Endocytic traffic in trophectoderm and polarised blastomeres of the mouse preimplantation embryo.

Abstract

Endocytosis from apical and basolateral cell membranes of mouse blastocyst trophectoderm was examined morphologically by using unconjugated horseradish peroxidase (HRP, fluid phase marker), cationized ferritin (membrane marker, bound ionically), and protein A-HRP conjugate (membrane marker, identifying antigens recognised by antimouse species serum). The markers were applied in single and double labelling procedures designed to reveal the derivation, sorting site, and fate of all the major endocytic pathways. Endocytosis at the apical surface led to the obligate fusion of labelled elements with prelysosomal endosomes prior to the redistribution of membrane into lysosomal, transcellular, or recycling pathways, and to the passage of internalised fluid into lysosomes only. Complementary routes appear to operate following endocytosis at the basolateral domain. Thus, endocytic trafficking within the trophectoderm, regulated by "sorting" mechanisms localised at the endosome compartment, may be responsible for the maintenance of polarised membrane domains. The polarised transcellular pathway involving obligatory endosome fusion is present in cleavage-stage, peripheral 16-cell blastomeres prior to zonular tight junction formation. Nocodazole treatment to depolymerize microtubules in many cases induced a bypass of the endosomal sorting compartment during transcytosis, indicating that microtubules contribute to the spatial organization of endocytic membrane traffic.