Rearrangements of sea urchin egg cytoplasmic membrane domains at fertilization.
Fertilization in the sea urchin is accompanied by rapid reorganization of the egg endoplasmic reticulum (ER). ER-derived vesicles contribute to one of three classes of membranes used in assembling the male pronuclear envelope in vitro. We provide here biochemical evidence for the rearrangement of sea urchin egg cytoplasmic membrane domains at fertilization up to the first mitosis, with respect to two nuclear envelope markers, lamin B and lamin B receptor (LBR), using purified vesicles prepared from homogenates fractionated by floatation on sucrose gradients. In unfertilized eggs, immunoprecipitation data indicate that most of lamin B and LBR are localized in the same vesicles but do not interact. By 3 min post-fertilization, both proteins are more widely distributed across the gradients and by 12 min most of lamin B and LBR are localized in vesicles of different densities. This partitioning is maintained throughout S phase. At mitosis, most lamin B and LBR remain in distinct vesicles, while a small proportion of lamin B and LBR, likely derived from the disassembled nuclear envelope, associate in a minor subset of vesicles. The results illustrate a dynamic reorganization of egg cytoplasmic membranes at fertilization, and the establishment of distinct membrane domains enriched in specific nuclear envelope markers during the first cell cycle of sea urchin development. Additionally, we demonstrate that male pro-nuclear membrane assembly occurs only when both cytosol and membranes originate from fertilized but not unfertilized eggs, suggesting that fertilization-induced membrane rearrangements contribute to the ability of the egg to assemble the male pronuclear envelope.
Collas
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P
Institute of Medical Biochemistry, University of Oslo, Norway. philippe.collas@basalmed.uio.no
Barona
T
T
Poccia
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DL
D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003600:Cytosol; D005260:Female; D005306:Fertilization / Q000502:physiology*; D034921:Lamin Type B; D034882:Lamins; D008297:Male; D008938:Mitosis / Q000502:physiology; D009685:Nuclear Envelope / Q000502:physiology; D009687:Nuclear Proteins / Q000378:metabolism; D010063:Ovum / Q000378:metabolism / Q000502:physiology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D012617:Sea Urchins; D000097682:Lamin B Receptor
D034921:Lamin Type B; D034882:Lamins; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D000097682:Lamin B Receptor