DEVELOPMENTAL REGULATION OF NEUROPEPTIDE EXPRESSION

这份申请书是为了继续我们的研究而设计的 描述了调节基因表达的发育机制 神经肽。在Manduca(蛾)胚胎中，我们描述了 PheMetArgPhe-酰胺相关神经肽的胚胎表达 (FMRFamide)在中枢、外周和下丘脑的特定神经元 肠道神经系统。我们还分离和表征了 两种昆虫(果蝇和曼杜亚)的编码基因 FMRFamide样神经肽。进一步刻画了 Manduca基因将提供信息和探针来补充 胚胎肽能神经元在体内和体内的细胞分析 胚胎培养。 在授权期内，我们会继续发展模范系统 重点研究多肽能表型的分化。我们会 考虑遗传和表观遗传机制对 监管是以下三个基本方面的基础 差异化。第一个方面与定时机制有关 这决定了在形态发生过程中，最初的神经肽 表达式(特定于单元的属性)首先在 辨认出神经元。第二个研究的机制是 一个已鉴定神经元的暂时性神经肽表达 在胚胎发育过程中。第三个考虑延迟和位置-- 一组移行神经元中特异性神经肽的表达 在发育中的肠道中似乎不是唯一识别的。 对于每个方面，我们都将从两个方面来研究差异 神经肽含量和神经肽基因表达。按顺序 把这些观察放在更大的细胞背景下 特异神经元的分化、超微结构研究 针对神经肽前体和成熟蛋白的特异性抗体 多肽将在这些阶段的每个阶段执行 差异化。 这些研究将为以后的研究提供基础。 调节这三个方面的分子机制 差异化。此外，它们还代表先行词 迈向未来考虑细胞的实验步骤 决定，也就是，细胞和分子机制 神经肽的细胞特异性模式的产生是基础 基因表达。

This application is for a continuation of our studies designed to described development mechanisms that regulate the expression of neuropeptides. In Manduca (moth) embryos, we have described the embryonic expression of neuropeptides related to PheMetArgPhe-amide (FMRFamide) in specific neurons of the central, peripheral and enteric nervous systems. We have also isolated and characterized genes in two insects species (Drosophila and Manduca) that encode the FMRFamide-like neuropeptides. Further characterization of the Manduca gene will provide information and probes to complement the cellular analysis of embryonic peptidergic neurons in vivo and in embryo culture. During the grant period, we will continue to develop model systems to focus on the differentiation of peptidergic phenotypes. We will consider contributions of genetic and epi-genetic mechanisms to the regulation that underlies three fundamental aspects of differentiation. The first aspect pertains to the timing mechanism that dictates when, during morphogenesis, initial neuropeptide expression (a cell-specific property) is first observed in identified neurons. The second investigates the mechanisms that underlie transient neuropeptide expression in an identified neuron during embryogenesis. The third considers delayed and position- specific neuropeptide expression among a group of migrating neurons in the developing gut that do not appear to be uniquely identified. For each aspect, we will study differentiation in terms of both neuropeptide content and neuropeptide gene expression. In order to place these observation within the larger context of cellular differentiation, ultrastructural studies of specific neurons using specific antibodies to the neuropeptide precursor and to the mature peptides will be performed during each of these phases of differentiation. These studies will provide a basis for the later investigation of the molecular mechanisms that mediate these three regulated aspects of differentiation. Further, they represent antecedent experimental steps to the future consideration of cellular determination, that is, the cellular and molecular mechanisms that underlie the generation of cell-specific patterns of neuropeptide gene expression.