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Lipid species affect morphology of endoplasmic reticulum: a sea urchin oocyte model of reversible manipulation

机译:脂质物种影响内质网的形态:可逆操作的海胆卵母细胞模型

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摘要

The ER is a large multifunctional organelle of eukaryotic cells. Malfunction of the ER in various disease states, such as atherosclerosis, diabetes, cancer, Alzheimer’s and Parkinson’s and amyotrophic lateral sclerosis, often correlates with alterations in its morphology. The ER exhibits regionally variable membrane morphology that includes, at the extremes, large relatively flat surfaces and interconnected tubular structures highly curved in cross-section. ER morphology is controlled by shaping proteins that associate with membrane lipids. To investigate the role of these lipids, we developed a sea urchin oocyte model, a relatively quiescent cell in which the ER consists mostly of tubules. We altered levels of endogenous diacylglycerol (DAG), phosphatidylethanolamine (PtdEth), and phosphatidylcholine by microinjection of enzymes or lipid delivery by liposomes and evaluated shape changes with 2D and 3D confocal imaging and 3D electron microscopy. Decreases and increases in the levels of lipids such as DAG or PtdEth characterized by negative spontaneous curvature correlated with conversion to sheet structures or tubules, respectively. The effects of endogenous alterations of DAG were reversible upon exogenous delivery of lipids of negative spontaneous curvature. These data suggest that proteins require threshold amounts of such lipids and that localized deficiencies of the lipids could contribute to alterations of ER morphology. The oocyte modeling system should be beneficial to studies directed at understanding requirements of lipid species in interactions leading to alterations of organelle shaping.
机译:ER是真核细胞的大型多功能细胞器。 ER在各种疾病状态下的功能异常,例如动脉粥样硬化,糖尿病,癌症,阿兹海默氏症和帕金森氏症以及肌萎缩性侧索硬化症,通常与其形态改变有关。 ER表现出区域可变的膜形态,在极端情况下包括较大的相对平坦的表面和相互连接的管状结构,其横截面高度弯曲。 ER形态是通过塑造与膜脂结合的蛋白质来控制的。为了研究这些脂质的作用,我们开发了海胆卵母细胞模型,这是一种相对静止的细胞,其中的ER主要由小管组成。我们通过酶的微注射或脂质体脂质输送来改变内源性二酰基甘油(DAG),磷脂酰乙醇胺(PtdEth)和磷脂酰胆碱的水平,并通过2D和3D共聚焦成像和3D电子显微镜评估了形状变化。降低和增加脂类(例如DAG或PtdEth)的水平,其特征是负自发曲率分别与转化为片状结构或肾小管有关。 DAG的内源性改变的影响在负性自发曲率脂质的外源性传递后是可逆的。这些数据表明蛋白质需要阈值量的此类脂质,并且脂质的局部缺陷可能会导致ER形态的改变。卵母细胞建模系统应有利于旨在理解相互作用中导致细胞器形状改变的脂质种类要求的研究。

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