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。这个 主要目的是确定是否存在质膜分子 区分神经分泌细胞的群体并利用任何这样的 分子产生特定细胞类型的标记。这个问题将是 用单抗(MAb)作为抗原性的探针 质膜中的决定因素。因为这样的分子无疑代表着 在红豆杉提取物中占总抗原分子的很小一部分 神经组织，提出了几种新的技术来增强 获得针对它们的单抗的概率。两个一般战略将是 免疫原在神经分泌细胞表面的浓缩 分子，以及融合前活化淋巴细胞的选择 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。 这种单抗的成功生产将使其有可能测试几个 具体的假设。其中包括：(1)特定的细胞表面 调节严格的化学亲和力机制的分子发挥着更重要的作用 比电路的竞争性重排更重要的作用 下丘脑-神经垂体发育；(2)糖皮质激素和 其他激素或营养因子影响细胞存活和/或多肽 神经分泌分化过程中的基因表达； 血浆膜分子浓度的变化介导了 在成熟的巨噬细胞中发生的显著的可塑性变化 神经分泌系统。 这项研究的技术目标是将新的FACS技术应用于 神经内分泌细胞分析。已经开发出了用于 稀有神经元群的纯化(含固定神经物理素类 神经元)作为免疫原，试图产生针对其神经元的单抗 表面抗原。此外，使用FACS作为预融合的工具 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抗的可能性。 这项研究的结果应该会增加我们对 正常个体发育中的细胞表面与神经分泌功能 神经元。此外，它们还可能揭示病态，如 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.