Characterization of antisera raised against cultured rat sympathetic neurons.
Characterization of antisera raised against cultured rat sympathetic neurons.
复制标题
针对培养的大鼠交感神经元产生的抗血清的表征。
DOI：
10.1016/0306-4522(80)90140-2
复制
发表时间：
1980
期刊：
影响因子：
3.3
通讯作者：
Greene,LA
中科院分区：
文献类型：
--
作者：
Lee,VM;Shelanski,ML;Greene,LA
制备了抗新生大鼠交感神经细胞单层培养的抗血清，并用~(51)Cr释放微量补体结合试验、细胞毒性试验和间接免疫荧光试验对其进行了表征。该抗血清经大鼠肝、肾、脾、胸腺吸收去除对常见大鼠抗原的活性后，对培养的交感神经元、脑和肾上腺仍有活性。间接免疫荧光显示，识别的抗原广泛分布于脑内，而在肾上腺中仅限于髓细胞。与大脑的吸收完全消除了抗血清对交感神经元的活性；相反，在与肾上腺重复吸收后保留了部分活性。抗血清对大鼠嗜铬细胞瘤细胞株PC12的识别作用也进行了研究。在正常生长条件下，PC12细胞类似于正常的肾上腺嗜铬细胞，而经神经生长因子处理后，它们获得交感神经元的分化特性。未处理和处理的细胞均可被抗血清识别，活细胞上的间接组织荧光表明，这些细胞(以及交感神经元上的)至少有部分抗原暴露在细胞表面。然而，在神经生长因子处理和未处理的PC12细胞的识别方面存在差异。例如，前者在细胞毒性试验中被识别，而后者不被识别。这些发现表明，经神经生长因子处理的PC12细胞和交感神经元表现出抗原相似性，我们的抗血清可用于鉴定、分离、表征和定位神经特异性表面抗原。
Antisera were prepared against monolayer cultures of newborn rat sympathetic neurons and characterized by51Cr release microcomplement fixation, cytotoxicity and indirect immunofluorescence assays. After absorption with rat liver, kidney, spleen and thymus to remove activity against common rat antigens, the antisera retained activity against cultured sympathetic neurons, brain and adrenals. Indirect immunofluorescence showed that the antigens recognized were widely distributed in the brain, while in the adrenal gland they were limited to the medullary cells. Absorption with the brain completely removed the activity of the antisera against sympathetic neurons; in contrast, partial activity was retained after repetitive absorption with adrenals. The antisera were also characterized with respect to recognition of the PC12 clonal line of rat pheochromocytoma cells. Under normal growth conditions, PC12 cells resemble their normal counterparts, adrenal chromaffin cells, while after treatment with nerve growth factor, they acquire the differentiated properties of sympathetic neurons. Both untreated and treated cells were recognized by the antisera and indirect histofluorescence on living cells indicated that at least some of the antigens on these cells (as well as on sympathetic neurons) are exposed on the cell surface. However, differences were noted in recognition of nerve growth factor treated and untreated PC12 cells. For example, the former, but not the latter were recognized in cytotoxicity assays. Also, absorption of the antisera with treated cells removed all activity against rat sympathetic neurons, whereas repetitive absorption with untreated PC12 cells removed only part of the activity.Such findings indicate that PC12 cells treated with nerve growth factor and sympathetic neurons exhibit antigenic similarities and that our antisera can be used to identify, isolate, characterize and localize neural-specific surface antigens.