Endogenous arginine vasopressin–positive retinal cells in arginine vasopressin–eGFP transgenic rats identified by immunohistochemistry and reverse transcriptase–polymerase chain reaction
Endogenous arginine vasopressin–positive retinal cells in arginine vasopressin–eGFP transgenic rats identified by immunohistochemistry and reverse transcriptase–polymerase chain reaction
复制标题
通过免疫组织化学和逆转录聚合酶链反应鉴定精氨酸加压素-eGFP转基因大鼠的内源性精氨酸加压素阳性视网膜细胞
作者：
Satoru Moritoh;Kaori Sato;Y. Okada;A. Koizumi
目的精氨酸加压素（AVP）在神经组织中除了具有血管收缩作用外，还具有多种功能。以往的一些研究表明，AVP在视网膜中的存在，但AVP的来源尚未确定。本研究的目的是解决视网膜细胞是否有能力合成内源性AVP的旁分泌或自分泌的方式发挥作用的问题。方法采用AVP-eGFP转基因大鼠，用AVP抗体和GFP抗体免疫组织化学方法检测视网膜内内源性AVP阳性细胞。我们还研究了AVP mRNA和AVP受体基因的逆转录酶（RT）-PCR分离的GFP阳性细胞和整个视网膜。结果免疫组化结果显示，AVP-eGFP转基因大鼠视网膜神经节细胞层和内核层可见内源性AVP阳性细胞。分离的GFP阳性细胞的RT-PCR结果表明，这些细胞具有合成内源性AVP的能力，以及转基因AVP-eGFP。此外，通过全视网膜RT-PCR，在野生型大鼠视网膜中检测到V1 a和V1 b AVP受体，但未检测到V2受体。在分离的AVP-eGFP阳性细胞中，通过RT-PCR检测不到AVP受体。此外，AVP分泌没有检测到高钾（50 mM）溶液的刺激。结论在大鼠视网膜中发现了具有合成内源性AVP能力的细胞，并存在V1 a和V1 b型AVP受体。两者合计，这些结果表明，视网膜有一个内在的AVP合成和接收机制，可以通过V1 a和V1 b受体的旁分泌方式运作。
Purpose Recently, arginine vasopressin (AVP) has been revealed to have diverse functional roles in nervous tissues beyond that of a vasoconstrictor. Several previous studies have indicated the existence of AVP in the retina, but the source of AVP has not been determined. The objective of the present study was to address the question of whether retinal cells have the ability to synthesize endogenous AVP to act in a paracrine or autocrine manner. Methods We used AVP-eGFP transgenic rats to find endogenous AVP-positive cells in the retina by immunohistochemistry with an AVP antibody and a GFP antibody. We also examined AVP mRNA and AVP receptor genes by reverse transcriptase (RT)–PCR of dissociated GFP-positive cells and whole retinas. Results Endogenous AVP-positive cells were found in the ganglion cell layer and inner nuclear layer of the retina of AVP-eGFP transgenic rats by immunohistochemistry. As indicated by the results of RT–PCR of dissociated GFP-positive cells, these cells have the ability to synthesize endogenous AVP, as well as transgenic AVP-eGFP. In addition, the V1a and V1b AVP receptors were found in the wild-type rat retina by whole retina RT–PCR, but the V2 receptor was not detectable. In dissociated AVP-eGFP-positive cells, no AVP receptor was detected by RT–PCR. Moreover, AVP secretion was not detected by stimulation with a high potassium (50 mM) solution. Conclusions In the rat retina, we found retinal cells that have the ability to synthesize endogenous AVP, and that the retina possesses V1a and V1b AVP receptors. Taken together, these results suggest that the retina has an intrinsic AVP-synthesizing and -receiving mechanism that can operate in a paracrine manner via V1a and V1b receptors.
DOI：
10.1016/s0169-328x(02)00448-5
发表时间：
2002-10-30
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
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作者：
Foletta, VC;Brown, FD;Young, WS
通讯作者：
Young, WS
人加压素受体的分子药理学。
影响因子：
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作者：
Thibonnier,M;Conarty,DM;Preston,JA;Wilkins,PL;Berti-Mattera,LN;Mattera,R
通讯作者：
Mattera,R