Michèle Durliat

ACTION OF VARIOUS ANTICOAGULANTS ON HEMOLYMPHS OF LOBSTERS AND SPINY LOBSTERS

Effects of anticoagulants on hemolymph coagulation of lobsters are reported. Effects of some inhibitors on plasma clotting change in relation to molting stages of animals. Variations of electrophoretic and immunochemical protein patterns are analyzed. Complete coagulation inhibition shows that the plasma coagulogen exists as a mixture of soluble uncovalently crosslinked polymers. In lobsters the molecular weight of the lightest visible circulating unit is approximately 420,000 daltons. The weight interval between each of the four lighter polymers is about 200,000 daltons. Serine protease inhibitors impair plasma clotting and interact with cellular clumping.

COAGULATION IN THE CRAYFISH, ASTACUS LEPTODACTYLUS: ATTEMPTS TO IDENTIFY A FIBRINOGEN-LIKE FACTOR IN THE HEMOLYMPH

1. A series of tests were conducted to determine whether or not the hemolymph of the crayfish Astacus leptodactylus contains a plasma coagulation factor. 2. The total protein amount is higher in the plasma than in the serum. 3. Serum and plasma do not exhibit similar electrophoretic banding patterns. Plasma contains one band or a series of supplementary fractions with a high molecular weight. 4. Electrophoregrams of plasmatic clottable extract, obtained by the classical methods employed in crustacean serology, show a main fraction or a series of polymers with the same electrophoretic behavior as the additional fractions seen on the plasma pattern. 5. This solution clots when treated with CaCl2 and a cellular extract. 6. Immunoelectrophoresis demonstrates the presence of a clottable protein precipitate line in plasma, but this protein also gives a very faint similar line in serum.

Changes in water-soluble proteins from integument of Astacus leptodactylus during the molt cycle

Abstract 1. 1. Changes in soluble proteins from freshwater crayfish integument have been studied by electrophoresis and two-dimensional immunoelectrophoresis from anecdysis stage C4 to early metecdysis stage A. 2. 2. Two integumental fractions present in the hemolymph during the same period show important variations. 3. 3. Fraction 1 which runs in agarose as the bromphenol blue does is important from later C4 to D1′, decreases mainly after the stage D2 and then reappears after the molt. It seems to be a very acidic lipoprotein of small molecular weight, present only in small amounts in the blood. 4. 4. Present in the hemolymph at a considerable amount during the premolt, fraction 2 exists in the integument during all the premolt long but rises mainly in D1, D2 and A. 5. 5. In the integument, fibrinogen-like factor appears at a rate relatively more important than in the plasma. 6. 6. On the other hand, at least one or two integument proteins can be found. One of them is one crustacyenin.

COMPARISON OF THE GRADIENT OF SETAL DEVELOPMENT OF UROPODS AND OF SCAPHOGNATHITES IN ASTACUS LEPTODACTYLUS

1. Characterization of the different stages of the molt cycle of Astacus leptodactylus has been established using morphological criteria of the setae observed through the uropod. This method is paraticularly useful for the scientist who wants to get replicable, physiological data without disturbing the animals.2. Stages determined from uropod studies agree with those obtained from comparative studies of scaphognathites.3. According to the seasons, a more or less quiet intermolt phase is observed. The animals either directly enter the premolt phase or are blocked in stage C4 or early DO. Therefore, the morphological stage DO seems to be heterogenous, and apolysis is not necessary to the onset of premolt.4. A single mode of setal formation seems evidenced. Moreover, setae with compound or connected matrices are seen in the scaphognathite.5. Number and mobility of blood cells in the uropods are related to the investigated stage. During the premolt stages hemocytes, especially granulocytes, are observed withi...

Occurrence of plasma proteins in ovary and egg extracts from Astacus leptodactylus

Abstract 1. 1. In both ovary and egg extracts from crayfish, several components show immunological identities with the blood proteins. 2. 2. Haemocyanin is always detected but appears more heterogeneous in the ovaries than in other compartments. 3. 3. Fraction termed Fb, present in the haemolymph and hepatopancreas, is found to an appreciable amount. 4. 4. Both coagulogen and fraction Fa are not observed. 5. 5. Two main lipoprotein fractions exist in the haemolymph of vitellogenic females, in the ovaries and in the eggs. They appear identical throughout the vitellogenic cycle in all samples, but each of them is immunochemically different from the other. 6. 6. The synthesis of embryonic components is discussed.

Proteins of aqueous extracts from the hepatopancreas of Astacus leptodactylus—1. Changes in proteins during the molt cycle

Abstract 1. 1. Total protein content in the crayfish hepatopancreas exhibits a certain rhythm: the level rhythm: the level falls in postmolt, rises in stage C4, decreases again from D0 to D1″ and then increases in D2–D3. 2. 2. Electrophoregrams of soluble extracts show some proteins with a high mol. wt (about 400,000 wt (about 400,000 daltons) but many fractions have a small mol. wt (below 100,000 daltons). 3. 3. Four proteases and three amylases have been observed. 4. 4. Four groups of fractions exhibit evident asynchronous quantitative variations throughout the molting cycle. 5. 5. Several fractions cross-react with an antihemolymph serum.

Cuticular proteins in the crayfish Astacus leptodactylus—I. Main components

Abstract 1. 1. A few specific proteins were extracted from the hardened calcified cuticle of Astacus leptodactylus. In their native state, all had a high mol. wt and a pI between 4 and 6. 2. 2. The native crustacyanins were blue and appeared as closely related α and γ forms. These two crustacyanins easily dissociated into different transitory subunits and then into a single β crustacyanin. 3. 3. The Cut 2 fraction also dissociated but the Cut 1, Cut 3 and Cut 3′ fractions were very stable. 4. At least four proteins originating from the blood were found in small amounts in the calcified shell, including hemocyanin, plasma coagulogen, molt-related F2 fraction and an F4 fraction. The role of these components is discussed.

Cuticular protein analysis from Astacus leptodactylus—1. Electrophoretical studies of the soluble cuticular proteins

Abstract 1. 1. A sequence of different solvents was used to extract the non-covalently bound proteins from the calcified cuticle of crayfish. 2. 2. Electrophoretical pattern of the major bands from various extracts differed between themselves and from those of plasma or soft integument extracts. 3. 3. Up to 34 proteins were visualized. Most of them, including the major ones, showed a molecular weight beneath 50,000 and an isoelectric point between pH 3.5–5.0. 4. 4. Calcium stabilized many proteins, especially crustacyanin. Use of detergent showed that sclerotin could rise from arthropodins.

Cuticular proteins in the crayfish Astacus leptodactylus—II. Relationships between proteins in plasma soft integument and calcified cuticle

Abstract 1. 1. Both the number and amount of plasma proteins were larger in extracts obtained from friable cuticles than in those prepared from hardened cuticles. In contrast, the changes occurring in cuticle throughout the molting cycle affected its total protein content but not the number of proteins present. 2. 2. Crustacyanins appear to be immunochemically identical in both the soft integument and hardened shell. The dissociated β form of crustacyanin displayed partial antigenic identities with the native α and γ forms. The Cut 2 fraction changed during transit through different compartments. 3. 3. The other major cuticular proteins Cut 1, Cut 3 and Cut 3′ were found in calcified carapace extracts only. 4. 4. The true significance of F2 molt-related fraction is discussed.

Proteins of aqueous extracts from the hepatopancreas of Astacus leptodactylus—2. Immunological identities of proteins from hepatopancreas and blood

Abstract 1. 1. About 15 proteins of aqueous extracts from the crayfish hepatopancreas exhibit cross reactions with an antihemolymph serum. 2. 2. Five main evident fractions corresponding to the blood proteins are observed. Both hemocyanin and coagulogen are present with a little relative amount. Components termed as Fa and Fb are detected with a concentration twice smaller than in the hemolymph. 3. 3. Both molt and vitellogenesis-related proteins are not found. 4. 4. Other proteins show only some partial antigenic identities with those of the hemolymph. 5. 5. Role of hepatopancreas in the synthesis of some blood proteins is discussed.