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Biomedical subjects

M Sakakibara

Publications and source records attributed to M Sakakibara.

At least 127 records · Page 7Linked to original sources

Modulation of calcium-mediated inactivation of ionic currents by Ca2+/calmodulin-dependent protein kinase II.

Iontophoretic injection of Ca2+ causes reduction of I0A (an early rapidly activating and inactivating K+ current) and I0C (a late Ca2+-dependent K+ current) measured across the isolated type B soma membrane (Alkon et al., 1984, 1985; Alkon and Sakakibara, 1984, 1985). Similarly, voltage-clamp conditions which cause elevation of [Ca2+]i are followed by reduction of I0A and I0C lasting 1-3 min. Iontophoretic injection of highly purified Ca2+/CaM-dependent protein kinase II (CaM kinase II) isolated from brain tissue (Goldenring et al., 1983) enhanced and prolonged this Ca2+-mediated reduction of I0A and I0C. ICa2+, a voltage-dependent Ca2+ current, also showed some persistent reduction under these conditions. Iontophoretic injection of heat-inactivated enzyme had no effect. Agents that inhibit or block Ca2+/CaM-dependent phosphorylation produced increased I0A and I0C amplitudes and prevented the effects of CaM kinase II injection. The results reported here and in other studies implicate Ca2+-stimulated phosphorylation in the regulation of type B soma ionic currents.

Animals↗

Inositol trisphosphate regulation of photoreceptor membrane currents.

In previous studies elevation of intracellular Ca2+ was shown to cause prolonged reduction of two voltage-dependent K+ currents (IA and ICa2+-K+) across the membrane of the isolated Hermissenda photoreceptor, the type B cell (Alkon et al., 1982b; Alkon and Sakakibara, 1985). Here we show that iontophoretic injection of inositol trisphosphate (IP3), but not inositol monophosphate, also caused prolonged reduction of IA and ICa2+-K+. IP3 injection also caused reduction of a light-induced K+ current (also ICa2+-K+) but did not affect the voltage-dependent Ca2+ current, ICa2+, or the light-induced inward current, INa+, of the type B cell. IP3 injection caused similar effects on the K+ currents of the other type of Hermissenda photoreceptor, the type A cell. INA+ of the type A cell, unlike that of the type B cell, was, however, markedly increased following IP3 injection. The differences of IP3 effects on the two types of photoreceptors may be related to differences in regulation of ionic currents by endogenous IP3 as reflected by clear differences (before injection) in the magnitude of IA, ICa2+-K+, and INa+ between the two cell types.

Animals↗

[A resected case of pleomorphic carcinoma of pancreatic head].

A 41-year-old man was admitted to the hospital complaining of back pain and progressive jaundice. Ultrasonography, CT and hypotonic duodenography revealed a large and well-defined tumor in the head of pancreas. The tumor was hypervascular on angiography. Total pancreatectomy was performed, and the examination of the resected specimen disclosed that the tumor was 50 X 45 mm in size and extrapancreatic tumor-forming type. Histopathologically, the tumor comprized mostly bizzare mono- and multinucleated giant cells with sarcomatous growth pattern. The patient died 6 months after operation due to liver metastasis and peritoneal dissemination. The pathological feature of the pleomophic carcinoma of the pancreas is well-known, but the clinical feature is vague because the resected cases are rare. The pathological finding of the specimen of this case is typical and it is considered that the clinical findings of this case, such as well-defined and hypervascular tumor, are characteristic of the pleomorphic carcinoma of the pancreas.

Adult↗

Nucleotide sequence of a cDNA clone for human aldolase: a messenger RNA in the liver.

Nearly complete cDNA clones for human aldolase A mRNA were isolated from human liver cDNA library and the nucleotide sequence determined. Using the cDNA clone as a probe the length of human aldolase A mRNAs, isolated from the skeletal muscle, liver and placenta tissues, was measured by RNA blotting and estimated to be 1,600 nucleotides for skeletal muscle mRNA and 1,700 nucleotides for both the liver and placenta mRNAs, indicating that different species of mRNA coding for human aldolase A were expressed in the different tissues.

Amino Acid Sequence↗

Human aldolase isozyme gene: the structure of multispecies aldolase B mRNAs.

A complete nucleotide sequence of human aldolase B mRNA was determined with a recombinant cDNA (pHABL120-3). The cDNA insert was composed of 1,652 bases excluding poly(A) tail and the sequence was consistent with the previous results reported by others. However, S1 nuclease mapping and subsequent genomic analysis allowed us to know that the clone possesses two more sites corresponding to 5'-termini in the 5'-noncoding region and another site of polyadenylation in the 3'-noncoding region. In fact, the major aldolase B mRNA species occupying 90% of the total mRNAs initiated at the predominant position corresponding to the position around -82 of the 5'-noncoding sequence in pHABL120-3 and terminated at the distal polyadenylation site. Second species accounting for 9% of the mRNAs initiated at the same site and terminated at the proximal polyadenylation site. The remainings have a longer 5'-noncoding sequence which starts from further upstream region of the major one and pHABL120-3 corresponds to one of these largest clones.

Amino Acid Sequence↗

Calcium activates and inactivates a photoreceptor soma potassium current.

Light-induced currents were measured with a two-microelectrode voltage clamp of type B photoreceptor somata, which had been isolated by axotomy from all synaptic interactions as well as from all membranes capable of generating impulse activity. In artificial seawater (ASW), light elicited a transient early inward current, INa+, which depended on Na+o and had a linear current-voltage relation and an extrapolated reversal potential of 30-40 mV (absolute). In 0-Na+ ASW, light elicited a transient short-latency outward current that dependent on K+o, increased exponentially with more positive voltages (greater than or equal to -40 mV), and reversed at -70 to -75 mV. This outward current was not blocked by Ca++ channel blockers (e.g., Cd++, Co++) or substitution of Ba++o, for Ca++o, but was reduced by iontophoretic injection of EGTA. In both ASW and 0-Na+ ASW, light also elicited a delayed, apparently inward current, which was associated with a decreased conductance, depended on K+o, increased exponentially with more positive voltages (greater than or equal to -40 mV), reversed at the equilibrium potential for K+ flux in elevated K+o was eliminated by substitution of Ba++o for Ca++o, and was greatly reduced by Cd++o or Co++o. Thus, light elicited an early Ca++-dependent K+ current, IC, and a prolonged decrease of IC. Iontophoretic injection of Ca++ through a third microelectrode caused prolonged reduction of both IC and the light-induced decrease of IC, but did not alter ICa++ or the current-voltage relation of IC. Ruthenium red (1 microM) in the external medium caused a prolongation of the light-induced decrease of IC. Iontophoretic injection of EGTA often eliminated the light-induced IC decrease while decreasing peak IC (during depolarizing steps to -5 or 0 mV) by less than one-half. EGTA injection, on the average, did not affect steady state IC but reduced the light-induced decrease of steady state IC to approximately one-third of its original magnitude. The prolonged IC decrease, elicited by dim light in the absence of light-induced IC or INa+, was more completely eliminated by EGTA injection. It was concluded that light, in addition to inducing a transient inward Na+ current, causes both a transient increase and a prolonged decrease of IC via elevation of Ca++i.

Animals↗

Reduction of two voltage-dependent K+ currents mediates retention of a learned association.

A single identified neuron, the medial type B photoreceptor, was isolated by axotomy from the nervous systems of nudibranch molluscs (Hermissenda) which had been exposed to three different training experiences. Paired animals had been trained with repeated paired presentations of light and rotation and random animals with randomized light and rotation; naive animals had no training. A two-microelectrode voltage clamp of axotomized type B somata (separated from all synaptic interactions and impulse activity) was used to measure, with a blind procedure, three distinct ionic currents at least 24 h after the training experience. An early K+ current, IA, and a Ca2+-dependent K+ current, ICa2+-K+, but not a light-induced inward Na+ current, were significantly reduced for the paired as compared to the random and naive animals. The magnitude of ICa2+-K+ reduction was related (again measured blindly) to the degree of training-induced suppression of phototaxis (a measure of the learned behavior) for the paired animals. These data are consistent with previous observations indicating that changes of intrinsic type B membrane properties are an important means for encoding the acquisition and retention of Hermissenda associative learning.

Animals↗

[Magnetic resonance imaging of cardiovascular thrombi].

Magnetic resonance imaging (MRI) was performed for 10 patients with cardiovascular thrombi using a 0.1-Tesla resistive type apparatus (ASAHI MARK-J). In all cases thrombi were clearly imaged by NMR and their shapes closely resembled those imaged by X-ray CT. Mural thrombi located within left ventricular aneurysms of two patients with old anteroseptal myocardial infarction were semilunar in form. A mural thrombus in the right ventricle of a patient with cardiovascular Behcet's disease was also clearly imaged. Mural thrombi within the enlarged left atrium of two patients with mitral valve stenosis and insufficiency were clearly demonstrated in both cross- and longitudinal sections. In three patients with thoracic aortic aneurysm, mural thrombi were recognized within the local dilatations of the aorta. In two patients with dissecting aortic aneurysm, mural thrombi were visualized within false lumen using MRI. Mean T1 values and standard deviations for the left ventricular cavity, the left ventricular wall, and the thrombi were 639 +/- 49, 349 +/- 17 and 316 +/- 84 msec, respectively. Mean T1 values of the thrombi were usually shorter than those of the left ventricular wall. Some supposedly fresh thrombi had longer T1 values, however.

Adult↗

[Magnetic resonance imaging of cardiovascular diseases: advantages of coronal and sagittal planes].

The usefulness of coronal and sagittal sections of the cardiovascular system by magnetic resonance imaging was evaluated. Coronal, sagittal and transverse spin echo scans using ECG-non-gating and gating during systole and diastole were performed for five normal volunteers, 91 with heart diseases (25 valvular disease, 28 ischemic heart disease, 14 cardiomyopathies, 14 congenital malformations, four pericardial diseases, and six others) and 32 patients with aortic abnormalities (17 aneurysms, 10 dissections and five others) using a 2.5 KGauss unit. Cardiac gating necessitated six to eight min per scan, but it was mandatory to obtain clear images of the details. On the other hand, in most of the aortic abnormalities, diagnostic images were obtained by the ECG-non-gating technique which required only about 2.5 min per scan. Coronal and sagittal sections were useful for estimating the entire shape and size of each cardiac chamber and intracardiac thrombi, the extent of postinfarctional wall thinning and cardiac aneurysms, and hypertrophy or narrowing of both the ventricular outflow tracts and apex. These planes were particularly useful, and more contributory than transverse planes for detecting inferior myocardial damage such as infarction. A few coronal and sagittal scans were sufficient to diagnose extensive lesions of the aorta, such as atherosclerosis, dissections and the aortitis syndrome. Local lesions such as coarctation, supravalvular aortic stenosis, annulo-aortic ectasia and aneurysm, especially those originating in the inferior wall of the aortic arch were easily discovered. Since the main arteries, such as the innominate, left common carotid, left subclavian and renal arteries, were clearly demonstrated by coronal images, coronal scans were considered more useful than transverse ones for observing the relationship between these arteries and dissections or aneurysms of the arch and of the abdominal aorta.

Coronary Disease↗

[Aldolase isozyme gene: on the structure and the tissue-specific expression of a muscle type aldolase gene].

A muscle-type aldolase gene known to be a tumor marker enzyme was analyzed. It was found that three different molecular species of the mRNA exist and are expressed in a tissue-specific manner. These mRNAs have identical coding and 3' noncoding sequences and differ only at the 5' end of the sequence. Genomic DNA analysis indicated that a single aldolase gene for one muscle type specifies three different mRNAs by organizing as a leader sequence a region corresponding to each distinct 5' end of the mRNA followed by a shared common structural gene in the genome.

Animals↗

Different expression of rat aldolase A mRNA in the skeletal muscle and ascites hepatoma cells.

Rat aldolase A mRNAs from the skeletal muscle and ascites hepatoma cells are compared by RNA blot hybridization. The hepatoma AH60C and AH130 mRNA are about 1600 bases long, some 50 bases longer than the skeletal muscle transcript. Aldolase A enzymes encoded by these mRNAs, however, are indistinguishable in electrophoresis on cellulose polyacetate strips and two-dimensional polyacrylamide gel. In addition, comparison of the thermal stability of hybrids between the mRNAs from the skeletal muscle and hepatoma cells and a cDNA probe (pRAAM83) made from the aldolase A mRNA of the skeletal muscle indicate that the mRNAs have extremely high sequence homology, at least, within the most part of coding region in addition to the 3' noncoding region. These findings indicate that these mRNA may be transcribed by the use of different promoter sites and the common structural gene.

Animals↗

Voltage-dependent calcium and calcium-activated potassium currents of a molluscan photoreceptor.

Two-microelectrode voltage clamp studies were performed on the somata of Hermissenda Type B photoreceptors that had been isolated by axotomy from all synaptic interaction as well as any impulse-generating (i.e., active) membrane. In the presence of 2-10 mM 4-aminopyridine (4-AP) and 100 mM tetraethylammonium ion (TEA), which eliminated two previously described voltage-dependent potassium currents (IA and the delayed rectifier), a voltage-dependent outward current was apparent in the steady state responses to command voltage steps more positive than -40 mV (absolute). This current increased with increasing external Ca++. The magnitude of the outward current decreased and an inward current became apparent following EGTA injection. Substitution of external Ba++ for Ca++ also made the inward current more apparent. This inward current, which was almost eliminated after being exposed for approximately 5 min to a solution in which external Ca++ was replaced with Cd++, was maximally activated at approximately 0 mV. Elevation of external potassium allowed the calcium (ICa++) and calcium-dependent K+ (IC) currents to be substantially separated. Command pulses to 0 mV elicited maximal ICa++ but no IC because no K+ currents flowed at their new reversal potential (0 mV) in 300 mM K+. At a holding potential of -60 mV, which was now more negative than the potassium equilibrium potential, EK+, in 300 mM K+, IC appeared as an inward tail current after positive command steps. The voltage dependence of ICa++ was demonstrated with positive steps in 100 mM Ba++, 4-AP, and TEA. Other data indicated that in 10 mM Ca++, IC underwent pronounced and prolonged inactivation whereas ICa++ did not. When the photoreceptor was stimulated with a light step (with the membrane potential held at -60 mV), there was also a prolonged inactivation of IC. In elevated external Ca++, ICa++ also showed similar inactivation. These data suggest that IC may undergo prolonged inactivation due to a direct effect of elevated intracellular Ca++, as was previously shown for a voltage-dependent potassium current, IA. These results are discussed in relation to the production of training-induced changes of membrane currents on retention days of associative learning.

Action Potentials↗