scholarly journals Intracellular pH of sea urchin eggs measured by the dimethyloxazolidinedione (DMO) method.

1981 ◽  
Vol 89 (2) ◽  
pp. 284-291 ◽  
Author(s):  
C H Johnson ◽  
D Epel
Keyword(s):  
Intracellular Ph ◽  
Sea Urchin ◽  
General Result ◽  
Strongylocentrotus Purpuratus ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
Cell Divisions ◽  
Ph Microelectrodes

Intracellular pH (pH1) of sea urchin eggs and embryos was determined using DMO (5,5-dimethyl-2,4-oxazolidinedione). By this method, the pH1 of Lytechinus pictus eggs increased after fertilization from 6.86 to 7.27, and this higher pHi was maintained thereafter, as has been previously observed with pH microelectrodes. The same general result was obtained with the eggs of Strongylocentrotus purpuratus, in contrast to previous estimates of the pH of egg homogenates from this species, which had indicated a rise and then fall of pHi after fertilization. pHi did not significantly change during early cell divisions. Studies of treatments that alter pHi confirmed that ammonia alkalizes and acetate acidifies the cells. The regulation of pHi by embryos in the acidic seawater is impaired if sodium is absent, whereas unfertilized eggs can regulate pHi in acidic, sodium-free seawater.

The Respiratory and Glycolytic Enzymes of Sea-Urchin Eggs

1954 ◽  
Vol 31 (2) ◽  
pp. 208-217
Author(s):  
MARTYNAS YČAS
Keyword(s):  
Cytochrome C ◽  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Lactic Dehydrogenase ◽  
Glycolytic Pathway ◽  
Endogenous Respiration ◽  
Centrifugal Field ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg

1. Activity corresponding to phosphoglucomutase, phosphohexoisomerase, aldolase, triosephosphate dehydrogenase, enolase and lactic dehydrogenase has been demonstrated in homogenates prepared from unfertilized sea-urchin eggs (Strongylocentrotus purpuratus and Lytechinus pictus). 2. The presence of cytochromes a and b1 has been confirmed. These cytochromes sediment in a relatively low centrifugal field. 3. No cytochrome c could be demonstrated, although cytochrome c is both reduced and oxidized by homogenates, and addition of cytochrome c increases the endogenous respiration and oxidation of succinate. 4. These results support the view that the usual glycolytic pathway operates in the sea-urchin egg and is the principal route of oxidation of carbohydrate.

The Effects of Spaceflight Conditions on Calcium-Dependent Secretion and Cytoskeletal Organization During Fertilization and Cell Divisions In the Sea Urchin Lytechinus Pictus

1999 ◽  
Vol 5 (S2) ◽  
pp. 1070-1071
Author(s):  
Heide Schatten ◽  
Amitabha Chakrabarti ◽  
Howard Levine ◽  
Mario Runco ◽  
Ken Anderson ◽  
...  
Keyword(s):  
Sea Urchin ◽  
Research Facility ◽  
Cytoskeletal Organization ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
Calcium Dependent ◽  
Kennedy Space Center ◽  
Cell Divisions ◽  
Kennedy Space ◽  
Calcium Loss

Calcium loss and muscle atrophy are two of the main metabolic changes experienced by astronauts and crew members during exposure to microgravity in space. To investigate the effects of spaceflight on calcium-dependent secretion and cytoskeletal formation in a less complex system we utilized sea urchin eggs and embryos which were fertilized and cultured under spaceflight conditions during the STS-77 shuttle mission. Sea urchin eggs were fertilized and cultured in the newly developed aquatic research facility (ARF) which allowed culture of eggs and embryos in microgravity and in a 1g centrifuge in space. This allowed analysis of the comparison of microgravity and 1g spaceflight treatments with samples cultured on ground. Eggs and embryos were maintained in Standard Container Assemblies (SCAs) with identical sets prepared for culture in microgravity, and at 1g in the middeck compartment of the shuttle Endeavor, as well as for ground observations at the Kennedy Space Center.

Intracellular pH and activation of sea urchin eggs after fertilisation

Nature ◽  
1976 ◽  
Vol 262 (5570) ◽  
pp. 661-664 ◽  
Author(s):  
James D. Johnson ◽  
David Epel ◽  
Miles Paul
Keyword(s):  
Intracellular Ph ◽  
Sea Urchin ◽  
Sea Urchin Eggs

Metabolic importance of Na+/K+-ATPase activity during sea urchin development.

1997 ◽  
Vol 200 (22) ◽  
pp. 2881-2892 ◽  
Author(s):  
P Leong ◽  
D Manahan
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Sea Urchin ◽  
Sodium Pump ◽  
Sea Water ◽  
Strongylocentrotus Purpuratus ◽  
Lytechinus Pictus ◽  
Stimulation Of

Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+-ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+-ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development.

Autonomous and non-autonomous differentiation of ectoderm in different sea urchin species

Development ◽  
1995 ◽  
Vol 121 (5) ◽  
pp. 1497-1505 ◽  
Author(s):  
A.H. Wikramanayake ◽  
B.P. Brandhorst ◽  
W.H. Klein
Keyword(s):  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Protein A ◽  
Cellular Interactions ◽  
V Protein ◽  
Lytechinus Pictus ◽  
Sea Urchin Embryo ◽  
Oral Ectoderm ◽  
Temporal And Spatial

During early embryogenesis, the highly regulative sea urchin embryo relies extensively on cell-cell interactions for cellular specification. Here, the role of cellular interactions in the temporal and spatial expression of markers for oral and aboral ectoderm in Strongylocentrotus purpuratus and Lytechinus pictus was investigated. When pairs of mesomeres or animal caps, which are fated to give rise to ectoderm, were isolated and cultured they developed into ciliated embryoids that were morphologically polarized. In animal explants from S. purpuratus, the aboral ectoderm-specific Spec1 gene was activated at the same time as in control embryos and at relatively high levels. The Spec1 protein was restricted to the squamous epithelial cells in the embryoids suggesting that an oral-aboral axis formed and aboral ectoderm differentiation occurred correctly. However, the Ecto V protein, a marker for oral ectoderm differentiation, was detected throughout the embryoid and no stomodeum or ciliary band formed. These results indicated that animal explants from S. purpuratus were autonomous in their ability to form an oral-aboral axis and to differentiate aboral ectoderm, but other aspects of ectoderm differentiation require interaction with vegetal blastomeres. In contrast to S. purpuratus, aboral ectoderm-specific genes were not expressed in animal explants from L. pictus even though the resulting embryoids were morphologically very similar to those of S. purpuratus. Recombination of the explants with vegetal blastomeres or exposure to the vegetalizing agent LiCl restored activity of aboral ectoderm-specific genes, suggesting the requirement of a vegetal induction for differentiation of aboral ectoderm cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Inositol-1,4,5-trisphosphate injection mimics fertilization potentials in sea urchin eggs

1986 ◽  
Vol 250 (2) ◽  
pp. C340-C344 ◽  
Author(s):  
B. E. Slack ◽  
J. E. Bell ◽  
D. J. Benos
Keyword(s):  
Membrane Potential ◽  
Partial Response ◽  
Sea Urchin ◽  
Time Course ◽  
Strongylocentrotus Purpuratus ◽  
Sea Urchin Eggs ◽  
Low Calcium ◽  
Inositol 1,4,5 Trisphosphate

The effects of inositol-1,4,5-trisphosphate (IP3) and of diacylglycerol (DAG) and its analogues on the membrane potential of eggs from the sea urchin Strongylocentrotus purpuratus were examined. Injection of IP3 into eggs resulted in a change in membrane potential that was similar in magnitude and time course to the fertilization potential elicited by sperm attachment. In low-calcium seawater, IP3 injection elicited a partial response. DAG and its analogues phorbol myristyl acetate and 1-oleoyl-2-acetylglycerol did not affect membrane potential either when applied by perfusion or when injected. The results indicate that IP3, but not DAG or its analogues, may be involved in the generation of the fertilization potential triggered by the interaction of sperm with sea urchin eggs.

Displacement of Valine From Intact Sea-Urchin Eggs by Exogenous Amino Acids

1968 ◽  
Vol 3 (4) ◽  
pp. 515-527
Author(s):  
J. PIATIGORSKY ◽  
A. TYLER
Keyword(s):  
Amino Acids ◽  
Glutamic Acid ◽  
Sea Urchin ◽  
Sea Water ◽  
The Other ◽  
Metabolic Product ◽  
Primary Factor ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
Neutral Amino Acids

Unfertilized and fertilized eggs of the sea urchin Lytechinus pictus were preloaded with [14C]valine and exposed to individual solutions of each of the twenty ‘coded’ [12C]amino acids in artificial sea water. After 1 h incubation the amount of radioactivity in the medium was determined. The radioactivity was effectively displaced by most of the other neutral [12C]amino acids that are known to compete with valine for uptake. A chromatographic test with fertilized eggs showed the displaced radioactivity to be [14C]valine and not some metabolic product. Addition of acidic, basic or some neutral amino acids that are known to be poor inhibitors of valine uptake did not cause significant quantities of label to appear in the medium. For the unfertilized eggs, the concentration of acid-soluble label remained many hundreds of times greater in the egg fluid than in the sea water. Tests indicated that efflux of [14C]valine and subsequent competition for re-entry is a primary factor responsible for the displacement phenomenon. That this may not be the sole factor is suggested by the fact that some amino acids that are known to be powerful inhibitors of valine uptake were found to be only weak displacers of [14C]valine. Neither [14C]arginine nor [14C]glutamic acid were displaced in significant amounts from preloaded unfertilized or fertilized eggs by any of the tested [12C]amino acids. Attempts were made to utilize the displacement of [12C]valine to elevate the incorporation of [14C]valine and of other labelled amino acids into protein by intact eggs. Unfertilized and fertilized eggs were pretreated with related [12C]amino acids and then exposed to [14C]valine or a mixture of [14C]amino acids. The results varied in the different tests, ranging from no significant increase to 2-fold.

Cortical Granule Exocytosis and Fertilization Coat Elevation is Reduced in Aging Sea Urchin Eggs

2000 ◽  
Vol 6 (S2) ◽  
pp. 966-967
Author(s):  
Amitabha Chakrabarti ◽  
Heide Schatten
Keyword(s):  
Plasma Membrane ◽  
Sea Urchin ◽  
Sea Water ◽  
Secretory Granules ◽  
Cortical Granule ◽  
Cortical Granules ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
Cortical Granule Exocytosis ◽  
Granule Secretion

Cortical granules are specialized Golgi-derived membrane-bound secretory granules that are located beneath the plasma membrane in unfertilized sea urchin eggs. Upon fertilization cortical granules discharge in a reaction induced by calcium and release their contents between the plasma membrane and a thin vitelline layer that lines the plasma membrane. Microvilli at the plasma membrane elongate incorporting cortical granule membranes during elongation. The vitelline layer elevates and becomes the egg's fertilization coat that hardens and serves as physical block to polyspermy. While we do not understand the precise mechanisms that participate in cortical granule discharge it is believed that actin plays a role in this process. Because actin and calcium metabolism is affected in aging cells we investigated if cortical granule secretion is affected in aging sea urchin eggs.Lytechinus pictus eggs were obtained by intracoelomic injection of 0.5M KCI to release the eggs into sea water at 23°C.

scholarly journals Activation of sea urchin eggs by inositol phosphates is independent of external calcium

1988 ◽  
Vol 252 (1) ◽  
pp. 257-262 ◽  
Author(s):  
I Crossley ◽  
K Swann ◽  
E Chambers ◽  
M Whitaker
Keyword(s):  
Sea Urchin ◽  
Calcium Ions ◽  
Sea Water ◽  
Inositol Phosphates ◽  
Inositol Phosphate ◽  
Egg Activation ◽  
Lytechinus Variegatus ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
External Calcium

We investigated the contribution of external calcium ions to inositol phosphate-induced exocytosis in sea urchin eggs. We show that: (a) inositol phosphates activate eggs of the sea urchin species Lytechinus pictus and Lytechinus variegatus independently of external calcium ions; (b) the magnitude and duration of the inositol phosphate induced calcium changes are independent of external calcium; (c) in calcium-free seawater, increasing the volume of inositol trisphosphate solution injected decreased the extent of egg activation; (d) eggs in calcium-free sea water are more easily damaged by microinjection; microinjection of larger volumes increased leakage from eggs pre-loaded with fluorescent dye. We conclude that inositol phosphates do not require external calcium ions to activate sea urchin eggs. This is entirely consistent with their role as internal messengers at fertilization. The increased damage caused to eggs in calcium-free seawater injected with large volumes may allow the EGTA present in the seawater to enter the egg and chelate any calcium released by the inositol phosphates. This may explain the discrepancy between this and earlier reports.

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