^{2+}$ exchange transports more $Ca^{2+}$ released from SR out of the cell in proportion to the frequency, and this is responsible for the negative staircase effect of the rat heart.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.6
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• pp.725-732
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• 1998

In order to elucidate the molecular mechanism of the intracellular $Ca^{2+}$ overload frequently reported from diabetic heart, diabetic rats were induced by the administration of streptozotocin, the membrane vesicles of junctional SR (heavy SR, HSR) were isolated from the ventricular myocytes, and SR $Ca^{2+}$ uptake and SR $Ca^{2+}$ release were measured. The activity of SR $Ca^{2+}-ATPase$ was $562{\pm}14$ nmol/min/mg protein in control heart. The activity was decreased to $413{\pm}30$ nmol/min/mg protein in diabetic heart and it was partially recovered to $485{\pm}18$ nmol/min/mg protein in insulin-treated diabetic heart. A similar pattern was observed in SR $^{45}Ca^{2+}$ uptakes; the specific uptake was the highest in control heart and it was the lowest in diabetic heart. In SR $^{45}Ca^{2+}$ release experiment, the highest release, 45% of SR $^{45}Ca^{2+}$, was observed in control heart. The release of diabetic heart was 20% and it was 30% in insulin-treated diabetic heart. Our results showed that the activities of both SR $Ca^{2+}-ATPase$ and SR $Ca^{2+}$ release channel were decreased in diabetic heart. In order to evaluate how these two factors contribute to SR $Ca^{2+}$ storage, the activity of SR $Ca^{2+}-ATPase$ was measured in the uncoupled leaky vesicles. The uncoupling effect which is able to increase the activity of SR $Ca^{2+}-ATPase$ was observed in control heart; however, no significant increments of SR $Ca^{2+}-ATPase$ activities were measured in both diabetic and insulin-treated diabetic rats. These results represent that the $Ca^{2+}$ storage in SR is significantly depressed and, therefore, $Ca^{2+}-sequestering$ activity of SR may be also depressed in diabetic heart.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.113-120
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• 1990

Influences of trigger calcium on myocardial contraction from several sources were investigated on the frequency reduction-induced changes of contraction in rat left atria driven by electrical field stimulation. Rat atria elicited characteristic three phase-changes according to frequency reduction: the first rapid rise in twitch tension, the second transient fast decrease in tension and the third maintenance of twitch tension at about 200% of resting tension during high frequency. Caffeine treatment enormously suppressed the frequency reduction-induced twitch tension increase. The atrial contraction during high frequency vanished after verapamil treatment. But, during low frequency, atrial contraction revived in the presence of verapamil. Ouabain treatment and sodium depletion in superfusing solution abolished the characteristic second phase with slow frequency. These results suggest that slow calcium channel is an indispensable calcium entry route and calcium release from sarcoplasmic reticulum is an major source for trigger calcium in cardiac contraction. And sodium-calcium exchange has a modulatory roles in the regualtion of trigger calcium according to the changes of intracellular sodium concentration.

• Applied Microscopy
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• v.20 no.2
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• pp.127-159
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• 1990

This study was investigated to elucidate the effect of chlorambucil on the heart in aging rats. Male rat ranging in age from 3 to 36 months were used. Each age groups(3, 6, 12, 18, 24, 36 months) included control and chlorambucil treated rats. As a part of the study, the ultrastructural changes in the left ventricular myocardial cells were described by using the qualitative and morphometric techniques. The results are summarized as follows. Age associated ultrastructural changes included: an increase in vacuoles, protrusion of plasma membranes, lipid droplets, and lipofuscins in myocardium of control groups. These changes which begin to occur at 12 months and continue through 36 months. At the 36 months some instance of unusual formation of contraction band and separation of intercalated discs were encountered. Morever, these changes and contents with chlorambucil treatment were remarkably increased in comparison with control groups. Age-dependent changes of control group measured with morphometry were not observed in the volume densities of mitochondria and myofibrils. But there was increase in interstitium. On the other hand decrease in sarcoplasmic reticulum and T-tubule system. In chlorambucil treated groups, volume densities of mitochondria and interstitium were increased in comparison with those of the control groups. But sarcoplasmic reticulum and T-tubule system were remarkably decreased.

• The Korean Journal of Physiology and Pharmacology
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• v.6 no.2
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• pp.101-107
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• 2002

In the present study, the postnatal developmental changes in the expressional levels of cardiac sarcoplasmic reticulum (SR) $Ca^{2+}$ regulatory proteins, i.e. $Ca^{2+}-ATPase,$ phospholamban, and $Ca^{2+}$ release channel, were investigated. Both SR $Ca^{2+}-ATPase$ and phospholamban mRNA levels were about 35% of adult levels at birth and gradually increased to adult levels. Protein levels of both SR $Ca^{2+}-ATPase$ and phospholamban, which were measured by quantitative immunoblotting, were closely correlated with the mRNA levels. The initial rates of $Ca^{2+}$ uptake at birth were about 40% of adult rates and also increased gradually during the myocardial development. Consequently, the relative phospholamban/$Ca^{2+}-ATPase$ ratio was 1 in developmental hearts. $Ca^{2+}$ release channel (ryanodine receptor) mRNA was about $50{\sim}60%$ at birth and increased gradually to adult level throughout the postnatal rat heart development. $^3[H]ryanodine$ binding increased gradually during postnatal myocardial development, which was closely correlated with ryanodine mRNA expression levels during the development except the ryanodine mRNA level at birth. These findings indicate that cardiac SR $Ca^{2+}-ATPase,$ phospholamban, and $Ca^{2+}$ release channel are expressed coordinately, which may be necessary for intracellular $Ca^{2+}$ regulation during the rat heart development.