The structure of the areas pellucida and vasculosa of the early chick embryo (stages 11-29) was examined by light, transmission and scanning electron microscopy. The most striking feature of the endodermal cells of these areas is the presence of large intracellular yolk drops which are characteristic of the regions in which they are found; lipid-like homogeneous drops in the area pellucida, heterogeneously composed pleomorphic drops in the mid-region of the area vasculosa and granular drops at the periphery of the area vasculosa in the region of the sinus terminalis. On morphological criteria it is postulated that granular drops may arise by direct engulfment of extracellular yolk, but this does not appear to be true for pleomorphic or homogeneous drops. Since the apical junctions between endodermal cells across the yolk sac are tight, they seal off the extraembryonic compartment from the vitelline circulation and presumably prevent intercellular passage of the yolk constituents. Thus the endodermal epithelium must mediate the transport of nutrients from the yolk mass to the developing embryo. Endodermal cells exhibit a variation across the yolk sac in the presence and number of structures associated with uptake of extracellular materials. The mid-region of the area vasculosa appears to be the most endocytotically active region with an abundance of microvilli, bristle-coated pits and vesicles and apical canaliculi and vacuoles. There is a close association between the endoderm and vitelline blood vessels and this association is maintained, as the yolk sac develops, by the formation of small vessels juxtaposed between the vascular surface of the endoderm and the walls of the large vitelline vessels.

The endoderm of the chick yolk sac mediates the transfer of materials from the yolk mass to the embryonic circulation. There is little evidence of endocytotic activity in the area pellucida, but the endodermal cells of the area vasculosa possess many microvilli and bristle-coated pits and vesicles, as well as a canalicular system and vacuoles in the apical cytoplasm. Three tracers, horseradish peroxidase, ferritin, and latex spheres, were injected beneath the endoderm of both cultured embryos and embryos in ovo to study the pathway of uptake of extracellular materials. All tracers were sequestered in bristle-coated pits (200-500 nm in diameter) of the endodermal cells of the area vasculosa, but not those in the area pellucida. Both horseradish peroxidase and latex spheres (and probably ferritin) were incorporated into pleomorphic intracellular yolk drops through bristle-coated pits and vesicles, and then into apical vacuoles, which fuse with the intracellular yolk drops. Horseradish peroxidase and ferritin were also found within apical canaliculi. The apical junctions between endodermal cells prevented the intercellular passage of the tracers. A "topping-up" hypothesis is opposed whereby endodermal cells of the area vasculosa continually sequester extracellular yolk material, which is incorporated into a digesting intracellular yolk drop while, at the same time, digested yolk products are being transported across the vascular pole of the endoderm to the extraembryonic circulation and thence to the embryo.

The urodele Ambystoma tigrinum adapts to a distilled water environment by decreasing renal sodium excretion, but the site and mechanism of renal adaptation is unknown. Isolated diluting segments of Ambystoma kidney were studied after a 2-wk exposure of the animals to either distilled or artificial pond ([Na] = 1.2 meq/l) water. Identification of diluting segments was confirmed by electron microscopy. Morphometric study revealed evidence of increased tubular diameter and cellular hypertrophy in the distilled water group. Na+-K+-ATPase activity was increased in tubules from the distilled water group compared with tubules from the pond water group; 40.2 +/- 6.9 vs. 21.7 +/- 4.3 nM ADP generated.min-1.mm tubule length-1, P less than 0.036. This alteration in ATPase activity was due to an increase in the number of pump units present on the basolateral membrane, since specific ouabain binding was also doubled in distilled water compound with pond water group tubules, 20.7 +/- 2.3 vs. 9.1 +/- 0.9 fmol/mm tubule length tubules, P less than 0.011. An increase in transepithelial potential difference of the diluting segment was noted in distilled water group tubules (19.5 +/- 1.4 mV) compared with tubules in the pond water group (13.2 +/- 1.8 mV), P less than 0.015. We conclude that distilled water adaptation is associated with specific diluting segment structural and functional alterations, which are probably linked to an increase in sodium transport rate.

No abstract

The effects of nitrous oxide on the early explanted chick embryo were investigated using the technique of New (1955). After 22 hours incubation the embryos were explanted and exposed for a further 24 hours to the following gases: air, nitrous oxide (25 per cent), nitrous oxide (60 per cent), nitrous oxide (79 per cent). The oxygen concentration was maintained at normal levels (20.9 per cent) and the balance of the pses was nitrogen. In a fifth trial a 60 per cent nitrous oxide 40 per cent oxygen mixture was used. After exposure the embryos were examined under the dissecting microscope and in histological sections. These revealed no effect on the chick embryo after treatment with nitrous oxide for 24 hours.