Distribution and characterisation of Glucagon-like peptide-1 receptor expressing cells in the mouse brain.
Distribution and characterisation of Glucagon-like peptide-1 receptor expressing cells in the mouse brain.
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DOI：
10.1016/j.molmet.2015.07.008
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发表时间：
2015-10
影响因子：
8.1
通讯作者：
Trapp S
中科院分区：
文献类型：
--
作者：
Cork SC;Richards JE;Holt MK;Gribble FM;Reimann F;Trapp S
尽管胰高血糖素样肽1是能量代谢和食物摄取的关键调节因子，但GLP-1受体的确切位置和某些人群的生理相关性仍存在争议。本研究调查了新型GLP-1R-Cre小鼠作为解决这一问题的功能工具。将在GLP1R启动子下表达Cre重组酶的小鼠与rosa26EYFP或tdRFP报告菌株杂交，以鉴定表达GLP-1R的细胞。对成年小鼠急性冠状脑片上的tdRFP阳性神经元进行膜片钳记录，通过病毒基因传递实现体内GLP-1R细胞的选择性靶向。在脑室周围器官、杏仁核、下丘脑核团和延髓腹外侧区有大量的EYFP或tdRFP免疫反应细胞。在孤束核和丘脑室旁核可见少量阳性神经元。然而，tdRFP阳性神经元也出现在没有胰高血糖素原神经元投射的区域，如海马区和皮质区。GLP-1R细胞不表达GFAP或小白蛋白，但部分细胞为儿茶酚胺能细胞。在BNST、海马和PVN的全细胞记录中证实了GLP-1R的表达，其中100nM GLP-1引发了可逆的内向电流或去极化。此外，单侧立体定向注射依赖cre的AAV到下丘脑室旁核表明tdRFP阳性细胞表达cre重组酶促进病毒介导的EYFP表达。本研究对GLP-1R在小鼠中枢神经系统的表达进行了全面的描述和表型分析。我们展示了将GLP-1R-CRE小鼠与病毒结合产生选择性分子句柄的能力，使未来能够在体内研究它们的生理重要性。这种转基因小鼠可以准确地评估表达GLP-1受体的细胞的分布。GLP-1去极化PVN、BNST和海马神经元。利用该转基因小鼠可以在体内操纵表达GLP-1R的细胞。
Although Glucagon-like peptide 1 is a key regulator of energy metabolism and food intake, the precise location of GLP-1 receptors and the physiological relevance of certain populations is debatable. This study investigated the novel GLP-1R-Cre mouse as a functional tool to address this question. Mice expressing Cre-recombinase under the Glp1r promoter were crossed with either a ROSA26 eYFP or tdRFP reporter strain to identify GLP-1R expressing cells. Patch-clamp recordings were performed on tdRFP-positive neurons in acute coronal brain slices from adult mice and selective targeting of GLP-1R cells in vivo was achieved using viral gene delivery. Large numbers of eYFP or tdRFP immunoreactive cells were found in the circumventricular organs, amygdala, hypothalamic nuclei and the ventrolateral medulla. Smaller numbers were observed in the nucleus of the solitary tract and the thalamic paraventricular nucleus. However, tdRFP positive neurons were also found in areas without preproglucagon-neuronal projections like hippocampus and cortex. GLP-1R cells were not immunoreactive for GFAP or parvalbumin although some were catecholaminergic. GLP-1R expression was confirmed in whole-cell recordings from BNST, hippocampus and PVN, where 100 nM GLP-1 elicited a reversible inward current or depolarisation. Additionally, a unilateral stereotaxic injection of a cre-dependent AAV into the PVN demonstrated that tdRFP-positive cells express cre-recombinase facilitating virally-mediated eYFP expression. This study is a comprehensive description and phenotypic analysis of GLP-1R expression in the mouse CNS. We demonstrate the power of combining the GLP-1R-CRE mouse with a virus to generate a selective molecular handle enabling future in vivo investigation as to their physiological importance. This transgenic mouse allows accurate evaluation of the distribution of GLP-1 receptor expressing cells. GLP-1 depolarises PVN, BNST and hippocampus neurons. GLP-1R expressing cells can be manipulated in vivo using this transgenic mouse.