title: Assembly of nuclear pore complexes and annulate lamellae promotes normal pronuclear development in fertilized mammalian oocytes.
abstract: In addition to functional nuclear pore complexes engaged in nucleo-cytoplasmic transport, the cytoplasmic stacks of pore complexes, called annulate lamellae, exist in numerous cell types. Although both annulate lamellae and nuclear pore complexes are present in fertilized mammalian oocytes, their relative roles in the process of fertilization and preimplantation development are not known. Using epifluorescence and electron microscopy, we explored their fate during bovine fertilization. The assembly of annulate lamellae in bovine oocytes was triggered by sperm-oocyte binding and continued concomitantly with the incorporation of the nuclear pores in the nuclear envelopes of the developing male and female pronuclei. This process was also induced by the parthenogenetic activation of metaphase-II-arrested oocytes. Depletion of Ca2+, previously implicated in oocyte activation and in the insertion of pore complexes into the nuclear envelope, prevented the formation of nuclear pore complexes, but not the assembly of annulate lamellae in oocyte cytoplasm. Injection of the nuclear pore antagonist, wheat germ agglutinin, into the cytoplasm of mature oocytes that were subsequently fertilized caused the arrest of pronuclear development, indicating the requirement of nuclear pore complexes for normal pronuclear development. Treatment of the fertilized oocytes with the microtubule inhibitor, nocodazole, prevented gathering of annulate lamellae around the developing pronuclei, insertion of nuclear pores into their nuclear envelopes, and further pronuclear development. The formation of the male pronuclei was reconstituted in Xenopus egg extracts and reflected the behavior of nuclear pores during natural fertilization. These data suggest that nuclear pore complexes are required for normal pronuclear development from its beginning up until pronuclear apposition. Annulate lamellae may be involved in the turnover of nuclear pore complexes during fertilization, which is in turn facilitated by the reorganization of oocyte microtubules and influx of Ca2+ into oocyte cytoplasm.
authors.affiliation: Departments of Obstetrics and Gynecology, and Cell and Developmental Biology, Oregon Health Sciences University, Portland, Oregon, USA.
authors.forename: P
authors.initials: P
authors.lastname: Sutovsky
authors.forename: C
authors.initials: C
authors.lastname: Simerly
authors.forename: L
authors.initials: L
authors.lastname: Hewitson
authors.forename: G
authors.initials: G
authors.lastname: Schatten
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D005260:Female; D005306:Fertilization / Q000502:physiology; D066298:In Vitro Techniques; D008297:Male; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000187:drug effects; D015739:Nocodazole / Q000494:pharmacology; D009685:Nuclear Envelope / Q000187:drug effects / Q000648:ultrastructure*; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D010312:Parthenogenesis; D014909:Wheat Germ Agglutinins / Q000494:pharmacology; D014981:Xenopus; D015053:Zygote / Q000254:growth & development* / Q000378:metabolism / Q000648:ultrastructure*
chemical_list: D014909:Wheat Germ Agglutinins; D015739:Nocodazole; D002118:Calcium