In vivo analysis of glial cell phenotypes during a viral demyelinating disease in mice.
In vivo analysis of glial cell phenotypes during a viral demyelinating disease in mice.
复制标题
DOI：
10.1083/jcb.109.5.2405
复制
发表时间：
1989-11
期刊：
影响因子：
--
通讯作者：
Dubois-Dalcq M
中科院分区：
文献类型：
--
作者：
Godfraind C;Friedrich VL;Holmes KV;Dubois-Dalcq M
颅内注射小鼠肝炎病毒 A59 株的 C57 BL/6N 小鼠在脊髓神经胶质细胞中表现出广泛的病毒复制，并出现脱髓鞘病变，随后病毒清除和髓鞘再生。为了研究不同的神经胶质细胞类型如何受到疾病过程的影响，我们将三色免疫荧光标记与氚胸苷放射自显影相结合，对 1 微米的脊髓冷冻切片进行观察。我们使用三种不同的神经胶质细胞特异性抗体（a）针对少突胶质细胞表达的 2',3' 环核苷酸 3' 磷酸水解酶（CNP），（b）针对星形胶质细胞表达的神经胶质纤维酸性蛋白（GFAP），以及（c）与大鼠 O-2A 祖细胞结合的 O4 抗体。这些祖细胞产生少突胶质细胞和2型星形胶质细胞，并与成年中枢神经系统中的O4抗体发生反应，在未感染小鼠的脊髓中存在但很少。相比之下，具有 O-2A 祖细胞表型（仅 O4 +）的细胞在病毒接种后一周（1 WPI）数量增加，并且是当时唯一被氚化胸苷体内脉冲 2 小时标记的免疫染色细胞。脊髓中的仅 GFAP+ 和 GFAP+、O4+ 星形胶质细胞在 1 WPI 时也有所增加。在 WPI 的 2 到 4 周之间，受感染的脊髓的特征是脱髓鞘病变内 (CNP+、O4+) 少突胶质细胞的丢失，以及 O-2A 祖细胞和 O4+、GFAP+ 星形胶质细胞的存在，这两种细胞都可以用胸苷标记。随着髓鞘再生的进行，CNP 免疫染色恢复到接近正常，之前在脱髓鞘阶段注射的氚化胸苷现在出现在 CNP+ 少突胶质细胞中。因此，O4 阳性 O-2A 祖细胞在脱髓鞘疾病的早期增殖，而 CNP 阳性少突胶质细胞则不然。事件发生的时间表明，O-2A 祖细胞可能会产生新的少突胶质细胞和 2 型星形胶质细胞，这两种细胞都可能在髓鞘再生过程中发挥作用。
C57 BL/6N mice injected intracranially with the A59 strain of mouse hepatitis virus exhibit extensive viral replication in glial cells of the spinal cord and develop demyelinating lesions followed by virus clearing and remyelination. To study how different glial cell types are affected by the disease process, we combine three-color immunofluorescence labeling with tritiated thymidine autoradiography on 1-micron frozen sections of spinal cord. We use three different glial cell specific antibodies (a) to 2',3' cyclic-nucleotide 3' phosphohydrolase (CNP) expressed by oligodendrocytes, (b) to glial fibrillary acidic protein (GFAP) expressed by astrocytes, and (c) the O4 antibody which binds to O-2A progenitor cells in the rat. These progenitor cells, which give rise to oligodendrocytes and type 2 astrocytes and react with the O4 antibody in the adult central nervous system, were present but rare in the spinal cord of uninfected mice. In contrast, cells with the O-2A progenitor phenotype (O4 + only) were increased in number at one week post viral inoculation (1 WPI) and were the only immunostained cells labeled at that time by a 2-h in vivo pulse of tritiated thymidine. Both GFAP+ only and GFAP+, O4+ astrocytes were also increased in the spinal cord at 1 WPI. Between two and four WPI, the infected spinal cord was characterized by the loss of (CNP+, O4+) oligodendrocytes within demyelinating lesions and the presence of O-2A progenitor cells and O4+, GFAP+ astrocytes, both of which could be labeled with thymidine. As remyelination proceeded, CNP immunostaining returned to near normal and tritiated thymidine injected previously during the demyelinating phase now appeared in CNP+ oligodendrocytes. Thus O4 positive O-2A progenitor cells proliferate early in the course of the demyelinating disease, while CNP positive oligodendrocytes do not. The timing of events suggests that the O-2A progenitors may give rise to new oligodendrocytes and to type 2 astrocytes, both of which are likely to be instrumental in the remyelination process.