. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. SHRIMP CORTICAL GRANULE cDNA AND ANTIBODY 347 1234 1234 MrXlO-3 205- 116- 97- 66- • I ! B Figure 3. Pcnacits vannamei polypeptides were separated by SDS- PAGE, and stained with Coomassie Blue R (Panel A), or blotted onto nitrocellulose and incubated with a polyclonal antibody to the geneti- cally engineered fusion polypeptide (Panel B). Samples were from an animal with mid-cycle ovaries (oocyte diameter ~ 150 ^m). Lane 1, ovary: 2, hemolymph; 3, muscle; 4, hepatopancreas. Positions of mo- lecular weight markers are indicated
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. SHRIMP CORTICAL GRANULE cDNA AND ANTIBODY 347 1234 1234 MrXlO-3 205- 116- 97- 66- • I ! B Figure 3. Pcnacits vannamei polypeptides were separated by SDS- PAGE, and stained with Coomassie Blue R (Panel A), or blotted onto nitrocellulose and incubated with a polyclonal antibody to the geneti- cally engineered fusion polypeptide (Panel B). Samples were from an animal with mid-cycle ovaries (oocyte diameter ~ 150 ^m). Lane 1, ovary: 2, hemolymph; 3, muscle; 4, hepatopancreas. Positions of mo- lecular weight markers are indicated at left. ules/cell, comprising ~ 10% of the oocyte volume. These estimates are based on a cell diameter of ~226 ^m (Ta- ble I). This cell diameter measurement was an underesti- mate of that in fresh tissue as a result of (a) cell shrinkage (from 320 ^m, see below) during sample preparation and (b) imprecision in determining the cell center due to vari- ations in cell morphology. Developmental profile oft he 200 kDa ovarian polypeptide The contribution of the 200 kDa polypeptide to total protein was determined by ELISA during both ovarian development and embryonic development using the pu- rified fusion protein as the standard. The 200 kDa poly- peptide was not detected in a previtellogenic ovary (oo- cyte diameter = 0) (Fig. 5). It represented 4-5% of the total ovarian protein at oocyte diameter 200-240 fim and ~ 11% of the protein in a fully developed ovary (320 pm). In newly spawned eggs, the 200 kDa polypeptide comprised 3-4% of the protein. By 2 h after the spawn, the 200 kDa polypeptide had declined to 4 h spawn. By 6 h after spawning, the first antennal, second antennal, and mandibular primor- dia were visible, and hatching occurred 16-18 h after spawning. Discussion Cortical granules are found widely in vertebrates and invertebrates; however, the size, abundance, and compo- sition of these granules varies among species (see Guraya, 1982, for review). As shown by light
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
