CELL SORTING AND ANTIBODIES TO NEUROENDOCRINE CELLS

这项研究的长期目标是增加对 细胞表面在神经内分泌发育和功能中的作用 细胞，特别是下丘脑大细胞神经分泌神经元。 的 主要目的是确定是否有质膜分子， 区分神经分泌细胞的群体，并利用任何这样的 分子来产生细胞类型特异性标记。 该问题将被 通过使用单克隆抗体（MAb）作为抗原性的探针来解决 决定因素在质膜。 因为这些分子无疑代表了 提取物中抗原分子的一小部分 神经组织，提出了几种新的技术，以提高 获得针对它们的MAb的可能性。 两大战略将是 采用;神经分泌细胞表面免疫原富集 分子，并在与融合前选择活化的淋巴细胞 骨髓瘤伴侣。 这两种策略都利用了荧光激活 细胞分选（FACS）技术。 这种MAb的成功生产将使测试几种MAb成为可能。 具体的假设。 其中包括：（1）特定细胞表面 分子介导严格的化学亲和机制发挥更重要的 竞争性的电路重排的作用， 下丘脑-神经垂体发育;（2）糖皮质激素和 其它激素或营养因子影响细胞存活和/或肽 神经分泌分化过程中的基因表达;和（3） 质膜分子浓度的改变介导了 在成熟的大细胞中发生明显的塑性变化， 神经分泌系统 本研究的一个技术目标是应用新的流式细胞仪技术， 神经内分泌细胞分析。 方法已被开发用于 纯化稀有神经元群体（固定的含神经垂体素的 神经元）用作免疫原，以尝试产生针对它们的 表面抗原 此外，使用FACS作为融合前的工具， 正在探索活化淋巴细胞的选择，以增加 产生针对稀有抗原的单克隆抗体的可能性。 这项研究的结果应该增加我们对 正常个体发育和神经分泌功能的细胞表面 神经元 此外，它们还可能揭示一些病理状态， 特发性尿崩症

The long term objective of this research is to increase understanding of the role of the cell surface in development and function of neuroendocrine cells, specifically hypothalamic magnocellular neurosecretory neurons. The primary aim is to determine if there are plasma membrane molecules which distinguish populations of neurosecretory cells and to utilize any such molecules to produce cell-type-specific markers. This question will be addressed by using monoclonal antibodies (MAbs) as probes for antigenic determinants in the plasmalemma. Because such molecules no doubt represent a very small proportion of the total antigenic molecules in extracts of neural tissue, several novel techniques are proposed to enhance the probability of obtaining MAbs against them. Two general strategies will be employed; enrichment of immunogens in neurosecretory cell surface molecules, and selection for activated lymphocytes prior to fusion with myeloma partners. Both strategies take advantage of fluorescence activated cell sorting (FACS) technology. Successful production of such MAbs would make it possible to test several specific hypotheses. Among these are: (1) that specific cell surface molecules mediating strict chemoaffinity mechanisms play a more important role than competitive rearrangement of circuitry during hypothalamic-neurohypophysial development; (2) that glucocorticoids and other hormonal or trophic factors influence cell survival and/or peptide gene expression during neurosecretory differentiation; and (3) that alterations in the concentrations of plasmalemmal molecules mediate the marked plastic changes which occur in the mature magnocellular neurosecretory system. A technical aim of this research is to apply novel FACS techniques to analysis of neuroendocrine cells. Methods have been developed for purification of rare neuronal populations (fixed neurophysin-containing neurons) for use as immunogens in attempts to produce MAbs against their surface antigens. In addition, use of the FACS as a tool for pre-fusion selection of activated lymphocytes is being explored in order to increase the probability of producing MAbs to rare antigens. Results of this research should increase our understanding of the role of the cell surface in normal ontogeny and function of neurosecretory neurons. In addition, they might shed light on such pathological states as idiopathic diabetes insipidus.