Rau Andrew R, Hughes Alexander R, Hentges Shane T
Department of Biomedical Sciences, Colorado State University , Fort Collins, Colorado.
Am J Physiol Regul Integr Comp Physiol. 2018 Jul 1;315(1):R144-R152. doi: 10.1152/ajpregu.00047.2018. Epub 2018 Mar 28.
Products of the proopiomelanocortin (POMC) prohormone regulate aspects of analgesia, reward, and energy balance; thus, the neurons that produce POMC in the hypothalamus have received considerable attention. However, there are also cells in the nucleus of the solitary tract (NTS) that transcribe Pomc, although low levels of Pomc mRNA and relative lack of POMC peptide products in the adult mouse NTS have hindered the study of these cells. Therefore, studies of NTS POMC cells have largely relied on transgenic mouse lines. Here, we set out to determine the amino acid (AA) transmitter phenotype of NTS POMC neurons by using Pomc-Gfp transgenic mice to identify POMC cells. We found that cells expressing the green fluorescent protein (GFP) represent a mix of GABAergic and glutamatergic cells as indicated by Gad2 and vesicular Glut2 ( vGlut2) mRNA expression, respectively. We then examined the AA phenotype of POMC cells labeled by a Pomc-Cre transgene and found that these are also a mix of GABAergic and glutamatergic cells. However, the NTS cells labeled by the Gfp- and Cre-containing transgenes represented distinct populations of cells in three different Pomc-Cre mouse lines. Consistent with previous work, we were unable to reliably detect Pomc mRNA in the NTS despite clear expression in the hypothalamus. Thus, it was not possible to determine which transgenic tool most accurately identifies NTS cells that may express Pomc or release POMC peptides, although the results indicate the transgenic tools for study of these NTS neurons can label disparate populations of cells with varied AA phenotypes.
阿片促黑激素皮质素原(POMC)前体激素的产物调节镇痛、奖赏和能量平衡等方面;因此,下丘脑产生POMC的神经元受到了广泛关注。然而,孤束核(NTS)中也有细胞转录Pomc,尽管成年小鼠NTS中Pomc mRNA水平较低且POMC肽产物相对缺乏,这阻碍了对这些细胞的研究。因此,对NTS POMC细胞的研究很大程度上依赖于转基因小鼠品系。在这里,我们通过使用Pomc-Gfp转基因小鼠来鉴定POMC细胞,以确定NTS POMC神经元的氨基酸(AA)递质表型。我们发现,表达绿色荧光蛋白(GFP)的细胞分别由Gad2和囊泡型谷氨酸转运体2(vGlut2)mRNA表达表明为GABA能和谷氨酸能细胞的混合。然后,我们检查了由Pomc-Cre转基因标记的POMC细胞的AA表型,发现这些细胞也是GABA能和谷氨酸能细胞的混合。然而,在三种不同的Pomc-Cre小鼠品系中,由含Gfp和Cre的转基因标记的NTS细胞代表了不同的细胞群体。与之前的工作一致,尽管下丘脑有明显表达,但我们无法在NTS中可靠地检测到Pomc mRNA。因此,尽管结果表明用于研究这些NTS神经元的转基因工具可以标记具有不同AA表型的不同细胞群体,但无法确定哪种转基因工具最准确地识别可能表达Pomc或释放POMC肽的NTS细胞。