Abnormal assembly of annulate lamellae and nuclear pore complexes coincides with fertilization arrest at the pronuclear stage of human zygotic development.
BACKGROUND
The assembly of nuclear pore complexes (NPC) and their cytoplasmic stacks, annulate lamellae (AL), promote normal nucleocytoplasmic trafficking and accompany pronuclear development within the mammalian zygote. Previous studies showed that a percentage of human oocytes fertilized in vitro failed to develop normal pronuclei and cleave within 40-48 h post insemination. We hypothesized that an aberrant recruitment of NPC proteins, nucleoporins and/or NPC preassembled into AL, might accompany human fertilization arrest.


RESULTS
We explored NPC and AL assembly in unfertilized human oocytes, and fertilized and arrested zygotes by immunofluorescence with an NPC- and AL-specific antibody, mAb 414, and by transmission electron microscopy. Major NPC or AL assembly was not observed in the unfertilized human oocytes. Once fertilization took place, the formation of AL was observed throughout the cytoplasm and near the developing pronuclei with NPC. On the contrary, NPC assembly was disrupted in the arrested zygotes, whereas AL were clustered into large sheaths. This was accompanied by the lack of NPC incorporation into the nuclear envelopes.


CONCLUSIONS
We conclude that the aberrant assembly of NPC and AL coincides with early developmental failure in humans.
Rawe
V Y
VY
Centro de Estudios en Ginecología y Reproducción, CEGyR, Buenos Aires, Argentina. vrawe@mail.magee.edu
Olmedo
S Brugo
SB
Nodar
F N
FN
Ponzio
R
R
Sutovsky
P
P
D000911:Antibodies, Monoclonal; D003593:Cytoplasm / Q000502:physiology*; D004247:DNA / Q000378:metabolism; D005314:Embryonic and Fetal Development; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008854:Microscopy, Electron; D009685:Nuclear Envelope / Q000502:physiology*; D022022:Nuclear Pore / Q000502:physiology*; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D009865:Oocytes / Q000502:physiology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D028861:Nuclear Pore Complex Proteins; D004247:DNA