Lipoprotein mediated lipid uptake in oocytes of polychaetes (Annelida)

Lipoprotein mediated lipid uptake in oocytes of polychaetes (Annelida)
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脂蛋白介导的多毛类卵母细胞的脂质摄取(环节动物)

DOI:
10.1007/s00441-009-0817-7
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发表时间:
2009
影响因子:
3.6
通讯作者:
U. Hoeger
U. Hoeger
中科院分区:
生物学3区
文献类型:
--
作者:
S. Schenk;U. Hoeger

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本文研究了1,1‘-dioctadecyl-3,3,3’,3‘-tetramethylindocarbocyanine高氯酸盐(DiI)标记的性别非特异性核糖脂蛋白在多毛虫卵母细胞中的摄取。荧光标记首先在内吞泡中观察到(直径1μm),然后融合成更大的小泡(2-3μm);这些小泡最后被结合到现有的未标记的卵黄颗粒(5-6μm)中。在同卵母细胞中,发育末期的卵母细胞与发育早期的卵母细胞相比,内吞泡的融合延迟。用异硫氰酸荧光素(FITC)和DiI双标记的脂蛋白显示,在掺入卵黄颗粒的过程中,蛋白质和脂类部分仍然是共定位的。未观察到胞浆脂滴的标记。客栈。病毒,未标记的核糖脂蛋白作为DiI标记的核糖脂蛋白的竞争性抑制剂是有效的。配基印迹实验证实了脂蛋白受体的存在,其表观分子质量为120 kDa,与已知的卵黄蛋白受体不同。这表明,在多毛类卵母细胞中,存在两种不同的受体,分别介导卵黄蛋白和脂蛋白的摄取。因此,性别无关的脂蛋白除了摄取已知的卵黄蛋白相关的脂类外,还对发育中的卵母细胞的脂类供应做出了贡献。即使在长时间与标记的脂蛋白孵育后,细胞质中的脂滴中也没有标记,这表明这些脂类是通过脂蛋白衍生的脂类的分解和重新合成和/或卵母细胞内的Byde新合成而产生的。
The uptake of the 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled sex-unspecificNereislipoprotein was investigated in oocytes of the nereidid polychaetesNereis virensandPlatynereis dumerilii. The fluorescence label was first observed in endocytic vesicles (<1 μm diameter), which later fused to larger vesicles (2–3 μm); these were finally incorporated into existing unlabeled yolk granules (5–6 μm). InPlatynereisoocytes, 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. InN. virens, unlabeledNereislipoprotein was effective as a competitive inhibitor of DiI-labeledNereislipoprotein. 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 byde novosynthesis within the oocyte.
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