1 / 18

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

SEA URCHIN EMBRYO - A PROMISING MODEL IN SCREENING FOR TUBULIN DESTABILIZING ACTIVITY. a. b. MICROTUBULES. MICROTUBULES. HeLa. 10-15 pf. INTRACELLULAR TRANSPORT. MITOSIS. CELL SHAPE AND MIGRATION. CILIA & FLAGELLA. THE ROLES OF MICROTUBULES IN CELL. FUNCTIONS OF TUBULIN/MICROTUBULE.

mattox
Download Presentation

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

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


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

  2. a b MICROTUBULES MICROTUBULES HeLa 10-15 pf

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

  4. DESTABILIZING AGENTS: INHIBIT POLYMERIZATION DISASSEMBLE MICROTUBULES a b STABILIZING AGENTS: ALTER MICROTUBULE DYNAMICS TUBULIN: TUBULIN - TARGET A TARGET FOR ANTI-TUMOR DRUGS HETERODIMER MICROTUBULE • MITOTIC ARREST, ANTI-PROLIFERATIVE ACTIVITY • SHUTDOWN OF TUMOR BLOOD SUPPLY

  5. MICROTUBULE DESTABILIZING DRUGS DESTAB/STRUCTURES 2-METHOXY-ESTRADIOL COLCHICINE PODOFYLLOTOXIN COMBRETASTATIN b D-24851 a NOSTODIONE NOCODAZOLE VINBLASTINE DOLASTATIN

  6. + + b a b Diaz et al., J Biol Chem, 2000 MICROTUBULE STABILIZERS STABILIZERS DISCODERMOLIDE TAXOL EPOTHILONES

  7. NEW ANTI-TUMOR TUBULIN DESTABILIZING DRUGS NEW ANTI-TUB. DRUGS

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

  9. BLASTULA ACTIVE FEEDING SEA URCHIN EMBRYONIC DEVELOPMENT STAGES HATCHING 9-16 h CLEAVAGE MESENCHYME BLASTULA LATE GASTRULA FERTILIZED EGG ~ 115 m 450 - 500 m ~ 140 m PRISM EARLY PLUTEUS MID-PLUTEUS 2-3 DAYS

  10. a b TUBULIN SYNTHESIS DURING SEA URCHIN EMBRYO DEVELOPMENT SU_EMBR. TUBULIN HATCHING IV III I II Gong & Brandhorst, Dev Biol, 1988 (Lytechinus pictus)

  11. NH CROSS-PRECIPITATION WITH ANTIBODIES TO VERTEBRATE BRAIN TUBULIN (CHICKEN, RAT, PIG) HIGHLY CONSERVATIVE POLYPEPTIDE BACKBONE ~ 450 АMINO ACIDS2-17% VARIABILITY VARIABLE DOMAIN ~ 15-17 АA 2 a b COOH SIMILARITY OF TUBULINS FROM SEA URCHIN EMBRYOS AND VERTEBRATES TUBULIN SU & VERT I II III FITC - LABELED PIG BRAIN TUBULIN INCLUDES INTO MITOTIC SPINDLE Y.Hamagushi et al., J Cell Biol, 1985 (Clypeaster japonicus)

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

  13. EFFECT OF ANTI-PROLIFERATIVE AGENTS EFFECT/ANTIPROLIFER. TARGET - DNA AZIDOTHYMIDINE АPIGENIN HYDROXYUREA MITOXANTRONE MITOMYCIN С RUBOXYL 4-PHENYLBUTYRIC ACID 5-FLUOROURACYL APHIDICOLIN 5-AZACYTIDINE DOXORUBICIN CHLORAMBUCIL TARGET - CYTOSKELETON ACRYLAMIDE CYTOCHALASIND ABNORMAL CLEAVAGE NO SPINNING

  14. EFFECTIVE CONCENTRATIONS EFFECTIVE CONC. OF TUBULIN DESTABILIZING DRUGS SEE TUBULIN DESTABILIZERS DATABASE ON PUBCHEM >>>

  15. b a SCREENING PROCEDURE FOR TUBULIN DESTABILIZERS SCREENING METHOD CLEAVAGE ARREST A CLEAVAGE FERTILIZED EGG Б SWIMMING BLASTULA SPINNING

  16. 100mM 10 mM 5мM FERTILIZED EGG 0.05mM 0.2mM 50 mM 10 mM 1 mM Control 5 ml SUSPENSION IN SEA WATER, 700-3000 EMBRYOS/ML ONE FEMALE EMBRYOS  ~ 30 PLATES TREATMENT PROCEDURE TREATMENT PROCEDURE EXPOSED STAGES STOCK SOLUTIONS(IN DMSO, ETHANOL) HATCHED BLASTULA 9 h (20-23о) 16 h (14-17о)

  17. LATE BLASTULA OREGON GREEN TAXOL MICROTUBULE VISUALIZATION OGT-TEST 2005 CONTROL COLCHICINE, NOCODAZOLE, D-24851 3 NEW COMPOUNDS

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

More Related