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R A Fluck

Publications and source records attributed to R A Fluck.

14 recordsLinked to original sources
PubMed1998

Gravity influences the position of the dorsoventral axis in medaka fish embryos (Oryzias latipes).

To determine whether gravity influences the plane of bilateral symmetry in medaka embryos, zygotes were placed with their animal-vegetal axis orientated vertically and with their vegetal pole elevated. Then, at regular intervals during the first cell cycle, the zygotes were tilted 90 degrees for about 10 min and subsequently returned to their original orientation. In embryos tilted during the first half of the first cell cycle, the embryonic shield formed on the side that had been lowermost when the zygote was tilted. In embryos that were tilted twice, first in one direction and then in the opposite direction, the embryonic shield formed on the side that was lowermost the first time. When zygotes were centrifuged at 5 g, the embryonic shield formed on the outwardly radial (centrifugal) side of the embryo. The orientation of the array of parallel microtubules in the vegetal pole region was also influenced by tilting or centrifuging zygotes. No correlation was found between the positions of the polar body and the micropyle and the plane of bilateral symmetry. It was concluded that gravity influences both the plane of bilateral symmetry and the orientation of microtubules in the vegetal pole region of medaka embryos.

Animals↗
PubMed1993

Calcium buffer injections inhibit cytokinesis in Xenopus eggs.

A slow cortical wave of high calcium accompanies the elongation of cleavage furrows in medaka fish eggs as well as in Xenopus eggs. We explored the role of such waves by injecting calcium buffers into Xenopus eggs at various times before and during first and second cleavage. Injection earlier than about 15 minutes before first cleavage normally starts delays it for hours. Injection between about 15 minutes and a few minutes before cleavage normally starts allows a (short) furrow to form on time but usually yields an eccentric one. This forms away from the injection side, often as far off-center as the egg's equator, and then regresses. Injection soon after it starts quickly arrests elongation of the furrow and eventually induces its regression; while injection a bit later likewise soon arrests elongation but allows delocalized furrow deepening to continue. The dependence of these inhibitory actions upon the dissociation constants and final cytosolic concentrations of the injected buffers indicates that they act as shuttle buffers to suppress needed zones of high calcium in the micromolar range. We conclude that the high calcium that is found within these furrows is needed to induce them, to extend them and even to maintain them. Moreover, while short, eccentric furrows often form as far off center as the equator, they somehow always form along a meridian through the animal pole. This seems difficult to explain by the orthodox, diastral model. Rather, it suggests that the cleavage furrows in Xenopus--and perhaps in animal cells quite generally--are directly induced by a diastema or telophase disc rather than by the asters.

Animals↗
PubMed1991

Slow calcium waves accompany cytokinesis in medaka fish eggs.

Animal cells are cleaved by the formation and contraction of an extremely thin actomyosin band. In most cases this contractile band seems to form synchronously around the whole equator of the cleaving cell; however in giant cells it first forms near the mitotic apparatus and then slowly grows outwards over the cell. We studied the relationship of calcium to such contractile band growth using aequorin injected medaka fish eggs: we see two successive waves of faint luminescence moving along each of the first three cleavage furrows at approximately 0.5 micron/s. The first, narrower waves accompany furrow extension, while the second, broader ones, accompany the subsequent apposition or slow zipping together of the separating cells. If the first waves travel within the assembling contractile band, they would indicate local increases of free calcium to concentrations of about five to eight micromolar. This is the first report to visualize high free calcium within cleavage furrows. Moreover, this is also the first report to visualize slow (0.3-1.0 micron/s) as opposed to fast (10-100 microns/s) calcium waves. We suggest that these first waves are needed for furrow growth; that in part they further furrow growth by speeding actomyosin filament shortening, while such shortening in turn acts to mechanically release calcium and thus propagates these waves as well as furrow growth. We also suggest that the second waves act to induce the exocytosis which provides new furrow membrane.

Animals↗
PubMed1988

Electrical currents associated with rhythmic contractions of the blastoderm of the medaka, Oryzias latipes.

1. We used a vibrating probe to measure extracellular electrical currents near the surface of dechorionated Oryzias latipes eggs as contraction waves moved slowly across the blastoderm. 2. Although we found no detectable current outside dechorionated embryos, we recorded large current pulses near the edge of wounds made in the surface of the blastoderm. 3. The maximum net inward current--or in some cases, the least net outward current--correlated temporally with the contraction of cells near the edge of the wound. 4. The current pulses were superimposed on steady currents of variable magnitude and polarity. 5. We discuss possible mechanisms for the initiation and propagation of the contraction wave.

Animals↗
PubMed1987

Pacemaker region in a rhythmically contracting embryonic epithelium, the enveloping layer of Oryzias latipes, a teleost.

The primary objectives of this study were to determine the embryonic stage at which the Oryzias latipes enveloping layer (EVL) begins to contract rhythmically, and to determine where these contractions arise within the EVL. Using time-lapse video recording, we showed that the contractions begin at stage 14 (the stage of the embryonic shield) and arise in the ventral region of the EVL, which is centered at 180 degrees longitude from the embryonic shield. We have called this the pacemaker region for the contractions. Using fluorescein diacetate as a vital stain, we showed that the ventral region of the EVL continues to act as a pacemaker even after the EVL is detached from the rest of the egg. Rhythmic contractile activity ceased when we removed a group of about 130 cells--10% of the total EVL--from the pacemaker region; comparably large wounds elsewhere had no effect on the contractions. When we cut detached EVLs into ten pieces, only 2.4 +/- 1.8 (mean +/- SD, N = 11) of them contracted rhythmically, even though a considerably larger proportion of the EVL cells participate in the contractions in undisturbed blastoderms. We conclude that the pacemaker cells are necessary for rhythmic contractile activity and that cells outside this region do not contract spontaneously. The contractile waves are propagated at a velocity of 14-54 microns sec-1. This value, which is two to three orders of magnitude slower than the propagation of epithelial action potentials, is similar to the rate of propagation of waves of increased cytosolic Ca2+ in other systems. We propose that the medaka EVL is a good system in which to study certain aspects of epithelial morphogenesis.

Animals↗
PubMed1975

Cholinesterases from plant tissues. VI. Preliminary characterization of enzymes from Solanum melongena L. and Zea mays L.

Enzymes capable of hydrolyzing esters of thiocholine have been assayed in extracts of Solanum melongena L. (eggplant) and Zea Mays L. (corn). The enzymes from both species are inhibited by the anti-cholinesterases neostigmine, physostigmine, and 284c51 and by AMO-1618, a plant growth retardant and they both have pH optima near pH 8.0. The enzyme from eggplant is maximally active at a substrate concentration of 0.15 mM acetylthiocholine and is inhibited at higher substrate concentrations. On the basis of this last property, the magnitude of inhibition by the various inhibitors, and the substrate specificity, we conclude that the enzyme from eggplant, but not that from corn, is a cholinesterase.

Acetylthiocholine↗
PubMed1974

Cholinesterases from Plant Tissues: III. Distribution and Subcellular Localization in Phaseolus aureus Roxb.

The distribution and localization of cholinesterase in Phaseolus aureus, Glycine max, and Pisum sativum is described. The enzyme is present in roots, leaves, stems, root callus tissue, root cells suspension cultures, and root nodules. Cholinesterase in roots is found primarily in the cell wall. In cell fractionation experiments, at least 95% of the cholinesterase activity is associated with cell wall material. The enzyme can be solubilized by salt solutions, whereas Triton X-100 and sodium deoxycholate solubilize relatively small amounts of the enzyme. Cytochemical techniques have been employed to show the presence of cholinesterase activity at the cell surface and in the cell wall of certain cells of the root.

Journal Article↗
PubMed1974

Cholinesterases from plant tissue: v. Cholinesterase is not pectin esterase.

Several properties of the cholinesterase from Phaseolus aureus Roxb. and of pectin (methyl) esterases from both Phaseolus aureus and Lycopersicon esculentum (L.) Mill. are contrasted. Cholinesterase activity is inhibited by all of the concentrations of NaCl tested, from 0.05 m to 0.9 m, a property which differs sharply from published data pertaining to pectin esterase. Although crude preparations of cholinesterase contain pectin esterase activity, further purification by gel filtration of the cholinesterase results in a nearly complete elimination of the pectin esterase activity. The activity of neither the pectin esterase from Lycopersicon esculentum nor that from Phaseolus aureus is affected by 25 mum neostigmine, a potent inhibitor of the cholinesterase activity extracted from Phaseolus aureus.

Journal Article↗