Novel polyamines in insects and spiders

• 1.1. Diaminopropane, putrescine, norspermidine, spesrmidine, norspermine and spermine were commonly found in the larval silk gland and head of Bombyx mori, Antheraea yamamai and Galleria mellonella.
• 2.2. The cockroach Periplaneta americana contained thermospermine, caldopentamine, caldohexamine, homospermidine, aminopropylhomospermidine and aminobutylhomospermidine (homospermine) in addition to the common polyamines. This is the first report to show the occurrence of the homospermidine derivatives and a hexamine in insects.
• 3.3. In addition to thermospermine, caldopentamine, agmatine, histamine, cadaverine and other usual polyamines, three cadaverine-derivatives, aminopropylcadaverine, bis(aminopropyl)cadaverine and aminopentylnorspermidine, were detected in the silk gland and head of the spiders, Nephila clavata and Araneus ventricosus. The occurrence of aminopropyl derivatives of of cadaverine has never been reported in animals.

     

Chemoreception in the shrimp,Palaemonetes pugio: The role of amino acids and betaine in elicitation of a feeding response by extracts

• 1.1. Substances of less than ca. 1000 molecular weight present in extracts of blue crab, oyster, sea urchin and mullet were the principal stimulants of a feeding response in the shrimp, Palaemonetes pugio.
• 2.2. Synthetic mixtures of amino acids and betaine, formulated on the basis of the composition of the extracts, were found to account for 60–100% of the activity of extracts of crab, oyster and sea urchin but for only ca. 30% of the activity of mullet extract.
• 3.3. When tested alone, betaine had only a modest stimulatory capacity; yet this substance made a very significant contribution to the activity of the mixtures.
• 4.4. Assays of betaine alone, or of certain mixtures of amino acids, yielded unusual “bell-shaped” dose-response relationships.

     

Fluxes of dissolved amino acids between sea water and Echinaster

• 1.1. Measurement of free amino acid (primary amine) influx and efflux into the starfish, Echinaster, were accomplished utilizing improved methods of sea water purification and analysis.
• 2.2. Specimens placed in amino acid depleted sea water (5 × 10⁻⁸ M) demonstrated net release as measured with the fluorescamine method. Similarly, specimens placed in the same water to which amino acid mixtures had been reintroduced to normal levels demonstrated net uptake.
• 3.3. A mathematical model indicated an equilibrium amino acid concentration (when influx equals efflux) of 5.26 × 10⁻⁷ M, or about one fourth the level of natural sea water.
• 4.4. Since at normal environmental levels (20.65 × 10⁻⁷ M) net flux is inward by a ratio of nearly 4-1, it is concluded that the previous suggestions of some workers that such would not be the case for marine invertebrates are no longer valid.
• 5.5. The net uptake of amino acid from environmental levels would account for 5.67% of the measured total respiration if all were being metabolized.
• 6.6. This figure appears to be in line with the previously developed hypothesis that the epidermis largely obtains its nutrition directly from the environment. However, the real benefit of the uptake mechanism may be to prevent loss of the body amino acid pools.

     

Heart rates of sea snakes diving in the sea

• 1.1. Classical diving bradycardia was not evident in sea snakes diving in the sea.
• 2.2. Activity increases heart rate both in apneic and in breathing snakes; there is a breathing tachycardia in the laboratory and in the sea.
• 3.3. Heart rates obtained from animals diving in the sea (26 m depth) were similar to those obtained under laboratory conditions if allowances are made for activity differences.
• 4.4. Handling and stress increases heart rates of sea snakes.

     

Characterization of two major non-histone protein fractions from chromatin of sea urchin embryos

• 1.1. Chromatin of sea urchin embryos was chromatographed on a hydroxyapatite column, and the two largest fractions of proteins desorbed with 0.05 M and 0.2 M Na-phosphate, pH 6.8, were comparatively characterized.
• 2.2. The mol. wt ranges of these fractions were about 18,000 and 14,000 daltons, respectively, at both molarities of the phosphate buffer.
• 3.3. Both fractions at each elution molarity were found to be acidic, and to differ somewhat in their respective amino acid compositions. No important differences related to the stage of development could be detected in this respect.
• 4.4. Upon re-electrophoresis both large fractions resolved in one major and three minor bands. The amino acid composition of the major band was different for the blastula and gastrula chromatin.
• 5.5. It is likely that these fractions contain glycoprotein subunits.

     

Short-term osmotic responses of cells and tissues of the sea anemone,Condylactis gigantea

• 1.1. Cells of tentacles and body wall of the sea anemone Condylactis gigantea behaved as simple osmometers during 5hr exposure to 50, 67, 83, 100 and 125% sea-water.
• 2.2. All intracellular water appeared to be osmotically active.
• 3.3. Cell sodium, chloride and total osmolyte content remained invariable, with taurine decreasing and potassium increasing as sea-water concentration was reduced.
• 4.4. Tissues, as a whole, exhibited a pseudoregulatory response to changes in salinity as the large and osmotically inert extracellular space buffered volume changes to a considerable extent.

     

A halogenated amino acid-containing sperm activating peptide and its related peptides isolated from the egg jelly of sea urchins,Tripneustes gratilla,Pseudoboletia maculata,Strongylocentrotus nudus,Echinometra mathaei and Heterocentrotus mammillatus

• 1.1. Twenty-eight peptides were isolated from the egg jelly of sea urchins, Tripneustes gratilla, Pseudoboletia maculata, Strongylocentrotus nudus, Echinometra mathaei (type A and B) and Heterocentrotus mammillatus and their amino acid sequences were determined.
• 2.2. Two of the peptides obtained from T. gratilla egg jelly possessed a bromophenylalanine (Br-Phe) residue in their sequences (Gly-(Br-Phe)-Asn-Leu-Asn-Gly-Gly-Gly-Val-Gly and Gly-(Br-Phe)-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly).
• 3.3. All of the peptides elevated cyclic GMP concentrations in the spermatozoa of the respective sea urchin and caused a shift in the apparent mol. wt of a major sperm protein of the respective sea urchin.
• 4.4. They stimulated respiration rates of the spermatozoa of Hemicentrotus pulcherrimus as well as their own species.
• 5.5. One-half maximal concentrations of the peptides for respiratory stimulation of H. pulcherrimus spermatozoa were between 10⁻¹¹ M and 10⁻⁹ M except a methionine-containing peptide which was about 10⁻⁷ M.

     

Time related changes in the free amino acid pool of the sea anemone,Bunodosoma cavernata,during salinity stress

• 1.1. The sea anemone, Bunodosoma cavernata, is a relatively eurybaline cnidarian tolerating salinities from 12 to 40%.
• 2.2. Taurine, glutamic acid and aspartic acid all showed some increases with increased salinity.
• 3.3. The amino acid showing the greatest accumulation under high salinity conditions was β-alanine which increased 28-fold from 1.5 to 41.9 μmol/g dry weight when salinity was raised from 26 to 40%.
• 4.4. When B. cavernata was subjected to increased salinity, β-alanine was rapidly accumulated and reached maximum levels within 4 days.
• 5.5. When salinity was dropped from 36 to 26%0, β-alanine concentrations dropped from 15 to 2 μmol/g dry weight in 2 days.

     

Collagen diversity in the sea urchin,strongylocentrotus purpuratus

• 1.1. Pepsin insensitive fragments of collalgen extracted from tube feet and peristome have α 1 and α 2 bands that differ in apparent molecular weights from each other and from human type 1 collagen.
• 2.2. A monoclonal antibody that reacts with the ξ I band and a low molecular weight fragment of tube foot collagen does not react with either peristome collagen or human type I collagen.
• 3.3. Measurements of the axial periodicity of native fibers of tube foot and peristone collagens indicate they have D values that differ significantly from each other and from reported values of vertebrate type I collagen.
• 4.4. We propose that there are diverse and specialized types of collagen in sea urchins that are heterogeneously distributed in the extracellular matrix of different tissue.

     

N-terminal substitution of some sea cucumber hemoglobins

• 1.1. Each of the four major globins found in the erythrocytic hemoglobins of the sea cucumber Molpadia arenicola have been shown to be N-acetylated.
• 2.2. The hemoglobins from the sea cucumbers M. oölitica, Cucumaria rubra, Thyonella pervicax and Pentacta pygnaea were also shown to be blocked at the N-terminus.

     

Occurrence of d-alanine in the eggs and the developing embryo of the sea urchin Paracentrotus lividus

• 1.1. d-Alanine has been found in appreciable amounts in the eggs and embryos of the sea urchin Paracentrotus lividus.
• 2.2. The content of d-alanine, expressed as pmol/egg or embryo, is 1.32 in the egg, 0.81 in the blastula, 0.54 in the gastrula and 0.60 in the pluteus.
• 3.3. The percentage of d-alanine with respect to the total alanine (d + l) decreases during embryonic development.
• 4.4. d-Amino acid oxidase, d-alanine transaminase and d-alanine racemase activities were found neither in eggs nor in embryos.
• 5.5. Therefore, it does not appear likely that d-alanine is subject to oxidative metabolism.
• 6.6. The decrease in this d-amino acid during development may be due to its utilization in the synthesis of a more complex molecule.

     

Pigmentation in the sea anemone Corynactis californica

• 1.1. There is great variability in the color of the sea anemone, Corynactis californica (Calgren, 1936). Studies of the columns of seven different color varieties showed no major differences in type or concentration of carotenoids. An astaxanthin ester and an unidentified carotene were isolated.
• 2.2. The isolated carotenoids do not appear to be responsible for the variety of colors in Corynactis.
• 3.3. The color variation of C. californica is more probably dependent on an unidentified, water-soluble pigment similar to a bile-like pigment, calliactine, isolated from the sea anemone, Sagartia parasitica (Couch, 1838).

     

Two acid phosphatases in sea urchin eggs and embryos

• 1.1. Two types of acid phosphatases from sea urchin eggs and embryos were studied in three Japanese species, Hemicentrotus pulcherrimus, Anthocidaris crassispina and Pseudocentrotus depressus.
• 2.2. Acid phosphatase type 1, designated AcP-1, hydrolysed only flavin mononucleotide besides p-nitrophenylphosphate. The activity of AcP-1 was not inhibited by NaF and tartrate. This enzyme showed molecular weight between 14,000 and 16,000 by gel filtration through Sephadex G-75.
• 3.3. The higher molecular weight type of acid phosphatase, designated AcP-2, showed relatively high substrate specificity toward ADP and ATP. Molecular weight of AcP-2 ranged from 42,000 to 48,000 by gel filtration through Sephadex G-100.
• 4.4. Some properties of AcP-2 from Sphaerechinus granularis embryos are also described.

     

Preparation and properties of mitochondria from tissues of the sea mussel Mytilus edulis L

• 1.1. A method is described for the preparation of coupled mitochondria from hepatopancreas, mantle and adductor muscle of the sea mussle, Mytilus edulis.
• 2.2. NADH is a non-penetrant, whereas succinate, glutamate and malate plus pyruvate have a clear stimulatory effect on respiration. Proline does not stimulate respiration of sea mussel mitochondria.
• 3.3. No P/0 ratios could be calculated as after the addition of ADP the mitochondria remain in the active state (state III), even after enough oxygen is consumed for complete phosphorylation. The reason for this observation is discussed.
• 4.4. Mitochondrial densities based on cytochrome b levels of two tissues of the mussel are compared to rat liver. Within the mussel the mantle contains twice the amount of cytochrome of adductor muscle.

     

Fate of ingested leucine and glucose in the sea star,Asterias rubens,during its annual reproductive cycle

• 1.1. The distribution of radiolabel from L-leucine [¹⁴C-UL] and D-glucose [¹⁴C-UL] was measured in the sea star Asterias rubens at 1, 6 and 24 hr after oral administration.
• 2.2. Incorporation of the label from both compounds was observed in pyloric caeca, coelomocytes and ovaries even after an incubation time of 1 hr.
• 3.3. Highest incorporation from both precursors was found in proteins, while substantial radioactivity was present in the amino acids, organic acids and neutral components. Lipids were hardly labelled from leucine and only slightly from glucose.
• 4.4. Radioactivity in proteins and lipids increased with increasing incubation time. No significant differences were found in the distribution patterns of radiolabel during the reproductive cycle.
• 5.5. The data obtained are discussed in terms of current knowledge on the translocation of nutrients in echinoderms.

     

Water-soluble Ca2+-calmodulin-binding proteins in embryo of the sea urchin Strongylocentrotus intermedius

• 1.1. About 0.3–0.4% of total water-soluble protein extracted from sea urchin embryos at the two-cell and early-gastrula stages was Ca²⁺-dependently bound to immobilized calmodulin.
• 2.2. SDS-PAGE of calmodulin-binding proteins revealed at least 20 polypeptides ranging from 200 to 15.5 kDa, and 70–80% of the protein belonged to a dozen major polypeptides. Polypeptides of 70, 55, 50, 45 and 18 kDa seemed to be the same as those that were detected earlier (Iwasa and Mohri, J. Biochem.94, 575–587, 1983).
• 3.3. The polypeptide spectrum of calmodulin target proteins changed significantly, e.g. the major polypeptides of 70 and 41 kDa increased, and the 200 and 43 kDa polypeptides decreased sharply during development from the two-cell to the early-gastrula stage.
• 4.4. According to our estimates, the molar concentrations of the calmodulin and targets were close enough, and therefore the Ca²⁺ signal should depend on the spatial-temporal distribution of free calmodulin in the cells.

     

Microheterogeneity of late histones in larval stages of sea urchin development

• 1.1. The total histone complement of early plutei were compared with that of intermediate and late larvae of the sea urchin Tetrapygus niger.
• 2.2. Electrophoretic comparison indicates that there are quantitative and qualitative shifts of the five classes throughout late larval development.
• 3.3. The strong similarity in the amino acid composition of total histones isolated from early, intermediate and late plutei indicates that the observed electrophoretic heterogeneity is due to post-translational modifications.

     

Comparative study of proteolytic enzymes in the digestive tracts of the european sea bass and hybrid striped bass reared in freshwater

• 1.1. A comparative study of the proteolytic activity in four different sections of the digestive tracts of the European sea bass (Dicentrarchus labrax) and hybrid striped bass (Morone chrysops × M. saxatilis) reared in freshwater revealed minor differences between these fish.
• 2.2. Tryptic activity plays a major role in the proteolytic process in both fish.
• 3.3. The activity of seven intestinal proteolytic enzymes was detected utilizing a combination of specific substrates and inhibitors.
• 4.4. High levels of proteolytic activity were detected in both the proximal and distal sections of the fish intestine at a high pH range (9–10).
• 5.5. In situ monitoring of pH levels revealed a lower pH level in the intestinal proximal section of hybrid striped bass compared with the distal section.
• 6.6. In contrast, higher pH levels were detected at the proximal compared with the distal sections of D. labrax intestine.

     

Lipid composition of the red and green forms of Actina equinia from the black sea

• 1.1. The fatty-acid composition of the red and green forms of Actinia equina from the Black Sea have been determined by methods involving silver-ion HPLC and GC-MS.
• 2.2. The fatty acid compositions of both forms of A. equina resemble those of most marine invertebrates. Substantial amounts of C₂₀ and C₂₂ polyunsaturated fatty acids were found.
• 3.3. Balck Sea Actenia equina contains a large amount of plasmalogens, mainly phospotidylcholine and phosphatidylserine plasmalogens.
• 4.4. The red form of A. equina contains more arachidonic acid in glycolipid fraction and more phosphatidylethanolamine and its plasmalogen, while the green from contains more sphingomyelin. These differences are an indication that the species A. equina can be divided into two subspecies—green and red.

     

Purification and characterization of arylsulfatase from sea urchin (Hemicentrotus pulcherrimus) embryos

• 1.1. Arylsulfatase was extracted from sea urchin (Hemicentrotus pulcherrimus) plutei and purified to electrophoretical homogeneity by means of DEAE-cellulose, acetone fractionation and Sepharose CL-6B, successively.
• 2.2. The molecular weight of this enzyme was approx, 670,000. The molecular weight of a single subunit was approx. 63,000. The K_m value for p-nitrophenyl sulfate was 0.59 mM.
• 3.3. This enzyme was competitively inhibited by the sulfate ion and was classified as the type II arylsulfatase. The pH optimum was between 5.0 and 6.0.