NEUROGENESIS IN THE OLFACTORY AND VOMERONASAL SYSTEMS

这项研究的长期目标是了解这些因素 控制嗅觉(OS)的神经发生和细胞分化 和犁鼻(VNS)系统。这两种主要的鼻腔化学感觉 在大多数陆地脊椎动物中发现的系统是唯一的感官 已知的将信息直接发送到端脑的系统， 它们通过完全不同的路径完成这一过程。嗅觉(OE)和 哺乳动物和爬行动物的犁鼻上皮在以下方面相似 两者都含有双极神经元，在成年期更替并表现出 特定于目标的再生遵循其各自的 太紧张了。因此，哺乳动物和爬行动物的OE和VNE被使用 在出生后神经发生的研究中取得成功，并与 再生发展。蛇是一种宝贵的研究资源。 用于研究操作系统和VNS中的发展过程。胚胎 蛇可以在培养中生长，便于进行实验性的操作。 OS和VNS结构通过端脑和 调节各种各样的蛇的行为。最终，我希望研究一下 细胞分化、OS和VNS对新生儿和 胚胎行为。 这项研究计划的具体目的是：1)识别茎 蛇的OS和VNS中的细胞，使用细胞周期事件的标记， 2)确定神经前体细胞在正常发育过程中的命运 3)识别表皮的空间和时间分布 表皮生长因子和神经生长因子受体及其相互关系 FOS在蛇的OS和VNS结构中的表达。vbl.使用 免疫细胞化学(ICC)和~3H-胸腺嘧啶核苷放射自显影(ARG)； 第一个目标的目的是建立一种识别 OS和VNS结构内可靠的干细胞。我会决定如何 如果神经元前体细胞的数量在发育过程中发生变化 OS和VNS的受体上皮细胞和端脑结构。 通过用细胞周期标记物和~3H-胸腺嘧啶核苷ARG双重标记细胞， 细胞周期标志物表达与DNA合成的相关性 检测在此过程中掺入~3H-胸腺嘧啶核苷的细胞的命运 通过延长~3H-胸腺嘧啶核苷治疗后的存活时间来恢复正常发育 对体外培养的胚胎和新生儿进行注射。国家 分化程度的评估将使用ICC与抗体和原位 神经元发育标志物的基因探针杂交(ISH) 和成熟。这些研究将建立神经元的时间进程。 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 它们在时间上的表达与分化时期相关。这个 即刻早期基因fos的表达将与 生长因子受体的出现。这项研究计划的结果 应该阐明细胞周期控制和生长因素是如何影响的 神经前体细胞的分化和成熟。

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.