Rotstein N P, Aveldaño M I
Instituto de Investigaciones Bioquímicas, Universidad Nacional del Sur, Bahía Blanca, Argentina.
Biochim Biophys Acta. 1987 Sep 25;921(2):221-34. doi: 10.1016/0005-2760(87)90022-1.
(1-14C)-labeled (n-6) eicosatetraenoate, (n-3) docosapentaenoate and (n-3) docosahexaenoate (20:4, 22:5 and 22:6, respectively) are efficiently taken up and actively esterified into the lipids of bovine retina after 2 h incubation. Photoreceptor membranes, mitochondria, microsomes and postmicrosomal supernatants, which display significant differences in phospholipid and fatty acid compositions, are isolated after such incubations to study the labeling of lipids. The lipid classes preferentially labeled with the acids (1) largely differ among and within subcellular fractions, while (2) some common features in the treatment of the three polyenes are observed in each fraction. In all of them, the three acids are actively incorporated in phosphatidylcholine; ethanolamine glycerophospholipid, phosphatidylserine (PS) and phosphatidylinositol (PI) are highly labeled with 22:6, 22:5 and 20:4 respectively; within ethanolamine glycerophospholipid, the three label phosphatidylethanolamine in preference to plasmenylethanolamine. Most of the 14C esterified in mitochondria is in phospholipids. The endoplasmic reticulum produces in addition highly labeled triacylglycerols, also found in cytosol. High levels of 14C-labeled diacylglycerols are observed exclusively in photoreceptor membranes, where the specific radioactivity of PI is very high. The total amounts of 14C incorporated (1) are in general similar within a given fraction for the three polyenes, but (2) largely differ among fractions. The labeling of the highly unsaturated phospholipids of photoreceptor membranes is the lowest, while the postmicrosomal supernatant (whose lipids are relatively the poorest in polyenoic fatty acids) contains most of the labeled lipids isolated from retinas under these conditions. The results indicate that polyunsaturated species of retina phospholipids undergo an active synthesis and turnover, as well as an intense intracellular traffic among membranes.
(1-14C)标记的(n-6)二十碳四烯酸、(n-3)二十二碳五烯酸和(n-3)二十二碳六烯酸(分别为20:4、22:5和22:6)在孵育2小时后能有效地被牛视网膜摄取并积极酯化为脂质。孵育后分离出在磷脂和脂肪酸组成上有显著差异的光感受器膜、线粒体、微粒体和微粒体后上清液,以研究脂质的标记情况。用这些酸优先标记的脂质类别(1)在亚细胞组分之间和组分内部有很大差异,而(2)在每个组分中观察到三种多烯在处理上有一些共同特征。在所有这些组分中,三种酸都能积极掺入磷脂酰胆碱;乙醇胺甘油磷脂、磷脂酰丝氨酸(PS)和磷脂酰肌醇(PI)分别被22:6、22:5和20:4高度标记;在乙醇胺甘油磷脂中,三种酸优先标记磷脂酰乙醇胺而非缩醛磷脂酰乙醇胺。在线粒体中酯化的大部分14C存在于磷脂中。内质网还产生高度标记的三酰甘油,在胞质溶胶中也有发现。仅在光感受器膜中观察到高水平的14C标记二酰甘油,其中PI的比放射性非常高。掺入的14C总量(1)在给定组分内对于三种多烯通常相似,但(2)在组分之间有很大差异。光感受器膜的高度不饱和磷脂的标记最低,而微粒体后上清液(其脂质中多烯脂肪酸相对最少)在这些条件下含有从视网膜分离出的大部分标记脂质。结果表明,视网膜磷脂的多不饱和种类经历活跃的合成和周转,以及膜之间强烈的细胞内运输。