NEUROGENESIS IN THE OLFACTORY AND VOMERONASAL SYSTEMS

嗅觉和犁鼻系统的神经发生

• 批准号: 2283993
• 负责人: David A. Holtzman
• 金额: $ 7.01万
• 依托单位: OBERLIN COLLEGE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2283993
• 关键词: alternatives to animals in research; autoradiography; biomarker; cell cycle; cell cycle proteins; cell differentiation; chemoreceptors; developmental genetics; developmental neurobiology; embryo /fetus tissue /cell culture; epidermal growth factor; gene expression; growth factor receptors; immunocytochemistry; in situ hybridization; limbic system; microtubule associated protein; neural cell adhesion molecules; neural growth associated protein; neurogenesis; neurotrophic factors; olfactions; olfactory lobe; respiratory epithelium; snakes; transcription factor

The long-term objective of this research is to understand the factors controlling neurogenesis and cell differentiation in the olfactory (OS) and vomeronasal (VNS) systems. These two major nasal chemosensory systems, found in most terrestrial vertebrates, are the only sensory systems that are known to send information directly to the telencephalon, and they do so via completely separate pathways. The olfactory (OE) and vomeronasal (VNE) epithelia of both mammals and reptiles are similar in that both contain bipolar neurons that turnover during adulthood and show target-specific regeneration following section of their respective nerves. Thus, the OE and VNE of mammals and reptiles have been used successfully in studies of postnatal neurogenesis and comparisons between regeneration and development. Snakes are a valuable research resource for investigating developmental processes in the OS and VNS. Embryonic snakes can be grown in culture, facilitating experimental manipulations. OS and VNS structures are hypertrophied through the telencephalon and mediate a wide variety of snake behaviors. Ultimately, I hope to examine the consequences of cell differentiation, the OS and VNS, on neonatal and embryonic behavior. The specific aims of this research proposal are: 1) to identify stem cells in the OS and VNS of snakes, using markers for cell cycle events, 2) to determine the fate of neuronal precursors during normal development and 3) to identify the spatial and temporal distributions of epidermal (EGF) and neuronal (NGF) growth factor receptors and their relationship to fos expression in OS and VNS structures of snakes. Using immunocytochemistry (ICC) and 3H-thymidine autoradiography (ARG), the purpose of the first aim is to establish a method for identifying reliably stem cells within OS and VNS structures. I will determine how and if the number of neuronal precursors changes during development of the receptor epithelia and telencephalic structures of the OS and VNS. By double labelling cells with cell cycle markers and 3H-thymidine ARG, correlations between cell cycle marker expression and DNA synthesis will examine the fate of cells incorporating 3H-thymidine during the course of normal development by increasing survival times after 3H-thymidine administration into embryos grown in vitro and into neonates. The state of differentiation will be assessed using ICC with antibodies and in situ hybridization (ISH) with cDNA probes to markers for neuronal development and maturation. These studies will establish the time course of neuronal differentiation in OS and VNS structures. The third aim will determine if OS and VNS structures express EGF and NGF receptors and determine if their temporal expression correlates to periods of differentiation. The expression of the immediate-early gene, fos, will be correlated to the appearance of growth factor receptors. The results of this research plan should shed light on how cell cycle control and growth factors influence the differentiation and maturation of neuronal precursors.

这项研究的长期目标是了解影响因素 控制嗅觉 (OS) 中的神经发生和细胞分化 和犁鼻 (VNS) 系统。 这两个主要的鼻化学感应 在大多数陆生脊椎动物中发现的系统是唯一的感觉系统 已知直接向端脑发送信息的系统， 他们通过完全独立的途径来做到这一点。 嗅觉 (OE) 和 哺乳动物和爬行动物的犁鼻（VNE）上皮细胞在以下方面相似： 两者都含有双极神经元，这些神经元在成年期间会翻转并显示 特定目标的再生遵循各自的部分 神经。 因此，哺乳动物和爬行动物的OE和VNE已被使用 成功地进行了产后神经发生的研究和比较 再生和发展。 蛇是宝贵的研究资源 用于研究操作系统和 VNS 的开发过程。 胚胎 蛇可以在培养物中生长，有利于实验操作。 OS 和 VNS 结构通过端脑肥大， 介导蛇的多种行为。 最后，我希望能够检查一下 细胞分化、OS 和 VNS 对新生儿和新生儿的影响 胚胎行为。 本研究计划的具体目标是：1）确定干 蛇的 OS 和 VNS 中的细胞，使用细胞周期事件的标记， 2）确定神经元前体在正常发育过程中的命运 3）确定表皮的空间和时间分布 (EGF) 和神经元 (NGF) 生长因子受体及其关系 fos 在蛇的 OS 和 VNS 结构中的表达。 使用 免疫细胞化学 (ICC) 和 3H-胸苷放射自显影 (ARG) 第一个目标的目的是建立一种识别方法 OS 和 VNS 结构中可靠的干细胞。 我将决定如何 如果神经元前体的数量在发育过程中发生变化 OS 和 VNS 的受体上皮和端脑结构。 通过使用细胞周期标记物和 3H-胸苷 ARG 对细胞进行双标记， 细胞周期标记表达与 DNA 合成之间的相关性 检查过程中掺入 3H-胸苷的细胞的命运 通过增加 3H-胸苷后的存活时间来促进正常发育 施用到体外生长的胚胎和新生儿中。 国家 将使用带有抗体的 ICC 和原位评估分化的情况 与神经元发育标记物的 cDNA 探针杂交 (ISH) 和成熟。 这些研究将确定神经元的时间进程 OS 和 VNS 结构的差异。 第三个目标将决定 OS和VNS结构是否表达EGF和NGF受体并确定是否 它们的时间表达与分化时期相关。 这 立即早期基因 fos 的表达将与 生长因子受体的出现。 本研究计划的成果 应该阐明细胞周期控制和生长因子如何影响 神经元前体的分化和成熟。