权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Expression of CRF and iNOS mRNA in the hypothalamus and the mode of response to stress.

下丘脑CRF和iNOS mRNA的表达及应激反应模式。

基本信息

批准号：
11671657
负责人：
AKIRA Shigeo
金额：
$ 1.41万
依托单位：
Nippon Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671657/
关键词：
CRF iNOS rat fetus LPS in situ hybridization 胎仔 NOS ラット mRNA CRFレセプター

项目摘要

We investigated the expression of CRF and iNOS mRNA, and the mode of response to lipopolysaccharide (LPS) loading in rat embryo models for infection. Embryos were delivered by cesarean section at day 20 of pregnancy and immediately placed in a chamber with the temperature controlled at 37℃ and the humidity at 100%. Either LPS (400μg/100g of the body weight) or physiological saline was administered to embryos, and the embryos were maintained for 3 hours. Embryos were decapitated after cesarean section and after 3 hours of maintenance. Fetal brain was collected, fixed, and prepared by coronal section. We produced a ^<35>S-UTP-labelled antisense RNA probe for CRF and iNOS, and performed in situ hybridization. The expression of CRF and iNOS was analyzed by optical density. Moreover, blood corticosterone was assayed by radioimmunoassay. The expression of iNOSmRNA was not seen in the control group, but the expression was identified in the subfornical organs and the choroids plexus, although no expression was seen in the paraventricular nucleus in the LPS group. Significantly increased blood corticosterone was observed in the LPS group. In conclusion, in 20-day-old embryos, the CRF-ACTH-corticosterone pathway has been activated in response to the stress of LPS loading. However, the absence of expression of iNOSmRNA on the paraventricular nucleus suggests that NO is not the major stress-transmission mediator in stimulating CRF release in response to LPS loading.

我们研究了 CRF 和 iNOS mRNA 的表达，以及感染大鼠胚胎模型中对脂多糖 (LPS) 负荷的反应模式。妊娠第20天剖腹产胚胎，立即放入温度控制在37℃、湿度100%的培养箱中。将LPS(400μg/100g体重)或生理盐水给予胚胎，并将胚胎维持3小时。剖腹产后和维持3小时后将胚胎斩首。收集、固定并通过冠状切片制备胎儿大脑。我们制备了CRF和iNOS的35S-UTP标记的反义RNA探针，并进行了原位杂交。通过光密度分析CRF和iNOS的表达。此外，通过放射免疫测定法测定血液皮质酮。对照组未见iNOSmRNA表达，但LPS组室旁核未见iNOSmRNA表达，但穹窿下器官和脉络丛中有表达。 LPS 组观察到血皮质酮显着升高。总之，在 20 天大的胚胎中，CRF-ACTH-皮质酮途径已被激活以响应 LPS 负荷的压力。然而，室旁核上 iNOSmRNA 的缺失表明，NO 并不是响应 LPS 负荷而刺激 CRF 释放的主要应激传递介质。