Kobayashi H, Iwasaki T, Izumi F, Kurimoto S
Department of Pharmacology and Ophthalmology, University of Occupational and Environmental Health, Japan, School of Medicine, Kitakyushu.
Neuroscience. 1988 Jan;24(1):237-40. doi: 10.1016/0306-4522(88)90327-2.
In bovine retinal microvessels, alpha 1, alpha 2- and beta-adrenergic receptors were characterized by binding assay, using [3H]prazosin, [3H]para-aminoclonidine and [125I]iodocyanopindolol as radioligands, respectively. The microvessels were purified from bovine eyes by differential centrifugation through a high concentration of bovine serum albumin followed by use of a glass bead filtration technique. In the preparation, specific binding sites for [3H]para-aminoclonidine and [125I]iodocyanopindolol were observed, whereas [3H]prazosin binding was not detected. The [3H]para-aminoclonidine binding sites localized to the microvessels were characterized by high affinity and saturability (KD: 173 +/- 9 pM; Bmax: 394 +/- 11 fmol/mg protein) as well as the [125I]iodocyanopindolol binding sites (KD: 20 +/- 3 pM; Bmax: 43 +/- 4 fmol/mg protein). Furthermore, the specificity of both binding sites was pharmacologically evaluated by measuring the inhibitory effects of various adrenergic reagents on binding. The existence of alpha 2- and beta-adrenergic receptors which were characterized by high affinity, saturability and stereospecificity, leads to the hypothesis that the retinal microcirculation is under neuronal control.
在牛视网膜微血管中,分别使用[3H]哌唑嗪、[3H]对氨基可乐定和[125I]碘氰吲哚洛尔作为放射性配体,通过结合试验对α1、α2和β肾上腺素能受体进行了表征。微血管通过在高浓度牛血清白蛋白中进行差速离心,然后使用玻璃珠过滤技术从牛眼中纯化得到。在该制剂中,观察到了[3H]对氨基可乐定和[125I]碘氰吲哚洛尔的特异性结合位点,而未检测到[3H]哌唑嗪的结合。定位于微血管的[3H]对氨基可乐定结合位点具有高亲和力和饱和性(KD:173±9 pM;Bmax:394±11 fmol/mg蛋白),[125I]碘氰吲哚洛尔结合位点也是如此(KD:20±3 pM;Bmax:43±4 fmol/mg蛋白)。此外,通过测量各种肾上腺素能试剂对结合的抑制作用,对两个结合位点的特异性进行了药理学评估。具有高亲和力、饱和性和立体特异性的α2和β肾上腺素能受体的存在,引发了视网膜微循环受神经控制的假说。