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博氏沼虾(甲壳纲:长臂虾科)胚胎发育过程中胚胎脂蛋白和卵黄脂磷蛋白的消耗

Embryo lipoproteins and yolk lipovitellin consumption during embryogenesis in Macrobrachium borellii (Crustacea: Palaemonidae).

作者信息

García F, Cunningham M L, Garda H, Heras H

机构信息

Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), CONICET - UNLP, Calles 60 y 120, (1900) La Plata, Argentina.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2008 Nov;151(3):317-22. doi: 10.1016/j.cbpb.2008.07.015. Epub 2008 Aug 5.

Abstract

The prawn Macrobrachium borellii has lecithotrophic eggs with highly-abbreviated development. The major yolk component is lipovitellin (LV), a lipoprotein with 30% lipids (by weight). LV consumption during embryogenesis was followed by ELISA and Western blot analysis using an anti-LV polyclonal antibody. No cross-reacting proteins were observed and LV-like lipoproteins were strongly recognized by the antibody in hemolymph (vitellogenin), yolk (LV) and embryos (LVe), as determined by Western Blot analysis. LV decreased significantly along development from 9.4 to 1.1 microg/mg egg. Consumption rate of LV was slow in early embryogenesis, followed by a rapid utilization in late embryonic stages. Significant LVe amounts were still present at hatching. LV apolipoproteins were selectively degraded during embryo development, being the highest molecular weight subunit the most affected. Comparison among in vitro, in vivo and theoretical proteolysis suggested that trypsin may be involved in LV degradation during late embryogenesis. Embryo lipoprotein (HDLe) synthesis was first detected at stage 6. HDLe shared the same density, MW and subunit composition as adult hemolymph HDL(1) and did not cross-react with LV-like lipoproteins. Though expressed at low concentration, it fulfilled embryo needs for lipid transport among organs.

摘要

博氏沼虾的卵为卵黄营养型,发育高度简化。主要的卵黄成分是卵黄脂磷蛋白(LV),一种脂质含量为30%(重量)的脂蛋白。在胚胎发育过程中,通过酶联免疫吸附测定(ELISA)和使用抗LV多克隆抗体的蛋白质印迹分析来追踪LV的消耗情况。未观察到交叉反应蛋白,通过蛋白质印迹分析确定,该抗体在血淋巴(卵黄蛋白原)、卵黄(LV)和胚胎(LVe)中能强烈识别LV样脂蛋白。随着发育进程,LV显著减少,从9.4微克/毫克卵降至1.1微克/毫克卵。在胚胎发育早期,LV的消耗速率较慢,随后在胚胎后期迅速被利用。在孵化时仍存在大量的LVe。在胚胎发育过程中,LV载脂蛋白被选择性降解,其中分子量最大的亚基受影响最大。体外、体内和理论上的蛋白水解比较表明,胰蛋白酶可能参与胚胎后期LV的降解。胚胎脂蛋白(HDLe)在第6阶段首次被检测到。HDLe与成年血淋巴HDL(1)具有相同的密度、分子量和亚基组成,且不与LV样脂蛋白发生交叉反应。尽管其表达浓度较低,但满足了胚胎在器官间进行脂质运输的需求。

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