Center for Addiction Research and Department of Pharmacology and Toxicology , University of Texas Medical Branch , Galveston , Texas 77555 , United States.
ACS Chem Neurosci. 2019 Jul 17;10(7):3241-3248. doi: 10.1021/acschemneuro.8b00669. Epub 2019 Mar 18.
The 5-HT receptor (5-HTR) and 5-HTR are localized to the same neurons within the medial prefrontal cortex (mPFC), which regulates executive function, decision-making, and reward-guided learning and memory processes. The 5-HTR and 5-HTR coimmunoprecipitate in the mPFC of male Sprague-Dawley rats, while in vitro studies demonstrate the presence of a physical interaction between the 5-HTR and 5-HTR. The purpose of this study was to identify mPFC subregions in which the 5-HTR and 5-HTR physically interact ex vivo in the male Sprague-Dawley rat. We established the expression patterns of 5-HTR and 5-HTR in layers I-VI of the anterior cingulate cortex (ACC), prelimbic (PL), and infralimbic (IL) subregions using double-label fluorescence immunohistochemistry in male rats. We then employed the proximity ligation assay (PLA) to test the hypothesis that the 5-HTR and 5-HTR form a close, physical association within these mPFC subregions. Our results demonstrate subregion- and layer-specific expression of the 5-HTR and 5-HTR proteins using immunofluorescence and single recognition PLA, and a spatially close (within 40 nm) interaction between the 5-HTR and 5-HTR that occurs along a dorsal-ventral gradient in the rat mPFC.
5-羟色胺受体（5-HTR）和 5-HTR 位于内侧前额叶皮层（mPFC）的相同神经元内，调节执行功能、决策、奖励导向学习和记忆过程。5-HTR 和 5-HTR 在雄性 Sprague-Dawley 大鼠的 mPFC 中共免疫沉淀，而体外研究表明 5-HTR 和 5-HTR 之间存在物理相互作用。本研究的目的是确定雄性 Sprague-Dawley 大鼠 mPFC 亚区中 5-HTR 和 5-HTR 体外物理相互作用的位置。我们使用雄性大鼠的双标荧光免疫组织化学技术，在扣带回前皮质（ACC）、前额叶皮质（PL）和下边缘皮质（IL）的 I-VI 层中建立了 5-HTR 和 5-HTR 的表达模式。然后，我们采用邻近连接检测（PLA）来检验 5-HTR 和 5-HTR 在这些 mPFC 亚区中形成紧密的物理关联的假设。我们的结果使用免疫荧光和单识别 PLA 显示了 5-HTR 和 5-HTR 蛋白的亚区和层特异性表达，以及 5-HTR 和 5-HTR 之间在大鼠 mPFC 中沿背腹梯度发生的空间上接近（<40nm）相互作用。
