Induction of Fertilization Membrane Formation and Cyanide-insensitive Respiration in Sea Urchin Eggs by the Treatment with Dimethylsulfoxide Followed by an Incubation in an Ice Bath. (dimethylsulfoxide/sea urchin egg/fertilization membrane/respiration/verapamil)

From electron microscope studies of thin sections the sea urchin egg is known to be surrounded by the peripheral membrane system which is made up of the outer coat (vitelline membrane), which elevates from an egg surface after fertilization and becomes a part of the fertilization membrane, and the plasma membrane. In these experiments an effort has been made to isolate plasma membranes of sea urchin eggs and these isolated membranes were observed in the electron microscope.The vitelline membrane of the eggs from the sea urchin Strongylocentrotus purpuratus was at first digested away by the treatment with 0.02% trypsin in 0.01 M Tris-HCl buffer (pH 8.0) for 5 minutes at 28°C. The plasma membranes were then isolated according to the method of Song et al. which was used for the isolation of rat liver plasma membranes. The vitelline membrane-free eggs were gently homogenized in 10-3 M NaHC03 (pH 7.5) and freed membranes were collected by centrifugation over a discontinuous sucrose gradient preparation.

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Metabolic similarities between fertilization and phagocytosis. Conservation of a peroxidatic mechanism.

Journal of Experimental Medicine ◽

10.1084/jem.149.4.938 ◽

1979 ◽

Vol 149 (4) ◽

pp. 938-953 ◽

Cited By ~ 48

Author(s):

S J Klebanoff ◽

C A Foerder ◽

E M Eddy ◽

B M Shapiro

Keyword(s):

Sea Urchin ◽

Polymorphonuclear Leukocytes ◽

Cortical Granules ◽

Sea Urchin Eggs ◽

Sea Urchin Egg ◽

Hatching Process ◽

Fertilization Membrane ◽

Catalyzed Reactions ◽

Kinetics Of ◽

Estrogen Binding

At the time of fertilization, sea urchin eggs release a peroxidase which, together with H2O2 generated by a respiratory burst, is responsible for hardening of the fertilization membrane. We demonstrate here that the ovoperoxidase of unfertilized eggs is located in cortical granules and, after fertilization, is concentrated in the fertilization membrane. Fertilization of sea urchin eggs or their parthenogenetic activation with the ionophor A23187 also results in (a) the conversion of iodide to a trichloroacetic acid-precipitable form (iodination), (b) the deiodination of eggs exogenously labeled with myeloperoxidase and H2O2, (c) the degradation of thyroxine as measured by the recovery of the released radioiodine at the origin and in the inorganic iodide spot on paper chromatography, and (d) the conversion of estradiol to an alcohol-precipitable form (estrogen binding). The iodination reaction and the binding of estradio occurs predominantly in the fertilization membrane where the ovoperoxidase is concentrated. From the estimation of the kinetics of incorporation of iodine, we determine that the peroxidative system is active for 30 min after fertilization, long after hardening of the fertilization membrane is complete. Most of the bound iodine is lost during the hatching process. Iodination of albumin is catalyzed by the material released from the egg during fertilization, when combined with H2O2 and iodide. Iodination, thyroxine degradation, and estradiol binding are inhibited by azide, cyanide, aminotriazole, methimazole, ascorbic acid and ergothioneine, all of which can inhibit peroxidase-catalyzed reactions. These responses of the sea urchin egg to fertilization are strikingly similar to the changes induced in polymorphonuclear leukocytes by phagocytosis and, in both instances, a peroxidative mechanism may be involved.

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Nicotinic acid-adenine dinucleotide phosphate mobilizes Ca2+ from a thapsigargin-insensitive pool

Biochemical Journal ◽

10.1042/bj3150721 ◽

1996 ◽

Vol 315 (3) ◽

pp. 721-725 ◽

Cited By ~ 125

Author(s):

Armando A. GENAZZANI ◽

Antony GALIONE

Keyword(s):

Nicotinic Acid ◽

Sea Urchin ◽

Cross Talk ◽

Inositol Trisphosphate ◽

Sea Urchin Eggs ◽

Bivalent Cations ◽

Sea Urchin Egg ◽

Different Characteristics ◽

Degree Of Overlap

Nicotinic acid–adenine dinucleotide phosphate (NAADP) is a novel intracellular Ca2+ releasing agent recently described in sea-urchin eggs and egg homogenates. Ca2+ release by NAADP is independent of that induced by either inositol trisphosphate (InsP3) or cyclic adenosine dinucleotide phosphate (cADPR). We now report that in sea urchin egg homogenates, NAADP releases Ca2+ from a Ca2+ pool that is distinct from those that are sensitive to InsP3 and cADPR. This organelle has distinct Ca2+ uptake characteristics: it is insensitive to thapsigargin and cyclopiazoic acid, but maintenance of the pool shows some requirement for ATP. Although the different Ca2+ pools have different characteristics, there appears to be some degree of overlap or cross-talk between the NAADP- and cADPR/InsP3-sensitive Ca2+ pools. Ca2+-induced Ca2+ release is unlikely to account for the apparent overlap between stores, since NAADP-induced Ca2+ release, in contrast with that stimulated by cADPR, is not potentiated by bivalent cations.

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Ryanodine Activates Sea Urchin Eggs. (sea urchin egg/activation/ryanodine/inositol phosphate/heparin)

Development Growth & Differentiation ◽

10.1111/j.1440-169x.1992.00037.x ◽

1992 ◽

Vol 34 (1) ◽

pp. 37-42 ◽

Cited By ~ 18

Author(s):

C. Sardet ◽

I. Gillot ◽

A. Ruscher ◽

P. Payan ◽

J.-P. Girard ◽

...

Keyword(s):

Sea Urchin ◽

Inositol Phosphate ◽

Egg Activation ◽

Sea Urchin Eggs ◽

Sea Urchin Egg

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Some Experiments on Sea-urchin Eggs with Desoxynucleic Acids from Sea-urchin Sperms

Development ◽

10.1242/dev.1.3.261 ◽

1953 ◽

Vol 1 (3) ◽

pp. 261-262

Author(s):

Sven Hörstadius

Keyword(s):

New York ◽

Sea Urchin ◽

Sea Water ◽

Water Injection ◽

Dark Field ◽

The Other ◽

Columbia University ◽

Sea Urchin Eggs ◽

King's College ◽

Fertilization Membrane

Dr. I. Joan Lorch, of King's College, London, and I have made some experiments on sea-urchin eggs with desoxynucleic acids (DNA) prepared from sperms of several sea-urchin species by Professor Erwin Chargaff, of Columbia University, New York. Unfertilized eggs did not react when put into a solution of DNA in sea-water. Injection of a small amount of DNA dissolved in Callan's solution had the following consequences. If the DNA did not mix with the cytoplasm but remained as a distinct droplet, the egg could be fertilized. The droplet moved slowly towards the surface and ran out of the egg. This sometimes only occurred after several cleavages. Such eggs developed normally. If, on the other hand, the DNA mixed with the cytoplasm the egg became activated. A fertilization membrane was raised. The surface layer in dark field changed in colour from yellow to white as is the case upon fertilization.

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The Respiratory and Glycolytic Enzymes of Sea-Urchin Eggs

Journal of Experimental Biology ◽

10.1242/jeb.31.2.208 ◽

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.

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