Cationized ferritin and phosvitin uptake by coated vesicles of the early chick embryo

Summary

The endocytosis of cationized ferritin and of a phosvitinferritin conjugate by cells of the chick embryo area pellucida has been examined. Cationized ferritin was bound mainly to the free surface of the epiblast but was absent from the region of the primitive streak. The binding was patchy and experiments suggest that the anionic sites which bind cationized ferritin are themselves naturally clustered. Uptake of cationized ferritin was exclusively by coated pits. The resulting coated vesicles delivered the cationized ferritin to membrane-bound sites of accumulation in the cytoplasm and to the close vicinity of Golgi bodies. The cationized ferritin was frequently found to share intracellular vacuoles with yolk granules. The uptake was not affected by the presence of microfilament or microtubule-inhibiting agents. Native ferritin, even at concentrations forty times that of cationized ferritin, was not bound or endocytosed.

Coated pits in the epiblast were often associated with overlying extracellular yolk granules. This suggested that the yolk might be inducing the formation of the coated pits. The yolk protein phosvitin was coupled to ferritin and this conjugate was found to be endocytosed by coated pits. This uptake was inhibited in the presence of an excess of free phosvitin but not by albumin, indicating some selectivity for phosvitin over other proteins. The phosvitin conjugate was also found sharing intracellular vacuoles with yolk. We conclude that the cells of the area pellucida, and in particular those of the epiblast, have an active coated vesicle uptake system which may be able to selectively endocytose yolk or yolk protein.