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Receptor-Mediated Chicken Oocyte Growth: Differential Expression of Endophilin Isoforms in Developing Follicles¹

Institute of Medical Biochemistry, Department of Molecular Genetics, BioCenter and University of Vienna, A-1030 Vienna, Austria

Receptor-mediated endocytosis of yolk precursors via clathrin-coated structures is the key mechanism underlying rapid chicken oocyte growth. In defining oocyte-specific components of clathrin-mediated events, we have to date identified oocyte-specific yolk transport receptors, but little is known about the oocytes' supporting endocytic machinery. Important proteins implicated in clathrin-mediated endocytosis and recycling are the endophilins, which thus far have been studied primarily in synaptic vesicle formation; in the present study, as a different highly active endocytic system, we exploit rapidly growing chicken oocytes. Molecular characterization of the chicken endophilins I, II, and III revealed that their mammalian counterparts have been highly conserved. All chicken endophilins interact via their SH3 domain with the avian dynamin and synaptojanin homologues and, thus, share key functional properties of mammalian endophilins. The genes show different expression patterns: As in mammals, expression is low to undetectable in the liver and high in the brain; in ovarian follicles harboring oocytes that are rapidly growing via receptor-mediated endocytosis, levels of endophilins II and III, but not of endophilin I, are high. Immunohistochemical analysis of follicles demonstrated that endophilin II is mainly present in the theca interna but that endophilin III predominates within the oocyte proper. Moreover, in a chicken strain with impaired oocyte growth and absence of egg-laying because of a genetic defect in the receptor for yolk endocytosis, endophilin III is diminished in oocytes, whereas endophilin III levels in the brain and endophilin II localization to theca cells are unaltered. Thus, the present study reveals that the endophilins differentially contribute to oocyte endocytosis and development.

¹ Supported by grants from the Austrian Science Foundation to J.N. and W.J.S. (F-606, F-608, P-13931, and P-13940). The nucleotide sequences have been submitted to the GenBank/EMBL Data Bank with accession numbers AJ439350, AJ439351, and AJ439352.

² Correspondence: Wolfgang Johann Schneider, Institute of Medical Biochemistry, Department of Molecular Genetics, BioCenter and University of Vienna, Dr. Bohr-Gasse 9/2, A-1030 Vienna, Austria. FAX: 43 1 4277 61804; wjs{at}mol.univie.ac.at

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