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因素 控制嗅觉（OS）中的神经发生和细胞分化 和Vomeronasal（VNS）系统。 这两个主要的鼻化学感应 在大多数陆地脊椎动物中发现的系统是唯一的感觉 已知可以将信息直接发送到端脑的系统， 他们通过完全独立的途径进行。 嗅觉（OE）和 两种哺乳动物和爬行动物的呕吐（VNE）上皮相似 两者都包含在成年期间营业额并显示的双极神经元 目标特异性再生之后的部分 神经。 因此，已经使用了哺乳动物和爬行动物的OE和VNE 在产后神经发生和比较的研究中成功 再生与发展。 蛇是宝贵的研究资源 用于研究OS和VNS中的发展过程。 胚胎 蛇可以在文化中种植，从而促进实验性操纵。 OS和VNS结构通过端脑肥大，并且 调解各种各样的蛇行为。 最终，我希望检查 细胞分化，OS和VN的后果，新生儿和 胚胎行为。 该研究建议的具体目的是：1）识别STEM 蛇的OS和VN中的细胞，使用标记用于细胞周期事件的标记， 2）确定正常发育过程中神经元前体的命运 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 在蛇的OS和VNS结构中表达FOS。 使用 免疫细胞化学（ICC）和3H-胸腺定自显影（ARG）， 第一个目的的目的是建立一种识别的方法 可靠的OS和VNS结构中的干细胞。 我将确定如何 以及在开发过程中神经元前体的数量变化 OS和VNS的受体上皮和脑脑结构。 通过用细胞周期标记和3H-胸腺胺ARG的双重标记细胞， 细胞周期标记表达与DNA合成之间的相关性将会 检查过程中掺入3H-胸苷的细胞的命运 通过增加3H-胸腺胺后的生存时间来正常发育 在体外生长并进入新生儿。 国家 将使用具有抗体和原位的ICC评估差异化 杂交（ISH）与cDNA探针与标志物的神经元发育 和成熟。 这些研究将确定神经元的时间过程 OS和VNS结构的分化。 第三个目标将决定 如果OS和VNS结构表达EGF和NGF受体，并确定是否是否 它们的时间表达与分化周期相关。 这 直接至前基因FOS的表达将与 生长因子受体的出现。 该研究计划的结果 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。