Chemical traumatization of adult mouse olfactory epithelium in situ stimulates growth and differentiation of olfactory neurons in vitro.
Chemical traumatization of adult mouse olfactory epithelium in situ stimulates growth and differentiation of olfactory neurons in vitro.
复制标题
原位成年小鼠嗅觉上皮的化学创伤刺激体外嗅觉神经元的生长和分化。
DOI：
10.1016/0006-8993(95)00960-7
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发表时间：
1995
期刊：
影响因子：
2.9
通讯作者：
Roisen,FJ
中科院分区：
文献类型：
--
作者：
Sosnowski,JS;Gupta,M;Reid,KH;Roisen,FJ
本研究表明，在体外培养条件下，锌诱导的化学损伤可导致嗅觉上皮细胞的原位再生，为嗅觉神经元提供丰富的群体。因此，嗅觉上皮对化学损伤的反应能力随着有丝分裂活性的增加而增加，可以用来促进培养中神经元的生长。从正常或赋形剂处理的成年小鼠获得的组织在培养中保持时产生的嗅觉神经元比从原位硫酸锌损伤后的组织培养得到的组织少。从化学创伤后4-6天取出的组织建立的培养获得的神经元产量最高。用形态观察和细胞特异性中间丝蛋白的存在来区分这些嗅觉上皮细胞的类型。角蛋白免疫阳性，神经丝蛋白和GFAP免疫阴性的单个细胞和细胞团被鉴定为上皮样细胞。GFAP免疫阳性的扁平多角形细胞为胶质细胞。一小部分扁平细胞对本研究中使用的所有抗体均呈免疫阴性。有突起的细胞GFAP和角蛋白免疫反应为阴性。有些对200 kDa和160 kDa的神经丝蛋白呈免疫阳性反应，而对68 kDa的神经丝蛋白呈阴性反应。其中少数细胞对嗅觉标记蛋白(OMP)抗体呈阳性反应，很可能代表了培养中最成熟的嗅觉神经元。这种创伤诱导的成年小鼠嗅觉组织培养模型可以作为中枢神经系统神经元的来源，与培养的胚胎神经元进行比较。
This study demonstrates that ZnSO4induced chemical trauma results in an in situ regeneration of the olfactory epithelium which, when maintained in vitro, provides an enriched population of olfactory neurons. Therefore, the ability of the olfactory epithelium to respond to chemical trauma with increased mitotic activity can be used to increase growth of neurons in culture. Tissue obtained from normal or vehicle-treated adult mice produced few olfactory neurons, when maintained in culture, compared to cultures established from tissue following an in situ ZnSO4trauma. Maximal neuronal yields were obtained in cultures established from tissue that was removed 4–6 days following chemical trauma. The morphological appearance and the presence of cell specific intermediate filament proteins were used to classify the cell types in these olfactory epithelial cultures. Single cells and aggregates of cells which were immunopositive for keratin, but immunonegative for neurofilament protein and GFAP, were identified as epithelioid. Flattened polygonal cells immunopositive for GFAP were identified as glia. A small population of flattened cells was immunonegative for all of the antibodies used in this study. Cells that had processes were immunonegative for GFAP and keratin. Some were immunopositive for 200 kDa and 160 kDa neurofilament proteins but immunonegative for the 68 kDa neurofilament protein. A few of these cells showed positive immunoreactivity with the olfactory marker protein (OMP) antibody and most likely represented the most mature olfactory neurons in the cultures. This trauma-induced culture model using olfactory tissue from adult mice can serve as a source of CNS neurons for comparison with cultured embryonic neurons.