. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. AMEBOCYTES IN LIMULUS EMBRYOS 23 iron hematoxylin. Giemsa staining provides easy identifica- tion of different tissues, but the amebocyte granules stain only sporadically. Iron hematoxylin reproducibly stains the amebocyte granules, but the staining is similar in appear- ance to staining of the yolk. Despite these limitations, both staining techniques yielded the same result. Amebocytes were identified in stage 18 and older embryos (Fig. 1A-E), but not in earlier stage animals (not shown). To confirm that the cells identifie
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. AMEBOCYTES IN LIMULUS EMBRYOS 23 iron hematoxylin. Giemsa staining provides easy identifica- tion of different tissues, but the amebocyte granules stain only sporadically. Iron hematoxylin reproducibly stains the amebocyte granules, but the staining is similar in appear- ance to staining of the yolk. Despite these limitations, both staining techniques yielded the same result. Amebocytes were identified in stage 18 and older embryos (Fig. 1A-E), but not in earlier stage animals (not shown). To confirm that the cells identified as amebocytes in the Giemsa and hematoxylin stained specimens are in fact ame- bocytes. embryos at the same stages of development were stained using an anti-coagulogen antiserum. On protein blots, the antiserum gives a strong, specific reaction to coagulogen when diluted 1 (arrowhead in Fig. 2B). In immunohistochemical studies, the antiserum specifically stains the coagulogen granules in amebocytes of trilobite larvae (Fig. 3A, B). In embryos, the antiserum specifically stains cytoplasmic granules in amebocytes beginning at stage 18 (Fig. 3C-E) of embryogenesis. No antibody stain- ing was detected in earlier stage embryos (not shown). In embryos at stages 18 and 19, the amebocytes were typically located in areas devoid of yolk and other ooplas- mic components. These ooplasm-free regions appear to be precursors of (he hemocoel cavities. They are typically located dorsal to the ventral plate of the embryo, between the epidermal cell layer and the layer of squamous meso- dermal cells that overlays the central yolk mass (Fig. I A, B). In some sections there also appears to be a second mesoderm layer directly beneath the epidermal cell layer. Thus the cavities may in fact be forming between two layers of mesodermal cells. In addition to the amebocytes, ele- ments of connective tissue are frequently seen within these cavities (Fig. 1A). In embryos at later stages (late stage
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
