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14 A pollo shaped malformation 15. exogastrula Fig. 1 @ The main stages of early development of the sea urchin embryo i.

2 14 A pollo shaped malformation 15. exogastrula Fig. The main stages of early development of the sea urchin embryo i. The 8 cell stage s formed in about 2h after fertilization, the swimming blastula abo ut 8h and the pluteus about 24h later. 1. unfertilized egg, 2. fertilized egg, fertilization membrane was el 3 evated.. 2 cell stage, 4. 4 cell stage 5. 8 cell stage 6. morula stage 7. early blastula 8. just hatching blastula 9. swimming blastula 10. gastrula 11. prism shaped pluteus. bonification is biginning 12, 13 pluteus

3 Table Effect of heavy metal ion on the development of Temnopleurus eggs in Herbst's artificial seawater.

5 Table Effect of heavy metal ion on the development of Hemicentrotus egg in filtered sea water.

Table 3 @ Subthreshold concentration of he avy

6 Table Subthreshold concentration of he avy metal ion on the development of sea urchin eggs

7 Table Effect of two metal ion combination on the development of Temnopleurus eggs in filtered seawater.

1) Freeman, H. C.: Metal complexes of amino acids and peptides. In Eichhorn, G. L. ed. Inorganic Biochemistry, Vol. 1. Pp. 121-166, Elsevier Sci. Pub. Co., Amsterdam, 1973. 3) Maniloff, J.

11 1) Freeman, H. C.: Metal complexes of amino acids and peptides. In Eichhorn, G. L. ed. Inorganic Biochemistry, Vol. 1. Pp , Elsevier Sci. Pub. Co., Amsterdam, ) Maniloff, J., Coleman, J. R., and Miller, M. W.: Effects of Metals on Cells., Subcellular Elements, and Macromolecules. Charles C Thomas Pub., Springfield, ) Bryan, G. W.: The effects of heavy metals (other than mercury) on marine and estuarine organisms. Proc. Roy. Soc. Lond. B. 177, , ) Morisawa, M., and Mori, H.: Heavy metals and Spermatozoan motility I. Distribution of iron, zinc and copper in sea urchin spermatozoa. Expt. Cell Res., 70, , ) Thrasher, J. P.: The effect of mercuric compounds on dividing cells. In Zimmerman, A. M., Padilla, G. M., and Cameron, I. L. ed. Drugs and the Cell Cycle. Pp , Academic Press, New Tork, ) Falchuk, K. H., Fawcett, D. W., and Vallee, B. L.: Role of Zinc in Cell division of Euglena gracilis. J. Cell Sci., 17, 57-78, ) Altman, P. L. and Dittmer, D. S.: Biology Data Book, Vol. I. (second ed.) Pp , Federation of American Societies for Experimental Biology, Bethesda, ) Kobayashi, N.: Fertilized sea urchin eggs as an indicatory material for marine pollution bioassay, preliminary experiments. Publ. Seto Mar. Biol. Lab., 18, , ) Kobayashi, N.: Marine pollution bioassay by sea urchin eggs, an attempt to enhance accuracy. Publ. Seto Mar. Biot. Lab., 21, , 1974.

12 The effects of heavy metals on developing sea urchin eggs. Tetuhide H. MURAKAMI, Masashi HAYAKAWA, Toshitake FUJII, Takehito HARA, Yoshiaki ITAMI, Akira KISHIDA, and Isamu NISIDA. (Department of Physiology, Okayama University Medical School) The effects of heavy metals, such as Hg, Pb, Cu, Cd, As, Sn, Cr, Mn, Ni and Zn on the two developing sea urchin eggs. Temnopleurus and Hemicentrotus were invostigated. The results obtained were as follows. 1. The development of fertilized eggs was inhibited when embryos were exposed to 10-6M of HgCl2 and CuCl2 and to 10-4M of CrCl2 and NiCl2 in Herbst's artificial seawater from fertilization until the formation of the blastula larva. 2. Chlorate and acetate of heavy metals were dissolved in filtered seawater. The morpho logical differences of embryo were not observed until the formation of the pluteus larva in the two solutions. Inhibitory effects were observed when embryos were exposed to seawater at concent rations of 107M of Hg and Cu, to 10-6M of Cd, and to 10-5M of Pb, Ni and Zn. At a concent ration of 10-4M of Cd there were no visible effects until the appearance of swimming blastula larva, but larva development was strongly inhibited at time of bonification from pluteus larva, and there was no further later development. In 10-6M Hg, Cr, and Ni, the larvae were structural abnonal and shaped like Apollo. In 10-7M of Hg exogastrula embryos were produced. 3. Threshold concentration of complete inhibition on the early development of sea urchin embryos was at 10-6M of Hg, 10-5M of Cu, Cr, Ni, and Zn, 10-3M of Mn. In the case of Cd 10-5M, complete inhibition didn't occur until the pluteus larva stage. 4. Only additive effects were obtained on the developing sea urchin eggs when two heavy metals were combined in filtered seawater. A high frequency of morphological abnormalities were formed such as exogastrula and Apollo shaped pluteus especially in coupling of Hg and Cu or Cd and Zn. 5. There were no visible effects in filtered sewage from fertilization to the formation of the pluteus larva. When embryos were exposed to acetate salts of Hg, Cu, Cd, and Zn in filtered sewage, there were 10 fold inhibitory effects on development. But formation of skeleton in the pluteus larva was found in filtered sewage containing Cd. 6. Only additive effects were observed on sea urchin embryo when combined with Hg, Cu, Cd, and Zn in filterd sewage. But the abnormalities were decreased in all cases. Inhibition of bonification were reduced in mixture of Cd and filtered sewage.

CHEMOTHERAPY APR. 1984

VOL.32 S-3 CHEMOTHERAPY dihydro-4-oxo-7-(1-piperazinyl)-1, 8-naphthyridine- CHEMOTHERAPY APR. 1984 VOL.32 S-3 CHEMOTHERAPY Table 1 Implantation rates and post- implantation survival rates in females mated