Müller A R, Gerstberger R
Max-Planck-Institut für Physiologische und Klinische Forschung, Bad Nauheim, Federal Republic of Germany.
Cell Tissue Res. 1992 Apr;268(1):99-107. doi: 10.1007/BF00338058.
In the present study, we have employed the monoradioiodinated alpha 2-agonist clonidine ([125I]-CLO) to characterize duck hypothalamic alpha 2-adrenoceptors and to localize alpha 2-specific binding sites in the duck brain. To validate the alpha 2-specificity of [125I]-CLO using an enriched duck hypothalamic membrane fraction, a radioreceptor assay was established by altering the membrane protein concentration, time, temperature and ionic milieu of incubation, and in the presence or absence of protease inhibitors. Competitive displacement studies revealed the following sequence of potency to displace [125I]-CLO: yohimbine greater than (-)-epinephrine greater than clonidine greater than (-)-norepinephrine greater than phentolamine greater than (-)-phenylephrine greater than (-)-isoproterenol greater than prazosin. The non-hydrolyzable guanosine 5'-triphosphate analog guanylylimidodiphosphate markedly inhibited [125I]-CLO binding suggestive of G-protein involvement. With regard to the histological distribution, diencephalic structures, such as the habenula and the nucleus reticularis of the thalamus, were densely labeled by [125I]-CLO. In the hypothalamus, alpha 2-adrenoceptors were detected in the antidiuretic hormone-synthesizing nucleus paraventricularis, the nucleus praeopticus medialis, the nucleus anterior medialis hypothalami, the nucleus magnocellularis praeopticus, the nucleus commissurae pallii, the nucleus inferior hypothalami and the regio lateralis hypothalami. Circumventricular organs, such as the plexus choroidei, organum subfornicale, organum paraventriculare and the corpus pineale, were endowed with alpha 2-specific binding sites, as were the cell layers of the tectum opticum. In addition, telencephalic structures revealed high receptor densities.(ABSTRACT TRUNCATED AT 250 WORDS)
在本研究中,我们使用单放射性碘化α2-激动剂可乐定([125I]-CLO)来表征鸭下丘脑α2-肾上腺素能受体,并在鸭脑中定位α2特异性结合位点。为了使用富集的鸭下丘脑膜组分验证[125I]-CLO的α2特异性,通过改变膜蛋白浓度、时间、温度和孵育的离子环境,以及在有或没有蛋白酶抑制剂的情况下,建立了放射性受体测定法。竞争性置换研究揭示了以下置换[125I]-CLO的效力顺序:育亨宾大于(-)-肾上腺素大于可乐定大于(-)-去甲肾上腺素大于酚妥拉明大于(-)-苯肾上腺素大于(-)-异丙肾上腺素大于哌唑嗪。不可水解的鸟苷5'-三磷酸类似物鸟苷酰亚胺二磷酸显著抑制[125I]-CLO结合,提示G蛋白参与。关于组织学分布,间脑结构,如缰核和丘脑网状核,被[125I]-CLO密集标记。在下丘脑中,在抗利尿激素合成的室旁核、视前内侧核、下丘脑前内侧核、视前大细胞性核、缰连合核、下丘脑下部核和下丘脑外侧区检测到α2-肾上腺素能受体。室周器官,如脉络丛、穹窿下器官、室旁器官和松果体,以及视顶盖的细胞层都具有α2特异性结合位点。此外,端脑结构显示出高受体密度。(摘要截短于250字)