ACROSOME REACTION IN SPERM FROM MUD CRAB (SCYLLA SERRATA)

摘要: The nonmotile sperm with a large and complex acrosome body is well known characteristic of decapod crustaceans. reaction this kind more than others. Although lot work focused on the structure crustaceans has been published, little information available about how to artificially induce high rate success. ultrastructure mature spermatogenesis Scylla serrata , commercially important species crab, have reported in our previous work. As part ongoing research, we investigated optimal conditions for artificial induction observed process present study. crabs were collected from local market. Spermathecaes five female cut open, contents spermatophore transferred into Ca 2+ free seawater pooled single tube. was gently homogenized by using glass tissue grinder. debris removed nylon mesh filter sperm, suspended seawater, pelleted centrifugation at 400μg 3～5 min then resuspended fresh seawater. A stock solution A23187 dissolved DMSO diluted concentration 2mg/ml optimised period assayed ionophore induction. Spermatozoa exposed 0 (control group) 50 μg/ml 10, 20, 30, 40, 50, 60 120 fixed 3% glutaraldehyde. Three factors levels including series pH (6 0, 7 8 9 10 0), different concentrations (32, 48, 56 64 μg/ml) (0 10%, 15%, 20%, 25% 30%) chosen assay condition mud crab an orthogonal experiment. After incubation various test solutions 40 min, spermatozoa adding equal volume buffered 6% glutaraldehyde, scored light microscopy. Each score involved random examination 200 spermatozoa. Some samples post 1% O SO 4, dehydrated acetone, embedded epoxy resin, knives, stained uranyl acetate lead citrate, viewed under transmission electron microscope (JEM CXII) scanning (S 520) observe fine structural events reaction. results showed that most Mud completed minutes (Table 1). experiment shown table 1, analysed 3. Comparing R value each column 3, found largest value, 75 6 derived pH. T therefore, (A) effect factor, second factor (C, =31 8), third one (B, =26 2). highest achieved when solutions. Both higher lower suppressed success According K combination 4B 4C 5. In rate. re proceeded 2C 5, counted treatment. 73 5% 82 5 respectively. It concluded (82 5%) can be isolated spermathecae are A23187, 40min.