In vitro release of biological active molecules during the clotting reaction in Holothuria polii

Abstract

1. 1. In the holothuroid Holothuria polii clotting of coelomocytes occurs in vitro 2. 2. The results show that in Edds' isotonic salt solution clot reaction follows three phases: an immediate agglutination of amebocytes and spherule cells (first phase), a subsequent larger aggregation of the clump (second phase) and, finally, a contraction of the whole aggregated mass in a clot (third phase). 3. 3. The presence of 30 mM EDTA in Edds' isotonic salt solution abrogates the clotting reaction indicating that calcium or magnesium dependent clotting occurs. 4. 4. The presence of reducing agents (EIS-caffeine and 2-mercaptoethanol) did not prevent initial aggregation of coelomocytes but it inhibited the following steps in the reaction. 5. 5. Agglutinating molecules are not released in EIS-EDTA suggesting that clumping could be mediated by these factors. 6. 6. Defensive molecules, hemolysin and lysozyme, are also produced during the clotting reaction. 7. 6. At least for lysozyme, the anti-clotting agents used do not inhibit its release.