Lipoprotein mediated lipid uptake in oocytes of polychaetes (Annelida)

Abstract

The uptake of the 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled sex-unspecific Nereis lipoprotein was investigated in oocytes of the nereidid polychaetes Nereis virens and Platynereis dumerilii. The fluorescence label was first observed in endocytic vesicles (<1 microm diameter), which later fused to larger vesicles (2-3 microm); these were finally incorporated into existing unlabeled yolk granules (5-6 microm). In Platynereis oocytes, the fusion of endocytic vesicles was delayed in oocytes at their final stage of development compared with those at an early stage of development. Lipoprotein double-labeled with fluorescein isothiocyanate (FITC) and DiI revealed that both the protein and the lipid moiety remained co-localized during incorporation into the yolk granules of the oocyte. No labeling of the cytoplasmic lipid droplets was observed. In N. virens, unlabeled Nereis lipoprotein was effective as a competitive inhibitor of DiI-labeled Nereis lipoprotein. Ligand blot experiments demonstrated the presence of a lipoprotein receptor with an apparent molecular mass of 120 kDa, which is different from that of the known yolk protein receptor. This indicates the presence, in the polychaete oocyte, of two distinct receptors mediating yolk protein and lipoprotein uptake, respectively. Thus, the sex-unspecific lipoprotein contributes to the lipid supply of the growing oocyte in addition to the known uptake of the yolk-protein-associated lipids. The absence of label in the cytoplasmic lipid droplets, even after prolonged incubation with labeled lipoprotein, suggests that these lipids arise either by the breakdown and resynthesis of lipoprotein-derived lipids and/or by de novo synthesis within the oocyte.