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S Terakawa

Publications and source records attributed to S Terakawa.

At least 19 recordsLinked to original sources
PubMed1991

Exocytosis in colonic goblet cells visualized by video-enhanced light microscopy.

In order to develop a method to quantify the mucus secretion, we observed the mucus epithelium of the rabbit colon under a video-enhanced differential interference contrast microscope. Upon stimulation with muscarinic agonists, secretory granules in individual goblet cells were found to undergo a rapid light intensity change. Simultaneously, the lumen was widened and filled with a cloudy material. In each cell, many of such responses were followed by formation of a large cavity which could be recovered after removal of the stimulant. We infer that the light intensity change of a granule arises from exocytosis. Direct counting of the frequency of these quantal responses would be very useful to monitor the secretory activity of single cell in real time at a high sensitivity.

Animals↗
PubMed1991

Quantitative analysis of exocytosis directly visualized in living chromaffin cells.

Chromaffin cells isolated from the bovine adrenal medulla were observed under a Nomarski microscope through a CCD camera and an image processor. Exocytotic events of individual granules including fusion, extrusion, swelling, omega-figure formation, and membrane retrieval were visualized in individual cells stimulated by acetylcholine or K-rich solution. Initial steps were quicker than 16 ms, and the membrane retrieval was slower than 1-60 s. These findings provided a light microscopic proof for the exocytosis hypothesis as well as a basis for quantification of hormonal release. The technique was used to demonstrate significant secretion induced by a muscarinic agonist.

Acetylcholine↗
PubMed1991

Ionic currents of the nodal membrane underlying the fastest saltatory conduction in myelinated giant nerve fibers of the shrimp Penaeus japonicus.

The myelinated giant nerve fiber of the shrimp, Penaeus japonicus, is known to have the fastest velocity of saltatory impulse conduction among all nerve fibers so far studied, owing to its long distances between nodal regions and large diameter. For a better understanding of the basis of this fast conduction, a medial giant fiber of the ventral nerve cord of the shrimp was isolated, and ionic currents of its presynaptic membrane (a functional node) were examined using the sucrose-gap voltage-clamp method. Inward currents induced by depolarizing voltage pulses had a maximum value of 0.5 microA and a reversal potential of 120 mV. These currents were completely suppressed by tetrodotoxin and greatly prolonged by scorpion toxin, suggesting that they are the Na current. Both activation and inactivation kinetics of the Na current were unusually rapid in comparison with those of vertebrate nodes. According to a rough estimation of the excitable area, the density of Na current reached 500 mA/cm2. In many cases, the late outward currents were induced only by depolarizing pulses larger than 50 mV in amplitude. The slope conductance measured from late currents were mostly smaller than that measured from the Na current, suggesting a low density of K channels in the synaptic membrane. These characteristics are in good harmony with the fact that the presynaptic membrane plays a role as functional node in the fastest impulse conduction of this nerve fiber.

Animals↗
PubMed1991

Exocytosis in living salivary glands: direct visualization by video-enhanced microscopy and confocal laser microscopy.

Although exocytosis is widely believed to involve granule movement, membrane fusion and the emptying of granule content, direct study of these processes has been difficult in living cells because of the limited resolution of conventional light microscopy. Using video-enhanced microscopy and confocal laser microscopy, we have now studied these processes in living rat parotid and submandibular gland acinar cells. Under a differential interference contrast (DIC) microscope equipped with a CCD camera and a high speed image processor, secretory granules were in general stationary even after secretory stimulation with isoproterenol (IPR). Following IPR stimulation, however, there were abrupt changes in light intensity of secretory granules, and many granules disappeared. Confocal microscopy was then performed to confirm whether the observed changes in granules were related to membrane fusion and content release. For this, cells were perfused with the fluid-phase tracer Lucifer Yellow; confocal images thus obtained clearly demonstrated the appearance of fluorescence in omega-shaped invaginations of the apical plasma membrane which corresponded to the sites at which changes were observed in DIC images. The time sequence analyses of confocal images showed that there was a repetitive appearance and disappearance of omega-shaped fluorescent foci at the apical plasma membrane until most of the granules were depleted. During this time, there did not appear to be any significant expansion of the apical plasma membrane and if endocytic uptake of the tracer occurred, it was below the limit of detection. These observations provide new insights into the exocytotic process in salivary glands and are at variance in some respects with previous interpretations made from electron microscopy.

Animals↗
PubMed1989

Lack of effect of a neurotoxin from the scorpion Buthus martensi Karsch on nerve fibers of this scorpion.

A neurotoxin (BmK I) was purified from the venom of the scorpion species Buthus martensi Karsch. Effects of this toxin on the excitability of the abdominal nerve fibers of the same scorpion were examined. The toxin had no effect at all on the action and resting potentials recorded intracellularly even at a concentration as high as 100 microM. A similar result was obtained through optical measurements of the action potential using a potential sensitive dye. Sea anemone toxin II (8 microM) had no effect on nerve excitability either. However, tetrodotoxin (50 nM) reversibly suppressed the action potential and grayanotoxin II (20 microM) induced a sustained depolarization of the nerve membrane which resulted in a reversible suppression of the action potential. BmK I at a concentration of 0.1 microM greatly prolonged the action potential in the crayfish giant axon. We conclude that the Na channel of nerve fibers of this scorpion is totally insensitive to the neurotoxin in this scorpion's venom.

Animals↗
PubMed1989

Turbidity change of the mouse adrenal medulla evoked by acetylcholine stimulation.

The turbidity of the mouse adrenal medulla was measured using a photodiode and a video system equipped with an image processor. Acetylcholine applied at concentrations of 30 to 100 microM induced a slow decrease in turbidity by about 0.1% of the resting value. This response was suppressed by Cd-containing or Ca-deficient medium. These results and a video image suggested a structural change in the chromaffin cells associated with the secretory activity.

Acetylcholine↗
PubMed1988

Purification and the partial amino acid sequence of an insect neurotoxin from the venom of scorpion Buthus martensi Karsch.

1. A neurotoxic peptide was isolated from the venom of the scorpion Buthus martensi Karsch collected in Henan Province, China. 2. This toxin showed the highest neurotoxic potency to crickets amongst all components in the venom examined. 3. The amino acid composition of the toxin was similar to that of insect toxin 1 of Leiurus quinquestriatus quinquestriatus. 4. The partial primary sequence of the toxin at the N-terminal was very similar to that of an insect toxin of Androctonus australis Hector. 5. We conclude that the neurotoxin we isolated is indeed an insect toxin and thus named it as BmK IT.

Amino Acid Sequence↗
PubMed1987

Visualization of secretory activities in the Xenopus neurohypophysis by a high S/N video camera.

The optical turbidity of neurohypophyses of the frog Xenopus laevis was observed with the help of a high signal to noise ratio video camera and a high speed image processor. Electrical stimulation of the pituitary stalk induced a decrease in turbidity of the neurohypophysis. This response was visualized on a monitor screen as a diffuse pattern consisting of many bright spots. The diameters of these spots were similar to those of individual nerve terminals, indicating that the optical response arises from a structural change in individual nerve terminals upon secretory activation.

Animals↗
PubMed1985

Are axoplasmic microtubules necessary for membrane excitation?

The excitability of the squid giant axon was studied as a function of transmembrane hydrostatic pressure differences, the latter being altered by the technique of intracellular perfusion. When a KF solution was used as the internal medium, a pressure difference of about 15 cm water had very little effect on either the membrane potential or excitability. However, within a few minutes after introducing either a KCl-containing, a KBr-containing, or a colchicine-containing solution as the internal medium, with the same pressure difference across the membrane, the axon excitability was suppressed. In these cases, removal of the pressure difference restored the excitability, indicating that the structure of membrane was not irreversibly damaged. Electron-microscopic observations of these axons revealed that the perfusion with a KF solution or colchicine-containing solution preserves the submembranous cytoskeletal layer, whereas perfusion with a KCl or KBr solution dissolves it. These results suggest that the submembranous cytoskeletons including microtubules provide an important mechanical support to the excitable membrane but are not essential elements in channel activities.

Action Potentials↗
PubMed1985

Potential-dependent variations of the intracellular pressure in the intracellularly perfused squid giant axon.

Intracellular pressure responses were recorded from squid giant axons after the axoplasm was removed by the intracellular perfusion technique. A glass tube was inserted into the axon and the movement of the air-water interface formed on the end of the tube was observed with a Y-shaped fibrescope. The intracellular pressure increased and decreased rapidly when an action potential was induced by electrical stimulation. The amplitude of the response was about 10 mPa (or 1 X 10(-3) mmH2O), which was very large in comparison with that observed in unperfused axons. It was sensitive to extracellular Ca2+. The pressure response appeared in an all-or-none manner and could be suppressed by tetrodotoxin. This excluded physicochemical processes on the stimulating electrode or current-supplying electrode as sources of the response. Various other sources of artifacts were also excluded. An extensive removal of the axoplasm by intracellular perfusion with a protease-containing solution and a KCl solution did not eliminate the pressure response. The intracellular pressure was membrane potential dependent, increasing upon depolarization and decreasing upon hyperpolarization of the membrane. Under voltage clamp, the relationship between the membrane potential and the pressure response was parabolic with a maximum at +109 mV (in reference to the resting level). The response did not depend on the membrane current. A much slower response due to electro-osmotic water flow was also detected. The pressure response induced by hyperpolarization of the membrane was suppressed by extracellular application of a lidocaine-containing solution, but not by a tetrodotoxin-containing solution. These results suggest that the pressure responses arise either from a change in electrostriction across the axolemma or from a change in charge-dependent tension along the axolemma.

Action Potentials↗
PubMed1979

Evidence for the utilization of extracellular [gamma-32P]ATP for the phosphorylation of intracellular proteins in the squid giant axon.

Proteins in the squid giant axon were labeled with 32P by in vitro incubation of isolated axoplasm with radioactive [gamma-32P]adenosine triphosphate (ATP) and separated by polyacrylamide sodium dodecyl sulfate gel electrophoresis. The two major phosphorylated regions on the gel had molecular weights of 400,000 and 200,000. These two peaks appear to be neurofilament proteins of squid axoplasm. The same set of proteins was phosphorylated in the axoplasm regardless of whether the [gamma-32P]ATP was applied in situ intracellularly or extracellarly. These results suggest that ATP in the extracellular space is, by some ATP-translocation mechanism, utilized in the process of intracellular phosphorylation. Measurements of the apparent influx of ATP across the squid axon membrane yielded results consistent with the view that ATP in the extracellular fluid could be transported into the axoplasm.

Adenosine Triphosphate↗
PubMed1978

Protein release from the internal surface of the squid giant axon membrane during excitation and potassium depolarization.

The proteins in the perfusate collected from intracellularly perfused squid giant axons were analyzed after being labeled with radioactive 125-I-labeled Bolton-Hunter reagent. The rate of protein release into the perfusate was found to be increased by the following electrophysiological manipulations of the axons: (1) repetitive electrical stimulation at 60 Hz in axons perfused with normal potassium fluoride-containing solution or at 0.125 Hz in axons perfused with tetraethylammonium containing solution, (2) perfusion with 4-aminopyridine solution which induces spontaneous electrical activity in the axon, and (3) depolarization of the axon induced by raising the external potassium concentration. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the proteins released under these conditions yielded molecular weight profiles different from those of the extruded axoplasmic proteins. These observations indicate that there exists, in close association with the axonal membrane, aparticular group of proteins, the solubility of which is readily affected by changes in the state of the membrane.

Action Potentials↗
PubMed1978

Intracellular pH and plateau duration of internally perfused squid giant axons.

The effects of changing the intracellular pH on the action potential duration and other electrophysiological properties were studied in squid giant axons perfused intracellularly with TEA(tetraethylammonium)-containing solutions and under Ca-Na bi-ionic conditions. The duration of the action potential plateau, produced by TEA, was markedly decreased with acidic intracellular solutions and increased with alkaline intracellular solutions. The normalized duration, d, was calculated by dividing the plateau duration by that of the standard intracellular pH of 7.3, and plotted against the intracellular pH. The curve could be expressed by the formula log [d/(D-d)] = n(pH-pK') where D is the saturated value of d, n a constant and pK' the value of pH at which d becomes D/2. The values selected for D and n were 2.6 and 1.4, respectively. The pK' was found to be 7.5. Lowering of the extracellular pH to 6.2 only slightly changed the plateau duration. Voltage clamp analysis revealed that acidic intracellular solutions decreased the size of the inward current and the slope conductance which were measured at the late period of the depolarizing clamping pulse. Alkaline intracellular solutions increased the size of the inward current and the slope conductance measured at this late period. Intracellular perfusion with a low pH solution also shortened the duration of the Ca-Na bi-ionic action potential. It is argued that the remarkable generality of the pH effect on the plateau duration implies the existence of a common mechanism of formation of the plateau under widely differing experimental conditions.

Action Potentials↗
PubMed1976

Alteration of birefringence signals from squid giant axons by intracellular perfusion with protease solution.

The optical signal, arising from a transient birefringence change associated with excitation, was recorded from a squid giant axon together with the membrane potential change, and the effect of removal of the axoplasm on the optical signal was examined. In an unperfused axon, repetitive stimulation at a frequency of about 100 Hz produced two kinds of optical response. The initial response had a brief, spike-like time course and was elicited by each stimulating pulse. The delayed response had a slow time course and the sign of decreased light intensity, and summated with repetitive stimulation. Most of the axoplasm was removed from interior of the axon by intracellular perfusion with solutions containing pronase at a concentration of 0.1 mg/ml. The delayed response could selectively be eliminated by perfusion with a pronase-containing solution for 2-8 min. The result was interpreted as showing that the delayed birefringence signal originates from axoplasm when its gel structure was transiently disturbed by an increased Ca2+ influx associated with excitation. When perfusion was further continued the duration of the action potential started increasing and often a prominent after-depolarization appeared. At this stage the initial optical response was again followed by a large slow signal with the sign of increased light intensity. This reversed delayed response was tentatively assumed to originate from the membrane with some remaining axoplasm, but its cause is still not understood.

Animals↗