Cissé S, Schipper H M
Bloomfield Centre for Studies in Aging, Lady Davis Institute for Medical Research, Sir Mortimer B, Davis-Jewish General Hospital, Montreal, Que., Canada.
Brain Res. 1993 Jun 25;615(1):141-6. doi: 10.1016/0006-8993(93)91125-c.
A subpopulation of astrocytes in periventricular regions of aging brain and in cysteamine (CSH)-treated glial cultures contain autofluorescent cytoplasmic granules that exhibit an affinity for Gomori's chrome alum hematoxylin (CAH), and non-enzymatic peroxidase activity. Although shown to be histochemically distinct from lipofuscin, the lack of pure preparations of these glial inclusions has hindered the elucidation of their precise chemical constituents. Using sucrose gradient fractionation and density centrifugation on percoll, we obtained enriched preparations of astrocyte cytoplasmic granules from intact rat brain and CSH-treated astrocyte cultures. The presence and relative purity of these inclusions were confirmed by laser scanning confocal microscopy for red autofluorescent granules, diaminobenzidine histochemistry for non-enzymatic peroxidase activity and chrome alum hematoxylin (Gomori) staining. In the enriched fractions, the smaller granules (0.5-4.0 microns) were spherical and weakly autofluorescent, whereas larger inclusions (5.0-10.0 microns) tended to be intensely autofluorescent and pleomorphic. As in situ, the purified material was argyrophilic and did not stain for lipids. Isolation of these astrocytic inclusions should permit a more thorough characterization of their biochemical contents.
衰老大脑脑室周围区域以及用半胱胺（CSH）处理的神经胶质细胞培养物中的星形胶质细胞亚群，含有自发荧光的细胞质颗粒，这些颗粒对Gomori铬明矾苏木精（CAH）有亲和力，并具有非酶促过氧化物酶活性。尽管已证明这些神经胶质细胞内含物在组织化学上与脂褐素不同，但由于缺乏这些胶质细胞内含物的纯制剂，阻碍了对其精确化学成分的阐明。我们利用蔗糖梯度分级分离法和在Percoll上进行密度离心，从完整大鼠脑和CSH处理的星形胶质细胞培养物中获得了富集的星形胶质细胞细胞质颗粒制剂。通过激光扫描共聚焦显微镜观察红色自发荧光颗粒、二氨基联苯胺组织化学检测非酶促过氧化物酶活性以及铬明矾苏木精（Gomori）染色，证实了这些内含物的存在和相对纯度。在富集级分中，较小的颗粒（0.5 - 4.0微米）呈球形，自发荧光较弱，而较大的内含物（5.0 - 10.0微米）往往自发荧光强烈且形态多样。如同在原位一样，纯化后的物质嗜银，且不着脂质。分离这些星形胶质细胞内含物应能更全面地表征其生化成分。
