Fatty acid alteration and the lateral diffusion of lipids in the plasma membrane of keratinocytes
Fatty acid alteration and the lateral diffusion of lipids in the plasma membrane of keratinocytes
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DOI：
10.1006/excr.1997.3553
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发表时间：
1997-05-25
影响因子：
3.7
通讯作者：
Marcelo, CL
中科院分区：
文献类型：
--
作者：
Fulbright, RM;Axelrod, D;Marcelo, CL
荧光探针diI用于研究四种不同培养基中体外培养的成人活角质形成细胞的脂质横向流动性，一种培养基是必需脂肪酸缺乏（EFAD）和低钙离子，这种培养基已知能产生快速增殖的细胞，并且含有极低水平必需脂肪酸的脂质。另外两种培养基中添加必需脂肪酸（FAS），这种培养基已知会导致细胞生长更慢，并且具有标准化的脂肪酸比例。第四种培养基由1 μ M全反式维甲酸添加到脂肪酸补充培养基（FAS- ra）中，这种培养基已知会产生高度增殖的细胞，其生长速度大于FAS菌株，与EFAD菌株相似。采用光漂白后荧光恢复（FRAP）技术对生长在这四种培养基中的角化细胞进行了研究，测定了diI在质膜中的横向扩散速率，结果表明，生长速率与扩散系数(D)呈正相关：diI在EFAD或FAS- ra细胞中的扩散系数高于FAS细胞。FAS细胞的D测量分为两组，一组类似于EFAD细胞的单一组，但另一组由低得多的D值组成。其他FRAP参数（可移动分数和漂白剂深度）在“慢”组大于“快”组，在“快”组中这些参数与D也呈负相关趋势。这些结果被解释为可能与脂肪酸改变间接相关的膜结构或形态的变化，包括衰老角质形成细胞中可能存在的质膜崩溃形成脂质双层相互作用系统的区域。(C) 1997学术出版社。
The fluorescent probe diI was used to study the lateral mobility of lipids in in vitro strains of Living adult human keratinocytes grown in four different media, One medium was essential fatty acid deficient (EFAD) and low in calcium ion, a medium known to yield cells that proliferate rapidly and contain lipid with extremely low levels of essential fatty acids. Two other media were supplemented with essential fatty acids (FAS), media that are known to result in cells that grow more slowly and have normalized fatty acid proportions, A fourth medium consisted of 1 mu M all-trans-retinoic acid added to the fatty acid-supplemented medium (FAS-RA), a medium known to produce cells that are highly proliferative, with a growth rate greater than that of the FAS strains and similar to that of the EFAD strains, The keratinocytes grown in these four media were studied using the fluorescence recovery after photobleaching (FRAP) technique to determine the lateral diffusion rate of diI in the plasma membranes, Our results showed a positive correlation between growth rate and diffusion coefficient (D): the diffusion coefficient of diI was higher in the EFAD or FAS-RA cells than in the FAS cells. The measurement of D among the FAS cells fell into two groups, One group was similar to the single group seen in the EFAD cells, but the other group was composed of much lower D values. The other FRAP parameters (mobile fraction and bleach depth) were larger in the ''slow'' group than in the ''fast'' group, This trend of negative correlation between these parameters and D was also found within the fast group. These results are interpreted in terms of possible changes in membrane structure or morphology that might be indirectly associated with the fatty acid alterations, including the possible presence of areas in senescing keratinocytes where plasma membranes collapse to form an interacting system of lipid bilayers. (C) 1997 Academic Press.