Mastrodimou N, Vasilaki A, Papadioti A, Low M J, Hoyer D, Thermos K
University of Crete, Faculty of Medicine, Department of Basic Sciences, Laboratory of Pharmacology, Heraklion, Crete 71110, Greece.
Neuropeptides. 2006 Oct;40(5):365-73. doi: 10.1016/j.npep.2006.07.002. Epub 2006 Sep 28.
The present study investigated the localization and density of somatostatin (SRIF) receptor subtypes (sst(1-5)) and SRIF-nitric oxide (NO()) interactions in the retina of wildtype [WT, (+/+)] and somatostatin deficient mice [SRIF (-/-)]. Immunohistochemistry and radioligand binding studies with subsequent autoradiography were performed. Monoclonal antibodies [SRIF, protein kinase C (rod bipolar cells marker), microtubule associated protein 1A (ganglion cell marker)] and polyclonal antibodies (anti-sst(1), sst(2A), sst(4) receptor) were applied to 10-14 microm sections of retinas fixed in paraformaldehyde. NADPH-diaphorase reactivity was assessed histochemically. [(125)I]LTT SRIF-28 alone or in the presence of MK678 (sst(2) agonist) and [(125)I]Tyr(3)-octreotide were employed to quantify sst(1-5), sst(1/4)and sst(2/5) receptor densities, respectively. sst(1), sst(2A), and sst(4) receptor immunoreactivities were observed in processes of the inner plexiform layer (IPL), rod bipolar, and in ganglion cells and processes, respectively, in WT and SRIF (-/-) mice. Specific [(125)I]LTT SRIF-28 and [(125)I]Tyr(3)-octreotide binding was increased significantly in SRIF (-/-) mice. NADPH-diaphorase staining was localized in photoreceptors and amacrine cells, but not rod bipolar and ganglion cells. Also, NADPH-diaphorase staining was not colocalized with sst(1), sst(2A) or sst(4) receptor immunoreactivity. These results demonstrate an upregulation of SRIF receptors in mice lacking SRIF, but no evident SRIF-NO(*) interaction was observed in the mouse retina.
本研究调查了野生型[WT, (+/+)]和生长抑素缺陷型小鼠[SRIF (-/-)]视网膜中生长抑素(SRIF)受体亚型(sst(1 - 5))的定位和密度以及SRIF与一氧化氮(NO)的相互作用。进行了免疫组织化学和放射性配体结合研究及随后的放射自显影。将单克隆抗体[SRIF、蛋白激酶C(视杆双极细胞标志物)、微管相关蛋白1A(神经节细胞标志物)]和多克隆抗体(抗sst(1)、sst(2A)、sst(4)受体)应用于固定在多聚甲醛中的10 - 14微米厚的视网膜切片。通过组织化学方法评估NADPH - 黄递酶反应性。单独使用[(125)I]LTT SRIF - 28或在MK678(sst(2)激动剂)存在的情况下使用[(125)I]LTT SRIF - 28以及[(125)I]Tyr(3)-奥曲肽分别用于定量sst(1 - 5)、sst(1/4)和sst(2/5)受体密度。在WT和SRIF (-/-)小鼠中,分别在内网状层(IPL)的突起、视杆双极细胞以及神经节细胞及其突起中观察到sst(1)、sst(2A)和sst(4)受体免疫反应性。在SRIF (-/-)小鼠中,特异性[(125)I]LTT SRIF - 28和[(125)I]Tyr(3)-奥曲肽结合显著增加。NADPH - 黄递酶染色定位于光感受器和无长突细胞,但不在视杆双极细胞和神经节细胞中。此外,NADPH - 黄递酶染色与sst(1)﹑sst(2A)或sst(4)受体免疫反应性不共定位。这些结果表明,在缺乏SRIF的小鼠中SRIF受体上调,但在小鼠视网膜中未观察到明显的SRIF - NO相互作用。