REGULATION OF DEVELOPMENT OF PEPTIDE NEURONS IN GUT

肽能神经元在肠道中大量存在。 证据 我们和其他人最近的实验表明， 尤其是肠道，可以影响 肽表型 我们打算研究肠道中的元素 这些环境对于影响肽表达至关重要。 利用体内和体外培养技术， 鹌鹑睫状神经节-鸡神经肠组合中的神经肽。 原位睫状神经节神经元仅含有乙酰胆碱， 神经肽生长抑素 我们将用体内培养的方法研究 神经肠促进睫状体神经肽表达的能力 神经节-肠组织组合生长在绒毛尿囊膜上， 宿主鸡胚。 这一方法将允许对 肠道年龄、肠道区域和肠道神经支配对产生 神经肽在三维组织完整性和 胚胎的荷尔蒙环境得以保存。 使用体外方法，我们 将分析生长在视网膜中的睫状神经节中神经肽的表达， 不同年龄肠道的培养基。 这种方法将使我们能够 确定可溶性因子或细胞与细胞的接触是否对 肽表达。 在最后一组实验中， 和组织提取物来影响中脑神经元中的肽表达， 将研究脊文化。 一部分中脑神经 嵴是睫状神经节的前体。 肽包括 血管活性肠肽、生长抑素、脑啡肽和P物质 将通过免疫细胞化学定位并通过放射免疫测定法测量。 因为神经肽和胆碱能之间可能存在联系 表达，我们将研究神经肽和 ChAT，一种胆碱能神经元特异性的酶标记物。 与此 结合技术，我们将阐明肠道中的元素 影响神经肽表达的环境。 的信息 这里获得的将是有用的，在理解潜在的缺陷， 导致诸如先天性巨结肠的临床病症， 假性梗阻

Peptidergic neurons are found in great numbers in the gut. Evidence from our recent experiments and those of others indicates that the environment in general and the gut in particular can influence the expression of peptide phenotypes. We propose to investigate the elements in the gut environment that are critical for influencing peptide expression. Using in vivo and in vitro culture techniques, we will study the production of neuropeptides in quail ciliary ganglion-chick aneural gut combinations. Ciliary ganglion neurons in situ contain only acetylcholine and the neuropeptide somatostatin. We will study with in vivo culture methods the ability of aneural gut to promote expression of neuropeptides in ciliary ganglion-gut tissue combinations grown on the chorioallantoic membrane of host chick embryos. This approach will permit an evaluation of the variables of gut age, gut region, and gut innervation on the production of neuropeptides in a setting where the three-dimensional tissue integrity and the hormonal milieu of the embryo are preserved. Using in vitro methods we will analyze the expression of neuropeptides in ciliary ganglia grown in culture media with gut of different ages. This approach will permit us to determine if a soluble factor or cell-to-cell contact is important to peptide expression. In a last set of experiments the ability of the gut and tissue extracts to influence peptide expression in mesencephalic neural crest cultures will be studied. A portion of the mesencephalic neural crest is a precursor for the ciliary ganglion. Peptides including vasoactive intestinal peptide, somatostatin, enkephalin and substance P will be localized by immunocytochemistry and measured by radioimmunoassay. Because there may be a link between neuropeptide and cholinergic expression, we will study the possible co-localization of neuropeptides and ChAT, an enzyme marker specific for cholinergic neurons. With this combination of techniques, we will elucidate the elements in the gut environment that influence neuropeptide expression. The information obtained here will be useful in understanding the underlying defects that lead to clinical conditions such as Hirschsprung's disease and pseudo-obstruction.