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Biomedical subjects

M Sokabe

Publications and source records attributed to M Sokabe.

At least 55 records · Page 3Linked to original sources
PubMed1994

Enzymatic and genetic adaptation of soleus muscle mitochondria to physical training in rats.

To evaluate the effects of physical training on mitochondrial gene expression and mitochondrial biogenesis in slow-twitch muscle, adult female Sprague-Dawley rats were trained for 3, 6, and 12 wk by running on a motor-driven treadmill (speed of 25 m/min and duration of 90 min/day, 5 days/wk), and the activities of citrate synthase, ubiquinol-cytochrome-c oxidoreductase, cytochrome oxidase, mitochondrial cytochrome b mRNA (by Northern blot analysis), and mitochondrial DNA (by slot-blot and Southern blot analyses) were measured in rat soleus muscle. A DNA probe for detection of mitochondrial mRNA and DNA was prepared from a 1,500-bp fragment of human mitochondrial DNA that included the coding region of the cytochrome b gene. Training for 3, 6, and 12 wk significantly increased the activities of citrate synthase (31, 28, and 47%, respectively), ubiquinol-cytochrome-c oxidoreductase (61, 63, and 77%, respectively), and cytochrome oxidase (25, 26, and 32%, respectively) in muscle. The concentration of cytochrome b mRNA in the muscle was proportionally elevated with the enzyme activities. On the other hand, the mitochondrial DNA concentration in the muscle was not altered by training for 3 or 6 wk but increased significantly after training for 12 wk (35% in the slot-blot analysis and 31% in the Southern blot analysis). These results suggest that an increase in the oxidative capacity of slow-twitch muscle by the relatively short-term training is regulated at the pretranslational step in mitochondrial protein synthesis but that the increase by the long-term training involves mitochondrial replication.

Adaptation, Physiological↗
PubMed1993

Involvement of stretch-activated ion channels in Ca2+ mobilization to mechanical stretch in endothelial cells.

Endothelial cells are subjected to shear stresses by blood flow, normal stresses by blood pressure, and stretch by vessel expansion. These forces are known to induce secretions of several vasoactive substances probably via internal calcium mobilization (R. F. Furchgott. Circ. Res. 53: 557-573, 1983; M. J. Peach, A. L. Loeb, H. A. Singer, and J. Saye. Hypertension Dallas 7, Suppl. I: I-94-I-100, 1985). Here we report that stretching cellular membranes increased intracellular Ca2+ concentration ([Ca2+]i) in human umbilical endothelial cells cultured on silicon membranes. Upon application of a stretch pulse (3-s duration), [Ca2+]i increased rapidly and decayed slowly. The following results suggest that this increase arises from Ca2+ entry through stretch-activated (SA) channels: 1) the Ca2+ response disappeared when extracellular Ca2+ was removed; 2) gadolinium (Gd3+), a blocker for cation-selective SA channels, blocked the response but nifedipine did not; and 3) externally applied Mn2+, which is known to permeate mechanosensitive channels but not Ca2+ channels, entered the intracellular space immediately after an application of mechanical stretch. The increase in [Ca2+]i was found to consist of at least two components: an initial fast component and a delayed slower component. Ryanodine inhibited the slow component. It is suggested that stretching the membrane primarily induced extracellular Ca2+ entry through SA channels followed by Ca2+ releases from intracellular Ca2+ stores.

Calcium↗
PubMed1993

Plateau pattern of afferent discharge rate from frog muscle spindles.

1. A characteristic plateau pattern was observed in the rate of afferent discharges during ramp-and-hold stretch of spindles isolated from semitendinosus muscles of frogs. The plateau pattern was more frequent in summer frogs (84%) than winter frogs (11%). 2. The plateau pattern consisted of a discharge rate around 120 imp/s at the end of dynamic stretch, followed by second and third steps of plateau rates around 60 and 40 imp/s, respectively. The intervals of impulses in lower steps were approximately n times those of the top step. 3. The plateau pattern was not sensitive to cutting extracapsular myelinated branches or lowering temperature from 23 to 12 degrees C. However, the number of the plateau was reduced in both cases. 4. Application of depolarizing current to the sensory terminal abolished the plateau pattern. In contrast, in spindles that did not show a plateau pattern, hyperpolarizing current induced such a pattern. 5. Calcium channel blockers and protein kinase C inhibitors abolished the plateau pattern. The plateau pattern could be established in quiescent spindles by drugs eliciting Ca2+ entry, raising cytosolic-free Ca2+, and activating protein kinase C. 6. The most striking aspect of the present study is the stability in the discharge rate at each step of the plateau, irrespective of different experimental conditions. This suggests that the spindle sensory terminal possesses a stable intrinsic rhythm generator in excitation, of which maximum frequency is 120 imp/s. The generator seems to be triggered by stretch stimulus and to be regulated by cytoplasmic Ca2+ and protein kinase C.

Afferent Pathways↗
PubMed1993

Mechanics of patch clamped and intact cell-membranes in relation to SA channel activation.

Stretch activated (SA) channels are believed to be activated by tension in the membrane generated by membrane stretch. However, very few studies have been made on the quantitative estimation of the tension during membrane stretch. Here we present a method to evaluate the tension both in patch clamped and intact cell-membranes. The tension in patch clamped membranes was calculated from Laplace's law by knowing transmembrane pressure and the radius of patch-curvature. We also provide a simpler version for calculating the tension from the pressure and pipette radius. The tension in intact cell membranes was calculated from Hook's law based on the measurement of changes in cell surface area. The estimated tension required for activating SA channels in both types of membranes was found to be comparable suggesting that the SA channel acts as a physiological mechanotransducer in intact cells.

Animals↗
PubMed1993

[Involvement of stretch activated (SA) ion channels in cardiovascular responses to mechanical stimuli].

Cardiovascular system exhibits various responses to mechanical stimulations. Overload in heart induces arrhythmias or hypertrophy. Increases in blood flow or blood pressure elicit vessel dilation via an activation of endothelial cells. Although the molecular mechanism of these responses are unclear, the evidence has been accumulated that the stretch activated (SA) ion channel acts as a primary sensor for mechanical stimuli. In this article, cardiovascular responses to mechanical stimuli have been summarized and the possible role of SA channels is discussed.

Animals↗
PubMed1992

Mechanotransducing ion channels in astrocytes.

Ion channels present on the soma of neonatal rat astrocytes in primary cell culture were studied using the single channel recording technique. Ion channels were activated by changing the pressure in the back of the pipette. The morphological structure of the patch membrane was examined while recording channel activity. One class of channel was activated by increasing the pipette pressure (curvature-sensitive or CS channels). CS channels were observed in 150 mM KCl, 150 mM NaCl, or 150 mM sodium gluconate. At constant pressure the closed times decreased with depolarization. CS channels had a conductance of 50 pS in 150 mM NaCl, and displayed an inwardly rectifying current-voltage relationship. CS channel activity was found only in cell-attached patches, and were active only when the patch membrane curved towards the soma. The other class of channel was found to be activated by both suction and pressure (stretch-activated or SA channels). Four SA conductance levels were found: 360, 230, 144, and 70 pS in 150 mM KCl. Each conductance displayed a linear current-voltage relationship. At negative membrane potentials SA channels were inhibited by Cs+, Ba2+ or Na+. The relationship between average mechanosensory current and pressure was biphasic for SA channels and monophasic for CS channels. Combinations of SA and CS channels could be observed in the same patch. We propose that CS channels are non-specific cation channels which sense membrane tension only when the patch membrane is in a specific, permissive curvature. SA channels appear to be K(+)-selective channels that sense membrane tension independent of the direction of curvature.

Animals↗
PubMed1992

Hair cell regeneration in the adult budgerigar after kanamycin ototoxicity.

Adult budgerigars were given kanamycin at a dose of 200 mg/kg/day for 10 successive days. At 1, 7, 14 and 28 days after the drug treatment, the cochleae of the birds were processed for scanning electron microscopy (SEM). Complete degeneration of sensory hair cells was observed in the basal 55-75% of the basilar papilla immediately after the treatment. Regenerating hair cells, characterized by clusters of microvilli and small apical surfaces, were present in the basal end of the papilla as early as one day post-treatment. During the 28 day recovery period, the number of hair cells progressively increased beginning at the base and spreading toward the apex. Although the appearance of the basilar papilla had improved considerably by 28 days post-treatment, the sensory epithelium still contained a number of pathologies, most noticeably, incomplete restoration of hair cell number in the most apical part of the damaged region and the disorganization of hair cell packing. These remaining pathologies may be responsible for the permanent threshold shifts observed in budgerigars exposed to the same dose of kanamycin treatment (Hashino and Sokabe, 1989).

Animals↗
PubMed1991

Anion channels from rat brain synaptosomal membranes incorporated into planar bilayers.

Synaptic membranes from rat brain were incorporated into planar lipid bilayers, and the characteristics of two types of anion-selective channels (type I and type II) were investigated. In asymmetric BaCl2 buffers (cis, 100 mM/trans, 25 mM), single channel conductances at -40 mV were 70 pS (type I) and 120 pS (type II). Permeability ratios (PNa:PBa:PCl) calculated from the Goldman-Hodgkin-Katz current equation for type I and type II channels were 0.23:0.04:1 and 0.05:0.03:1, respectively. Both channels exhibited characteristic voltage-dependent bursting activities. Open probability for type I channels had a maximum of approximately 0.7 at about 0 mV and decreased to zero at greater transmembrane potentials of either polarity. Type II channels were relatively voltage independent at negative voltages and were inactivated at highly positive voltages. Type I channels showed spontaneous irreversible inactivation often preceded by sudden transition to subconducting states. DIDS blocked type I channels only from the cis side, while it blocked type II channels from either side.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1991

Hair cell damage and recovery following chronic application of kanamycin in the chick cochlea.

Three-day old chicks were given kanamycin at a dose of 200 mg/kg/day for 10 days and their cochleae were processed for scanning electron microscopy at 1, 3, 7 and 14 days following the last injection. Both hair cells and supporting cells were damaged by kanamycin in the basal 35% of the basilar papilla. By 14 days post-treatment, however, most of the damaged region had been replaced with regenerating hair cells and supporting cells. The base-to-apex gradient of morphological development along the cochlea was observed in the process of regeneration. Kinocilium and microvilli were observed on the apical surfaces of the regenerating hair cells.

Animals↗
PubMed1991

Quantitative video microscopy of patch clamped membranes stress, strain, capacitance, and stretch channel activation.

Membrane patches from chick skeletal muscle were stretched by applying controlled suction or pressure to the pipette. From images of the patch, the patch dimensions (area and radius of curvature) were computed by nonlinear regression of the images to a geometric model. With no applied pressure, patch membranes are nearly planar and normal to the wall of the pipette. With increasing pressure gradients, the patch bulges, the radius of curvature decreases, and the area increases. The patch capacitance changes in exact proportion to the change in area at a rate of 0.7 microF/cm2. The increase in area is due to a flow of lipid (with perhaps small amounts of diffusible protein) along the walls of the pipette into the patch. The flow is reversible with a relaxation of the pressure gradient. The area elastic constant of the membrane is approximately 50 dyn/cm, insensitive to cytochalasin B and probably represents the elasticity of the underlying spectrin/dystrophin network. Simultaneous measurements of stretch activated (SA) ion channel activity in the patch showed that the sensitivity of channels from different patches, although different when calculated as a function of applied pressure, was the same when calculated as a function of tension. Because patch lipid is free to flow, and hence stress-free in the steady state, SA channels must be activated by tension in the cytoskeleton.

Animals↗
PubMed1991

Electrophysiological analysis of structural aspects of voltage-dependent SR K+ channel.

This article presents a brief review on the electrophysiological analysis of the structural aspects of the voltage-dependent SR (sarcoplasmic reticulum) K+ channel. In the first half, early attempts to determine the physical dimensions of the ion conducting mechanism such as the mouth, narrow tunnel, or ion selective filter of the channel, are reviewed. The depicted cartoon of the SR K+ channel, as an extremely short, busy district with a big mouth on each side, is quite similar to the recently-obtained reconstructed structural image of the acetylcholine receptor channel. In the latter half, we introduce our recent attempts to draw a physical image of the gating mechanism of the SR K+ channel. We examined, for example, the location of the gate and the voltage sensor, and the relationship between them. It is suggested that the gate and the sensor are connected tightly and that the sensor would be exposed to the surface of the lumen side of SR when the gate opens. Finally, the issue of substates in SR K+ channel is discussed. It is implied that the substrate-conductances reflect a partial occlusion of the pore by an intermediate-open gate.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1990

Intramembrane particles and responses of sensory axon terminals during reinnervation of the frog muscle spindle.

Changes in the density of intramembrane particles (IMPs) of sensory nerve terminals in the bullfrog muscle spindle were correlated with recovery in the response of the spindle to stretch during postcrush reinnervation. A few IMPs on the protoplasmic (P) face in summer experiments (June to October) reappeared by the 3rd week after the nerve crush, then rapidly increased to 110% and 120% of control values 2 and 2.5 months after the crush. Afferent responses to stretch could be recorded after the mean IMP density on the P-face in terminal branches had recovered to more than 25% of the control value. The discharge rate showed a plateau pattern during the period of the excessive IMPs. This was supplanted by a normal pattern after a myelinated branch of the sensory axon was cut. The IMPs in winter experiments (November to April) reappeared by day 90 after nerve crush, and then slowly increased. The sustained responses to stretch reappeared after 5 months, when the mean IMP density on the P-face was restored to 25% or more of the control. Neither excessive density of the IMPs nor plateau pattern of the discharge rate were observed in winter experiments. The relation between the regenerated IMP densities and the functional recovery is discussed.

Action Potentials↗
PubMed1990

Aminoglycoside blockade of Ca2(+)-activated K+ channel from rat brain synaptosomal membranes incorporated into planar bilayers.

Ca2(+)-activated K+ channels from rat brain synaptosomal membranes were incorporated into planar lipid bilayers, and the effects of aminoglycoside antibiotics on the single channel conductance (258 +/- 13 pS at 100 mM K+) were investigated. Aminoglycosides reduced the single channel conductance from the 'cis' (cytoplasmic) side in a dose- and voltage-dependent manner. Voltage dependence of the blockade indicated an interaction between positively charged amino residues of aminoglycoside antibiotics and a binding site located within the electric field of the ion-conducting pathway. The order of blocking potency was consistent with that of the number of amino residues of aminoglycosides (neomycin (6) greater than dibekacin (5) greater than ribostamycin (4) = kanamycin (4], while the electrical distance (z delta = 0.46-0.49) of the binding site kept almost constant for each drug. These z delta s were almost the same with those (0.46-0.51) of alkyl-diamine blockers with two amino residues (total net charge of +2) and approximately twice of those (0.25-0.26) of alkylmonoamine blockers (total net charge of +1). Assuming that amino residues of aminoglycosides and alkylamines shared the same binding site located at 25% voltage drop from the cytoplasmic surface of the channel, the site would have to be at least large enough to accommodate one diamino sugar residue of the aminoglycoside in order to simultaneously interact with two positively charged amino groups. Dose- and voltage-dependent blockade of the channel by gallamine, an extremely bulky trivalent organic cation, supported the picture that the channel has a wide mouth on the cytoplasmic side and its 'pore' region, where voltage drop occurs, may also be quite wide and nonselective, suddenly tapering to a constriction where most charged cations block the channel by 'occluding' the K(+)-conducting pathway.

Amines↗