Membrane anomalies in Huntington's disease fibroblasts.
Membrane anomalies in Huntington's disease fibroblasts.
复制标题
亨廷顿病成纤维细胞的膜异常。
DOI：
10.1111/j.1471-4159.1984.tb00931.x
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发表时间：
1984
影响因子：
4.7
通讯作者：
Roberts,E
中科院分区：
文献类型：
--
作者：
Schroeder,F;Goetz,IE;Roberts,E
从配对、传代次数匹配的培养人成纤维细胞中分离质膜、微粒体和线粒体。这些细胞是从亨廷顿病（HD）受试者的皮肤活检组织和性别和年龄匹配的对照组中获得的。所有成纤维细胞在相同培养基中培养3 - 7代。表面标记酶如Na+，K+-ATP酶的富集表明分离的质膜纯化了10倍。HD成纤维细胞的粗匀浆和分离质膜中Na+，K+-ATP酶的比活性分别比对照成纤维细胞高62%和82%。质膜Na+，K+-ATP酶的比活性与脂质组成和膜结构相关，通过测量荧光探针分子的旋转弛豫时间和极限各向异性来确定。未观察到HD成纤维细胞质膜结构的重大改变。1，6-二苯基-1，3，5-己三烯和反式帕里纳酸的旋转弛豫时间和极限各向异性在HD的质膜、微粒体或线粒体与对照成纤维细胞之间没有显著差异。反式帕里纳酸证明了正常和HD皮肤成纤维细胞的所有三种亚细胞膜组分中流体和固体区域的共存。最后，反式parinaric酸和1，6-二苯基-1，3，5-己三烯在质膜、微粒体和线粒体中的吸光度校正荧光的Arrhenius图中显示出特征性断点。在所有情况下，在20° C和30°C附近观察到类似的断点温度，表明相变。这些断点在HD中未改变。总之，数据不支持HD中主要膜结构缺陷的概念。
Plasma membranes, microsomes, and mitochondria were isolated from paired, passage number matched, cultured human fibroblasts. The cells were obtained from skin biopsies of Huntington's disease (HD) subjects and from sex and age matched controls. All fibroblasts were cultured in identical media for three to seven passages. Enrichment of surface marker enzymes such as Na+,K+‐ATPase indicated a 10‐fold purification of the isolated plasma membrane. The specific activity of Na+,K+‐ATPase was 62 and 82% greater in the crude homogenate and isolated plasma membrane, respectively, of HD fibroblasts than in control fibroblasts. The specific activity of plasma membrane Na+,K+‐ATPase was correlated with lipid composition and with membrane structure as determined by measurement of the rotational relaxation time and limiting anisotropy of fluorescence probe molecules. Major alterations in the structure of the plasma membranes in HD fibroblasts were not noted. The rotational relaxation time and limiting anisotropy of 1,6‐diphenyl‐l,3,5‐hexatriene and oftrans‐parinaric acid were not significantly different between the plasma membrane, microsomes, or mitochondria of HD versus those of control fibroblasts.trans‐Parinaric acid demonstrated the coexistence of fluid and solid domains in all three subcellular membrane fractions of the normal and HD skin fibroblasts. Lastly, bothtrans‐parinaric acid and 1,6‐diphenyl‐1,3,5‐hexatriene displayed characteristic breakpoints in Arrhenius plots of absorbance corrected fluorescence in plasma membranes, microsomes, and mitochondria. In all cases, similar breakpoint temperatures, indicative of phase alterations, were noted near 20° and 30°C. These breakpoints were unaltered in HD. In summary, the data do not support the concept of major membrane structural defects in HD.