. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 352 s. SUZUKI. FIGURE 8. Ouchterlony agar diffusion analysis showing reaction between anti-Vn-4 serum (A) against ovarian homogenate (1 and 2) or hemolymph of females (3-6). 1 and 4, stage C; 2 and 5, stage D; 3, hibernating; 6, stage E. glycolipoproteins, as stained with periodic acid/Schiff reagent and Sudan black B. The ovarian homogenate, prepared from immature oocytes (350-450 j/m), was ana- lyzed by the pore-limited method (PAGGE) using standard proteins to determine the molecular weight of the four vitellins (Fig. 4).
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 352 s. SUZUKI. FIGURE 8. Ouchterlony agar diffusion analysis showing reaction between anti-Vn-4 serum (A) against ovarian homogenate (1 and 2) or hemolymph of females (3-6). 1 and 4, stage C; 2 and 5, stage D; 3, hibernating; 6, stage E. glycolipoproteins, as stained with periodic acid/Schiff reagent and Sudan black B. The ovarian homogenate, prepared from immature oocytes (350-450 j/m), was ana- lyzed by the pore-limited method (PAGGE) using standard proteins to determine the molecular weight of the four vitellins (Fig. 4). They were estimated to be Vn-1- 700,000 (700K), Vn - 2-620,000 (620K), Vn • 3-540,000 (540K), and Vn - 4-470,000 (470K), respectively. Figures 5 and 6 show the polypeptide composition of vitellin separated in SDS- PAGE. Vn-4, the major vitellin in stages C and E (Fig. 2-8, 12), yielded three main polypeptides with molecular weights estimated to be 80,000 (80K), 99,000 (99K), and 127,000 (127K) (Fig. 5). To compare the polypeptide compositions of Vn-1- Vn-4, each vitellin prestained with Sudan black B was isolated from immature oo- cytes using PAGE. After electrophoresis, the gel band of each vitellin was cut out and eluted in Tris-HCl buffer. Although the isolated vitellin overlapped with other vitel- lins (Fig. 6-1-4) each vitellin was analyzed on SDS-PAGE. Vn- l-Vn-4 were sepa- rated into several polypeptides; their main polypeptide compositions were very sim- ilar (Fig. 6 • 5-8). The lower polypeptides (MW 80K and 99K) were common through Vn-1-Vn-4 (Figs. 5,6). Immunochemical reactions of and-Vn-4 serum against ovarian homogenate (vi- tellin) and diluted hemolymph (vitellogenin) were examined by double diffusion analysis in agarose gel. The four vitellogenins gave a single precipitate line when anti- Vn • 4 serum was reacted with the hemolymph of stage D females. The precipitate line was absent when the serum was reacted with hemolymph of males (Fig. 7). Figure 8 pres
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
