Detection of the 5-HT1A receptor and 5-HT1A receptor mRNA in the rat bowel and pancreas: comparison with 5-HT1P receptors.
Detection of the 5-HT1A receptor and 5-HT1A receptor mRNA in the rat bowel and pancreas: comparison with 5-HT1P receptors.
复制标题
大鼠肠和胰腺中5-HT1A受体和5-HT1A受体mRNA的检测：与5-HT1P受体的比较。
DOI：
10.1002/cne.903270206
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Gershon,MD
中科院分区：
文献类型：
--
作者：
Kirchgessner,AL;Liu,MT;Howard,MJ;Gershon,MD
我们测试了大鼠肠道和胰腺含有 5-HT1A 受体的假设。3H-8-羟基-2-（二正丙氨基）四氢化萘 (3H-8-OH-DPAT) 被用作放射性配体。通过快速过滤研究 3H-8-OH-DPAT 与肌间神经丛和胰腺来源的膜的结合。或者，采用放射自显影来定位未固定肠或胰腺冰冻切片中的 3H-8-OH-DPAT 结合位点。使用过量的 5-HT (10 μM) 来定义非特异性结合。发现 3H-8-OH-DPAT 与肠膜（Kd= 2.8 ± 1.1 nM；Bmax=83.8 ± 4.3 fmol/mg 蛋白质）和胰膜（Kd= 6.6 ± 1.3 nM；Bmax= 44 ± 2.2 fmol/mg 蛋白质）具有饱和、高亲和力结合。 8-OH-DPAT、NAN-190 和螺哌隆抑制 3H-8-OH-DPAT 与肠膜和胰膜的结合。相比之下，3H-8-OH-DPAT 与肠膜和胰膜的结合不受 5-羧酰胺色胺或各种已知与 5-HT 受体其他亚型结合的化合物的抑制。地高辛标记的寡核苷酸被发现可以检测肌间神经节和粘膜下神经节的一部分神经元中编码 5-HT1A 受体的 mRNA。相比之下，在胰腺中未发现 5-HT1AmRNA。放射自显影显示，在胃中发现了最高密度的 3H-8-OH-DPAT 结合位点。这些位点在邻近胃腺的固有层和肌间神经节中尤其多。胰腺5-HT1A受体位于神经、淋巴组织（尤其是淋巴结被膜）和分散在胰腺实质中的细胞上。大鼠肠和胰腺中3H-8-OH-DPAT结合位点的浓度低于3H-5-HT结合位点的浓度；然而，3H-8-OH-DPAT 结合位点的分布与 3H-5-HT 结合位点的分布相似。结论是，大鼠肠道及其在胰腺中的延伸部分含有 5-HT1A 受体。许多（如果不是全部）表达 5-HT1A 受体的神经细胞和过程也表达 5-HT1 受体。这些受体在肠胰神经支配的生理学中的功能仍有待确定。 © 1993 Wiley-Liss, Inc.
We tested the hypothesis that the rat bowel and pancreas contain 5‐HT1Areceptors.3H‐8‐hydroxy‐2‐(di‐n‐propylamino) tetralin (3H‐8‐OH‐DPAT) was used as a radioligand. Binding of3H‐8‐OH‐DPAT to membranes derived from the myenteric plexus and the pancreas was investigated by rapid filtration. Alternatively, radioautography was employed to locate3H‐8‐OH‐DPAT binding sites in frozen sections of unfixed bowel or pancreas. An excess of 5‐HT (10 μM) was used to define nonspecific binding. Saturable, high affinity binding of3H‐8‐OH‐DPAT to enteric (Kd= 2.8 ± 1.1 nM; Bmax=83.8 ± 4.3 fmol/mgproteim) and pancreatic (Kd= 6.6 ± 1.3 nM; Bmax= 44 ± 2.2 fmol/mg protein) membranes was found. The binding of3H‐8‐OH‐DPAT to enteric and pancreatic membranes was inhibited by 8‐OH‐DPAT, NAN‐190, and spiperone. In contrast, the binding of3H‐8‐OH‐DPAT to enteric and pancreatic membranes was not inhibited by 5‐carboxyamidotryptamine, or by avariety of compounds known to bind to other subtypes of 5‐HT receptor. Digoxigenin‐labeled oligonucleotides were found to detect mRNA encoding the 5‐HT1Areceptor in a subset of neurons in myenteric and submucosal ganglia. In contrast, 5‐HT1AmRNA was not found in the pancreas. Radioautography revealed that the highest density of3H‐8‐OH‐DPAT binding sites was found in the stomach. These sites were especially numerous in the lamina propria adjacent to gastric glands, and in myenteric ganglia. Pancreatic 5‐HT1Areceptors were located on nerves, lymphoid tissue (especially the capsule of nodes), and on cells scattered in the pancreatic parenchyma. The concentration of3H‐8‐OH‐DPAT binding sites in the rat bowel and pancreas was less than that of3H‐5‐HT binding sites; however, the distribution of3H‐8‐OH‐DPAT binding sites was similar to that of sites that bind3H‐5‐HT. It is concluded that the rat gut and its extension in the pancreas contains 5‐HT1Areceptors. Many, if not all, of the nerve cells and processes that express 5‐HT1Areceptors express 5‐HT1preceptors as well. The function of these receptors in the physiology of the entero‐pancreatic innervation remains to be determined. © 1993 Wiley‐Liss, Inc.