Shimomura K, Yahara S, Kishimoto Y, Benjamins J A
Biochim Biophys Acta. 1984 Sep 12;795(2):265-70. doi: 10.1016/0005-2760(84)90074-2.
Previous studies on myelinating rat brain indicated that microsomes, Golgi-enriched and cytosol fractions may process galactolipids destined for myelin. To extend these findings we labeled brain galactolipids in vivo and determined the specific radioactivity of cerebrosides and sulfatides in several subcellular fractions. 17-day-old rats were treated by intracranial injection with [14C]galactose 60 min prior to and [3H]galactose 15 min prior to killing. Subcellular fractions were prepared from brain stem, and concentrations of cerebrosides and sulfatides were determined, their radioactivity measured and the 3H/14C ratio compared. Our results showed that the heavier Golgi-enriched fraction (designated Fraction 2) is unique in its low galactolipid content and high specific radioactivities of cerebrosides and sulfatides. The low ratio of the specific activity of cerebroside to that of sulfatide in Fraction 2 compared to other fractions indicates that it may be the site of most rapid conversion of newly synthesized cerebrosides to sulfatides. The specific radioactivities of cerebrosides and sulfatides in cytosol are intermediate between those in Golgi-enriched Fraction 2 and microsomes and those in myelin, consistent with the role postulated for cytoplasmic elements in the transport of cerebrosides and sulfatides to myelin.
以往对大鼠脑髓鞘形成的研究表明,微粒体、富含高尔基体的组分和胞质溶胶组分可能参与处理用于髓鞘的半乳糖脂。为了扩展这些发现,我们在体内标记了脑半乳糖脂,并测定了几种亚细胞组分中脑苷脂和硫脂的比放射性。在处死前60分钟经颅内注射[14C]半乳糖、处死前15分钟经颅内注射[3H]半乳糖来处理17日龄大鼠。从脑干制备亚细胞组分,测定脑苷脂和硫脂的浓度,测量其放射性,并比较3H/14C比值。我们的结果表明,较重的富含高尔基体的组分(指定为组分2)的独特之处在于其半乳糖脂含量低,脑苷脂和硫脂的比放射性高。与其他组分相比,组分2中脑苷脂与硫脂比活性的低比值表明,它可能是新合成的脑苷脂最快转化为硫脂的部位。胞质溶胶中脑苷脂和硫脂的比放射性介于富含高尔基体的组分2和微粒体以及髓鞘中的比放射性之间,这与推测的胞质成分在脑苷脂和硫脂向髓鞘转运中的作用一致。