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PubMed1977

Anomalous permeabilities of the egg cell membrane of a starfish in K+-Tl+ mixtures.

The electrical properties of "inward" rectifying egg cell membranes of the starfish mediastera aequalis have been studied in the presence of K(+)-Tl(+) mixtures. When the ratio of the external concentrations of these ions is changed while their sum is kept constant, both the conductance and the zero-current membrane potential go through a minimum, showing clear discrepancies from theoretical results based on conventional electrodiffusion models (E.g., Goldman's equation). By contrast, when the ration of the two concentrations is fixed and their sum varied, the potential follows an ideal Nernst slope, consistent with Goldman's equation. The membrane conductance which, according to previous studies on similar membranes, is to be viewed as a function of the displacement of the membrane potential from its resting value deltaV, shows marked differences between the cases in which K(+) or Tl(+) are the predominant ions: when K(+) is the predominant permeant ion in solution, the addition of small amounts of Tl(+) inhibits the current, while corresponding blocking effects of K(+) on the current are not observed when Tl(+) is the predominant permeant ion. Also, the time course of the conductance during voltage clamp is different in the two cases, being much faster in Tl(+) than in K(+) solution for comparable values of deltaV. Most of the above features are accounted for by a model in which it is assumed that the ionic channels have external binding sites for cations and that their permeability properties depend on the species of the cation bound (K(+)or Tl(+) in the present experiments).

Animals↗

PubMed1976

Isolation of myosin from starfish sperm heads.

Myosin was extracted and partially purified from the head portion of spermatozoa of the starfish, Asterias amurensis. The sperm myosin showed a specific Ca2+-activated ATPase [EC 3.6.1.3] activity of 0.2 mumoles Pi/min/mg at high ionic strength and pH 6.5. It resembled egg myosin in forming thick filaments, becoming attached to actin filaments. subunit composition, and serological properties.

Adenosine Triphosphatases↗

PubMed1976

New types of gangliosides in starfish with sialic acid residues in the inner part of their carbohydrate chains.

Three gangliosides were isolated from the starfish by Folch's partition, dialysis, silicic acid column chromatography, Iatrobead column chromatography and preparative thin-layer chromatography. Partial acid hydrolysis of the three lipids in 0.1 N HCl at 80 degrees for 90 min liberated all the lactosyl ceramide and water-soluble degradation products, but no sialic acid. Gas-liquid chromatography of the trimethylsilylated methyl glycosides of the water-soluble products demonstrated the presence of arabinose in addition to the usual sugar units of gangliosides.

Gangliosides↗

PubMed1979

Studies on the glycosphingolipids of the starfish, Asterina pectinifera. III. Isolation and structural studies of two novel gangliosides containing internal sialic acid residues.

Two gangliosides, provisionally named Gangliosides 1 and 2 in previous studies, were isolated from starfish, Asterina pectinifera by silicic acid, DEAE-Sephadex, and Iatrobeads column chromatography procedures, and preparative thin-layer chromatography, and their structures were established. On the basis of the results of partial acid hydrolysis, methylation and oxidation with chromium trioxide, Gangliosides 1 and 2 were proposed to be Ara p beta(1 leads to 6)Gal p beta(1 leads to 4)8-O-MeNeuGc(2 leads to 3)Gal p beta(1 leads to 4)Glc p beta(1 leads to 1)-ceramide and Ara p beta(1 leads to 6)Gal p beta(1 leads to 4)NeuGc(2 leads to 3)Gal p beta(1 leads to 4)Glc p beta(1 leads to 1)-ceramide, respectively. The ceramide moieties of both gangliosides had similar phytosphingosine and 2-hydroxy fatty acid compositions, and both gangliosides were structurally related to the previously described Ganglioside 3.

Animals↗

PubMed1978

Blocking effects of barium and hydrogen ions on the potassium current during anomalous rectification in the starfish egg.

1. The blocking effects of Ba+ and H+ on the inward K current during anomalous rectification of the giant egg membrane of the starfish, Mediaster aequalis, were studied using voltage clamp techniques. 2. External Ba2+ at a low concentration (10--100 micron) suppresses the inward K current; the extent of suppression, expressed as the ratio of currents with and without Ba2+, can be described by a conventional bimolecular adsorption isotherm, K/(K + [Ba2+]o), K being an apparent dissociation constant. 3. The dissociation constant, K, decreases as the membrane potential V becomes more negative and can be expressed by K(V) = K(0) exp (zmuFV/RT), where K(0) is the K at V = 0, z is the charge of the blocking ion, and mu is a parameter for the membrane potential dependence of Ba2+ blockage. The value of mu ranges between 0.64 and 0.68. 4. Upon a sudden change in membrane potential the change in the blocking effect of Ba2+ follows first order kinetics; the forward rate constant is membrane-potential-dependent whereas the backward constant is potential-independent. 5. The blocking effect of Ba2+ appears to be independent of the activation of K channels during anomalous rectification. 6. The blocking effect of Ba2+ depends on V alone, in contrast to the activation of the K channel during anomalous rectification which depends on V--VK. 7. In these respects, the effect of Ba2+ is equivalent to the introduction of inactivation into the anomalous rectification. 8. SI2+ and Ca2+ show small but observable blocking effects only at much higher concentrations (about 10--20 mM). 9. The inward K current is suppressed when the external pH is reduced below 6.0. The blocking effect of H+ shows no significant potential dependence. The concentration dependence suggests that three H+ ions simultaneously titrate the acidic groups of each channel (pK = 5.3--5.4). 10. The implications of these results are discussed in terms of molecular models of the potassium channel of anomalous rectification and possible mechanisms of K channel inactivation.

Animals↗

PubMed1979

Effects of internal potassium and sodium on the anomalous rectification of the starfish egg as examined by internal perfusion.

1. The effects of alterations of the intracellular ionic composition on the properties of anomalous (or inward) rectification of the egg membrane of the starfish, Mediaster aequalis, were studied by using an intracellular perfusion technique. The following results were obtained, analysing the membrane current with the voltage-clamp technique. 2. The inward rectification of the K conductance depends only on the membrane potential, V, when the K equilibrium potential, VK, is altered by changing the internal K+ concentration at a fixed external K+ concentration, while it depends on V-VK when VK is altered by changing the external K+ at a fixed internal K+ concentration. 3. From the above the conclusion is reached that the gating of the K channel of the inward rectification depends on V and external but not internal K+ concentration. 4. The conductance of the K channel at a given voltage is roughly proportional to the square root of [K+]i when the latter is altered at a fixed external K+ concentration. 5. Since the conductance is proportional to the square root of [K+]o when this is altered at a fixed internal K+ concentration, the final conclusion is that this conductance is proportional to the geometric mean of the external and internal K+ concentrations. 6. Intracellular Na+ ions are necessary for the activation of inward rectification; the K conductance increases sharply with internal Na+ concentration, reaching saturation at 200 mM. 7. A similar potentiating effect is found for Li+, although it is weaker. Rb+, Cs+ and organic cations such as arginine+ do not have this effect.

Animals↗

PubMed1978

DNA sequence organization in the common Pacific starfish Pisaster ochraceous.

The sequence arrangement of the genomic DNA from the common sea star Pisaster ochraceous has been examined. Reassociation kinetics at DNA fragment lengths of 300 base pairs (bp) indicate the presence of at least three repetitive components in this DNA. The majority of these repetitive sequences are reiterated over the range from 10 to 100's. Approximately one-third of the nucleotides are found in repetitive sequences. Analysis of the reassociation kinetics of 3000-bp DNA fragments demonstrates the interspersion of repetitive and unique DNA sequences. The hyperchromicity of 3000-bp fragments reassociated to low Cot values (the product of moles of nucleotide per litre and time in seconds), and the size distribution of S1 nuclease resistant DNA duplex in these reassociation products, indicate a short-period interspersion pattern in the starfish genome. Repetitive segments (400 +/- 100 bp) are interspersed with longer unique DNA sequences. At a fragment length of 3000 bp the major fraction of the single-copy DNA is found in such an arrangement. In addition to short repetitive segments a substantial portion of the repetitive DNA nucleotides are found in segments excluded by Sepharose CL-2B (greater than or equal to 2000 bp). As much as one-quarter of the repetitive sequence nucleotides can be assigned to long segments.

Animals↗

PubMed1975

Partial purification and properties of phospholipase A2 from the starfish, Asterina pectinifera.

1. Phospholipase A2 (phosphatide 2-acyl-hydrolase, EC 3.1.1.4) activity was shown to occur in the supernatant fraction of a freshly prepared homogenate from the pyloric caecum of starfish (Asterina pectinifera). 2. The phospholipase A2 has been isolated and purified 130-fold by ultracentrifugation, ammonium sulfate precipitation and column chromatographic procedures. 3. The purified enzyme was stable to heat at low pH values and the optimal pH was observed at approximately 9.0. 4. The enzyme activity was activated by Ca2+ and sodium deoxycholate, and was inhibited by EDTA.

Animals↗

PubMed1979

Relationships between hormone-induced calcium release and 86rubidium uptake stimulation in starfish oocytes.

86Rubidium+ uptake, but not 86Rubidium efflux, is strongly stimulated after addition of the meiosis inducing hormone 1-methyladenine (1-MeAde) to prophase blocked oocytes of the starfish Marthasterias glacialis. This stimulation is a transient process which does not require the continuous presence of 1-MeAde and is elicited within 1 minute of contact. 1-MeAde and its biologically active structural analogs fully stimulate Rb+ uptake at concentrations which are about two orders of magnitude lower than those required to trigger meiosis reinitiation but which already release underthreshold levels of Ca2+ from the inner part of the plasma membrane. External Ca2+ concentrations effective in triggering meiosis reinitiation also stimulate Rb+ influx, while drugs like D600, theophyllin and caffein which suppress the hormone induced Ca2+ release, simultaneously preclude the stimulation of Rb+ uptake. Dithiothreitol (DTT) which mimicks 1-MeAde action in releasing Ca2+ and inducing meiosis acts both on the efflux and on active and passive Rb+ influxes. Ouabain, the classical inhibitor of the Na+, K+ pump does not preclude meiosis reinitiation under the influence of 1-MeAde, its agonists of mimetics. It suppresses the active component of Rb+ uptake both in control or stimulate oocytes. When applied only in preincubation before starting the hormone treatment, it cannot however inhibit the stimulation of Rb+ uptake, while basal pump inhibition is preserved. These results demonstrate that stimulation of the active Rb+ or K+ transport is not indispensable to meiosis reinitiation. They suggest moreover that the hormone induced Ca2+ release from the plasma membrane may be responsible for unmasking new ouabain sensitive transport sites.

Adenine↗

PubMed1978

Studies on dwarf larvae developed from isolated blastomeres of the starfish. Asterina pectinifera.

Not only a whole denuded egg, but also blastomeres isolated from 2-, 4- and 8-cell starfish embryos developed into morphologically normal, but dwarf bipinnariae, the sizes of which were roughly proportionate to that of the respective original blastomeres. Some of the blastomeres isolated from the 16-cell stage were also capable of developing into the larval stage. All isolated blastomeres divided in good synchrony with the control embryos. Blastulae of all groups gastrulated within quite a short range of time, around 14-5 h after insemination at 20+/-1 degrees C, although one-third of the 1/8-blastula missed this chance but gastrulated by 19.5 h. The number of constituent cells of the 1/8-gastrula was counted to be about 560, which corresponds roughly to one-half that of the 1/4-, one-fourth of the 1/2- and one-eight of the 1/1-gastrula. This ratio also fitted roughly for the total cell volume. The results are compared with those of other invertebrate species, as well as of some vertebrates, and are discussed in connexion not only with the concepts of 'regulative' and 'mosaic' eggs, but also with a criterion that does not fit into either of these; the developmental system of the mammals.

Animals↗

PubMed1977