Fig. 1. Time course of 3H-leucine incorporation into the acid-insoluble fraction of the Bombyx egg. Egg: 24hr after oviposition.

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1 Changes in Protein Synthesis during Embryonic Development of the Silkworm, Bombyx mori (Lepidoptera: Bombycidae). Yutaka KAWAGUCHI and Hiroshi FUJII (Laboratory of Sericulture, Department of Agriculture, Kyushu University, Hakozaki, Higashi-ku, Fukuoka 812, Japan). Ja p. J. appl. Ent. Zool. 28: (1984) Protein synthesis in developing embryos of Bombyx mori was studied by means of microinjection experimens using tritium-labeled precursors and actinomycin D (AD). The incorporation of 3Hleucine into the acid-insoluble fraction was shown to be most active on the first day (stages 3-4) and during the fifth to ninth days (stages 6-20) after oviposition. In the diapausing egg the incorporation of 3H-leucine during stages 3 to 4 was not affected by AD, while the incorporation at the later stages was appreciably inhibited by AD. Two-dimensional gel electrophoresis followed by autoradiography using 35S-methionine demonstrated the existence of stage-specific peptides which were synthesized at different phases of development. The present results suggest that the translational event in the early stages of embryogenesis depends on pre-existing maternal mrnas, and at the later stages of the embryonic development it depends upon newly synthesized mrnas.

2 Fig. 1. Time course of 3H-leucine incorporation into the acid-insoluble fraction of the Bombyx egg. Egg: 24hr after oviposition.

3 Fig. 2. Changes of incorporation of 3H-leucine into the acid-insoluble fraction of the Bombyx egg throughout the embryogenesis. Eggs that were incubated at 25 Ž were collected six hours after injection. Artificial hatching treatment treated) (HCl was performed at twenty hours oviposition. œ: after Fig. 3. Effect of actinomycin D at varying doses on the incorporarion of 3H-leucine into the acid-insoluble fraction of the Bombyx egg. Five days old eggs were used. HCl-treated, and non-diapausing egg, : diapausing egg in which HCl treatment was omitted. Fig. 5. Effect of actinomycin D on incorporation of 3Hleucine into the acid-insoluble fraction in the Fig. 4. Time course of 3H-leucine incorporation (2 ~10-10ƒÊg/egg) into the acid-insoluble Bombyx egg throughout the embryogenesis. Eggs were collected six hours after injection. œ: fraction of the Bombyx egg in the presence of actinomycin D. The age of the egg 3H-leucine, : 3H-leucine+actinomycin D. was the same as in Fig. 3.

Fig. 6. Autoradiographs of the two-dimensional gel electrophoresis of 35S-methionine labeled protein. Eggs were allowed a three-hour incorporation of the labeled precursor.

4 Fig. 6. Autoradiographs of the two-dimensional gel electrophoresis of 35S-methionine labeled protein. Eggs were allowed a three-hour incorporation of the labeled precursor. 6th, 1d, 3d and 7d refer to six hours, one-, three- and seven-day old eggs, respectively. One- to seven-day old eggs were previously subjected to the artificial hatching treatment at twenty hours after oviposition.

73 DAVIDOSON, E.H. (1976) Gene activity in early development. 2nd ed. New York: Academic Press, p. 452. GOLDSTEIN, E.S. and L.A. SNYDER (1973) Protein synthesis and distribution of ribosomal elements in ovarian oocytes and early developmental stages of Drosophila melano- gaster.

6 73 DAVIDOSON, E.H. (1976) Gene activity in early development. 2nd ed. New York: Academic Press, p GOLDSTEIN, E.S. and L.A. SNYDER (1973) Protein synthesis and distribution of ribosomal elements in ovarian oocytes and early developmental stages of Drosophila melano- gaster. Exp. Cell Res. 81: GRZELAK, K., E. SZCZESNA and Z. LASSOTA (1979) Polyadenylated RNA in diapausing and developing embryos of Bombyx mori. Insect Biochem. 9: JACKLE, H. (1979) Degradation of maternal poly (A)-containing RNA during early embryogenesis of an insect (Smittia spec. Chironomidae, Diptera). Wilhelm Roux's Arch. Entwicklungsmech. Org. 187: JACKLE, H. and K. KALTHOFF (1979) RNA and protein synthesis in developing embryos of Smittia spec. (Chironomidae, Diptera). Wilhelm Roux's Arch. Entwicklungsmech. Org. 187: KAWAGUCHI, Y. and H. DOIRA (1974) Incorporation and synthesis of protein by the ovaries of Bombyx mori. J. Fac. Agr., Kyushu Univ. 18: LOVETT, J.A. and E.S. GOLDSTEIN (1977) The cytoplasmic distribution and characterization of poly(a)+rna in oocytes and embryos of Drosophila. Dev. Biol. 60: MERMOD, J.J., G. SCHATZ and M. GRIPPA (1980) Specific control of messenger translation in Drosophila oocytes and embryos. Dev. Biol. 75: O'FARRELL, P.H. (1975) High resolution two dimensional electrophoresis of protein. J. Biol. Chem. 250:

7 RAFF, R.A. (1980) Masked messenger RNA and the regulation of protein synthesis in egg and embryos. In: Cell Biology. Vol.4, New York: Academic Press, pp SCHMIDT, O. and H. JACKLE (1978) RNA synthesis during oogenesis and early embryogenesis in an insect egg (Euscolis plebejus). Wilhelm Roux's Arch. Entwicklungsmech. Org. 184: SZYSZKO, M. and Z. LASSOTA (1977) Polysomes in diapausing and developing embryos of Bombyx mori. Insect

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