Establishment of in vitro model for the vomeronasal system and analysis of the synapse formation between the vomeronasal and accessory olfactory neurons

犁鼻系统体外模型的建立及犁鼻与副嗅神经元突触形成的分析

基本信息

批准号：
16500201
负责人：
MURAMOTO Kazuyo
金额：
$ 2.05万
依托单位：
Kochi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16500201/
关键词：
co-culture vomeronasal organ calcium imaging synapse formation 鋤鼻系 Calcium Imaging

项目摘要

Pheromones are detected by unique receptors in the vomeronasal organ (VNO), and then affect endocrinal status and behavior in various kinds of animals. Although it is well known that the VNO mediates sexually and developmentally different behavioral and neuroendocrine responses, the cellular mechanisms regulating these functions are not, much understood. The differences in pheromonal effects might be in part due to differential expression of pheromone receptors. In this project, I established the co-culture system of vomeronasal receptor neurons (VRN) and accessory olfactory bulb (AOB) neurons to assess roles of each neuronal species in the vomeronasal system on the pheromonal signal processing. Firstly, I demonstrated the synapse formation between cultured VRNs and AOB neurons using a calcium imaging technique combined with electrical stimulation and analyzing pharmacologically. Secondly, it was showed that the maturation of VRNs was induced by co-culture with dissociated AOB neurons. … More To characterize the receptor expression, I examined two representatives (VR1 and VR4) from different subfamilies of the V2R family of pheromone receptors in cultured VRNs. A western blotting analysis showed the expression of VR1 and VR4 in VRNs was induced and increased with days in co-culture, an effect which was not observed in the VRN-alone culture. Moreover, we applied charged compounds in mouse urine iontophoretically to the VRNs using a microelectrode and analyzed VRN response by a Ca^<2+> imaging method with or without cultured AOB cells. When urine compounds were ejected to the VRNs co-cultured with AOB cells with a current of -2μA, subpopulation of VRNs clearly showed long-lasting Ca^<2+> increases. Injections of a current below -5μA alone had no effect to the VRNs. Such Ca^<2+> increases were not observed without AOB cells. These results indicate that VRNs result in expressing pheromone receptors by interacting with AOB neurons, and then acquiring responsiveness to compounds in urine. Less

信息素是由选民器官（VNO）中的独特受体检测到的，然后影响各种动物的内分泌状态和行为。尽管众所周知，VNO介导了性和开发不同的行为和神经内分泌反应，但控制这些功能的细胞机制并没有太多理解。信息素效应的差异可能部分是由于信息素受体的差异表达。该项目，我建立了选民鼻受体神经元（VRN）和附属嗅球（AOB）神经元的共同培养系统，以评估每个神经元物种在选民鼻系统中的作用。首先，我使用钙成像技术与电刺激结合并进行药物分析，证明了培养的VRN和AOB神经元之间的突触形成。其次，结果表明，VRN的成熟是通过与解离的AOB神经元共培养诱导的。 …更多地表征受体表达，我检查了来自培养的VRN中V2R家族的不同亚家族的两名代表（VR1和VR4）。蛋白质印迹分析表明，VR1和VR4在VRN中的表达被诱导，并随着共培养的天数而增加，这在VRN-Alone培养物中未观察到这种作用。此外，我们使用微电极将带电的化合物在小鼠尿液离子to虫中施加到VRN，并通过具有或没有培养的AOB细胞的Ca^<2+>成像方法分析VRN响应。当将尿液化合物与AOB细胞共培养的VRNs弹出时，VRN的亚群清楚地显示了持久的Ca^<2+>的亚群。仅以低于-5μA的电流注射对VRN没有影响。没有AOB细胞，就不会观察到这种Ca^<2+>。这些结果表明，VRN通过与AOB神经元相互作用，从而表达信息素受体，然后在尿液中获得对化合物的反应性。较少的