Development of quantitative autoradiographic method for assessing fuctional imaging in living brain slices.

开发用于评估活体脑切片功能成像的定量放射自显影方法。

基本信息

批准号：
09557189
负责人：
SAJI Hideo
金额：
$ 5.76万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

Bio-Imaging Analyzer System with imaging plate was recently developed and possesses with ultra high sensitivity, high resolution and a wide dynamic range compared with X-ray films. Taking advantages of the Bio-Imaging Analyzer System, we examined the possibility of applying this system for functional imaging in living brain slices. Whole brain slices of 300 mum thickness were incubated with radiolabeled compounds. The slices were then rinsed and placed on the bottom of a Plexiglas chamber filled with Krebs.Ringer solution. The bottom of the chamber consisted of a thin polypropylene film to allow good penetration of radiation from the brain slices. The chamber was placed on a imaging plate. After exposure, the imaging plate was scanned by the Bio-Imaging Analyzer System and radioactivity images of brain slices were obtained. By using this system, we evaluated the glucose metabolism changes under hypoxic condition and the super high affinity binding sites in rat hippocampus for [3-H]Ro 1 … More 5-4513, a partial inverse agonist of central benzodiazepine receptors. Under hypoxic condition, increase of radioactivity of [14-C]glucose was observed in all brain regions compared with the control group. Differences of increase rate of rdioactivity uptake between brain regions were also observed. These results suggested that utilization rate of glucose increase due to acceleration of anaerobic metabolism and that energy demands vary depending on the brain regions under hypoxic condition compared with control group. On the other hand, in vitro binding of [3-H]Ro15-4513, the supar high affinity binding sites in the hippocampus were observed when the [3-H]Ro15-4513 concentration was below 0.5 nM.In vivo, the supar high affinity binding sites were only found when the injected dose of Ro15-4513 was below 3.6 mug/kg and almost disappeared when the dose was increased to 10 mug/kg. These results both in vitro and in vivo indicate that there is a significant discrepancy between actual free ligand concentration in vivo and in vitro, and that concntration in intact brain may be much lower than previously thought. Less

最近开发出带有成像板的生物成像分析系统，与X射线胶片相比，具有超高灵敏度、高分辨率和宽动态范围。利用生物成像分析系统的优点，我们研究了应用该系统的可能性。将 300 微米厚的全脑切片与放射性标记化合物一起孵育，然后将切片放置在充满 Krebs.Ringer 溶液的有机玻璃室的底部。室的底部由聚丙烯薄膜组成，以允许来自脑切片的辐射良好穿透。曝光后，通过生物成像分析仪系统扫描成像板并获取脑的放射性图像。通过使用该系统，我们评估了缺氧条件下葡萄糖代谢的变化以及大鼠海马中[3-H]Ro 1 … More 5-4513（部分逆）的超高亲和力结合位点。在缺氧条件下，与对照组相比，所有脑区的[14-C]葡萄糖放射性增加，这些结果表明，脑区之间的放射性摄取增加率存在差异。与对照组相比，由于无氧代谢的加速，葡萄糖的利用率增加，并且能量需求根据缺氧条件下的大脑区域而变化。 [3-H]Ro15-4513，当[3-H]Ro15-4513浓度低于0.5 nM时，在海马中观察到超高亲和力结合位点。在体内，只有当Ro15-4513的注射剂量低于3.6μg/kg，并且当剂量增加至10μg/kg时几乎消失。这些体外和体内结果均表明存在。体内和体外的实际游离配体浓度之间存在显着差异，并且完整大脑中的浓度可能比以前认为的要低得多。