Nagata I, Keilhauer G, Schachner M
Brain Res. 1986 Jan;389(1-2):217-32. doi: 10.1016/0165-3806(86)90190-2.
To study the cellular heterogeneity of astrocytes from early postnatal mouse cerebellum in culture, Bergmann glia were enriched by hand-dissection of Purkinje, molecular and external granular layers ('outer' layer) and fibrous astrocytes of white matter and deep cerebellar nuclei ('inner' layer). Both populations of GFA protein and vimentin-positive astrocytes express N-CAM and the L2/HNK-1 epitope, but not tetanus toxin receptors or A2B5 antigen, at levels detectable by indirect immunofluorescence procedures. The two astrocyte populations are thus indistinguishable from each other. Expression of tetanus toxin receptors and A2B5 antigen in these astrocytes can, however, be induced by removal of neurons. The expression of tetanus toxin receptors is again reduced by readdition of purified populations of small cerebellar neurons. Morphology and proliferation of astrocytes from both layers is also dependent on the presence of neurons: removal of neurons leads to an epithelioid, rather than star-shaped morphology and a severalfold increase in proliferation. Readdition of neurons induces astrocytes to return to their star-shaped morphology. Epidermal growth factor increases proliferation in both populations of astrocytes. We conclude that neither antigenic marker profile, morphology nor proliferative responses serve to distinguish between enriched Bergmann glia and enriched fibrous astrocytes.
为了研究培养的出生后早期小鼠小脑星形胶质细胞的细胞异质性,通过手工解剖浦肯野细胞层、分子层和外颗粒层(“外层”)以及白质和小脑深部核团(“内层”)的纤维性星形胶质细胞来富集伯格曼胶质细胞。两种胶质纤维酸性蛋白(GFA)和波形蛋白阳性的星形胶质细胞群体均表达神经细胞黏附分子(N-CAM)和L2/HNK-1表位,但通过间接免疫荧光法检测时,不表达破伤风毒素受体或A2B5抗原。因此,这两种星形胶质细胞群体彼此难以区分。然而,在这些星形胶质细胞中,去除神经元可诱导破伤风毒素受体和A2B5抗原的表达。再次添加纯化的小脑小神经元群体可使破伤风毒素受体的表达再次降低。两层星形胶质细胞的形态和增殖也依赖于神经元的存在:去除神经元会导致上皮样形态而非星形形态,并使增殖增加数倍。再次添加神经元可诱导星形胶质细胞恢复其星形形态。表皮生长因子可增加两种星形胶质细胞群体的增殖。我们得出结论,无论是抗原标志物谱、形态还是增殖反应,都无法区分富集的伯格曼胶质细胞和富集的纤维性星形胶质细胞。