1. SEA URCHIN EMBRYO - A PROMISING MODEL IN SCREENING FOR TUBULIN DESTABILIZING ACTIVITY

2. MICROTUBULES  HeLa 10-15 pf

3. FUNCTIONS OF TUBULIN/MICROTUBULE MT-ROLES INTRACELLULAR TRANSPORT CELL SHAPE AND MIGRATION MITOSIS CILIA & FLAGELLA

4. TUBULIN - TARGET  MICROTUBULE HETERODIMER DESTABILIZING AGENTS: INHIBIT POLYMERIZATION DISASSEMBLE MICROTUBULES STABILIZING AGENTS: ALTER MICROTUBULE DYNAMICS MITOTIC ARREST, ANTI-PROLIFERATIVE ACTIVITY SHUTDOWN OF TUMOR BLOOD SUPPLY

5. DESTAB/STRUCTURES   COLCHICINE PODOFYLLOTOXIN COMBRETASTATIN D-24851 NOCODAZOLE DOLASTATIN VINBLASTINE NOSTODIONE 2-METHOXY- ESTRADIOL

6. STABILIZERS +    + Diaz et al., J Biol Chem, 2000 TAXOL DISCODERMOLIDE EPOTHILONES

7. NEW ANTI-TUB. DRUGS

8. 0.5 M KCl SPAWNING & FERT. Paracentrotus lividus FERTILIZATION ~ 1 000 000 EGGS/FEMALE

9. STAGES PRISMEARLY PLUTEUS ACTIVE FEEDING CLEAVAGEMESENCHYME BLASTULA MID-PLUTEUS 2-3 DAYS 450 - 500  LATE GASTRULA ~ 140  ~ 115  FERTILIZED EGG HATCHING 9-16 h BLASTULA

10. SU_EMBR. TUBULIN Gong & Brandhorst, Dev Biol, 1988 (Lytechinus pictus) I II III IV  HATCHING

11. TUBULIN SU & VERT HIGHLY CONSERVATIVE POLYPEPTIDE BACKBONE ~ 450 АMINO ACIDS 2-17% VARIABILITY VARIABLE DOMAIN ~ 15-17 АA COOH NH 2  CROSS-PRECIPITATION WITH ANTIBODIES TO VERTEBRATE BRAIN TUBULIN (CHICKEN, RAT, PIG) FITC - LABELED PIG BRAIN TUBULIN INCLUDES INTO MITOTIC SPINDLE Y.Hamagushi et al., J Cell Biol, 1985 (Clypeaster japonicus)

12. EFFECT/TUB. DESTAB. COLCHICINE NOCODAZOLE VINBLASTINE DOLASTATIN 15 D-24851 Control 6-11 h 1-30 h TAXOL

13. EFFECT/ANTIPROLIF ER. TARGET - DNA AZIDOTHYMIDINE АPIGENIN HYDROXYUREA MITOXANTRONE MITOMYCIN С RUBOXYL 4-PHENYLBUTYRIC ACID 5-FLUOROURACYL APHIDICOLIN 5-AZACYTIDINE DOXORUBICIN CHLORAMBUCIL TARGET - CYTOSKELETON ACRYLAMIDE CYTOCHALASIN D ABNORMAL CLEAVAGE NO SPINNING

14. EFFECTIVE CONC. SEE TUBULIN DESTABILIZERS DATABASE ON PUBCHEM >>>

15. SCREENING METHOD  a FERTILIZED EGG CLEAVAGE ARREST SWIMMING BLASTULA SPINNING CLEAVAGE

16. TREATMENT PROCEDURE EXPOSED STAGES HATCHED BLASTULA 50  M Control 10  M 0.05  M0.2  M1  M 5 ml SUSPENSION IN SEA WATER, 700-3000 EMBRYOS/ML ONE FEMALE EMBRYOS  ~ 30 PLATES 9 h (20-23 о ) 16 h (14-17 о ) FERTILIZED EGG 5 мM5 мM 100  M 10  M STOCK SOLUTIONS (IN DMSO, ETHANOL)

17. OGT-TEST 2005 LATE BLASTULA CONTROL COLCHICINE, NOCODAZOLE, D-24851 3 NEW COMPOUNDS

18. EXPENSES 350 - 400 $ IN VITRO TUBULIN POLYMERIZATION 2500 - 10 000 $ IN VIVO TEST ~ 200 $ SEA URCHIN EMBRYO TEST 750 $ CELL CULTURE TEST IC50, FACS, D10