A novel bidirectional interaction between endothelin-3 and retinoic acid in rat enteric nervous system precursors.
A novel bidirectional interaction between endothelin-3 and retinoic acid in rat enteric nervous system precursors.
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DOI：
10.1371/journal.pone.0074311
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Gariepy CE
中科院分区：
文献类型：
--
作者：
Gisser JM;Cohen AR;Yin H;Gariepy CE
通过内皮素受体B(EDNRB)传递的信号对肠道神经系统(ENS)的发育至关重要，内皮素系统基因的突变导致人类先天性巨结肠无神经节细胞增多症。疾病的外显性受到其他遗传因素的调节。影响维甲酸(RA)信号的突变也会导致小鼠神经节细胞缺乏症。因此，我们假设RA和内皮素信号通路可能在控制ENS的发展中相互作用。免疫选择的大鼠ENS前体细胞与EDNRB配体内皮素-3、EDNRB选择性拮抗剂(BQ-788)和/或RA共同培养3或14天。3天后检测ENS发育相关基因、RA和EDNRB信号相关基因的mRNA水平。对增殖细胞和表达神经元、神经胶质细胞和肌成纤维细胞标志物的细胞进行量化。分离的ENS前体细胞与RA共同培养3天后，ET-3基因及其激活酶的表达降低。与不含RA的培养相比，这些变化与胶质细胞增殖、胶质细胞比例较高以及神经元比例较低有关。这些变化与EDNRB信号无关。相反，在这些培养物中激活EDNRB会降低RA受体β和γ的表达，并影响RA合成酶和降解酶的表达。这些基因表达的变化与培养中胶质细胞增殖减少和胶质细胞比例降低有关。在没有EDNRB信号的情况下，14天后，RA诱导形成一个充满神经节、神经胶质细胞和肌成纤维细胞的异细胞神经丛。在体外，存在复杂的内皮素-维甲酸相互作用，协调调节大鼠ENS前体的发育。这些结果表明，环境RA可能调节内皮素突变个体的无神经节细胞增多症的表达。
Signaling through the endothelin receptor B (EDNRB) is critical for the development of the enteric nervous system (ENS) and mutations in endothelin system genes cause Hirschsprung’s aganglionosis in humans. Penetrance of the disease is modulated by other genetic factors. Mutations affecting retinoic acid (RA) signaling also produce aganglionosis in mice. Thus, we hypothesized that RA and endothelin signaling pathways may interact in controlling development of the ENS. Rat immunoselected ENS precursor cells were cultured with the EDNRB ligand endothelin-3, an EDNRB-selective antagonist (BQ-788), and/or RA for 3 or 14 days. mRNA levels of genes related to ENS development, RA- and EDNRB-signaling were measured at 3 days. Proliferating cells and cells expressing neuronal, glial, and myofibroblast markers were quantified. Culture of isolated ENS precursors for 3 days with RA decreases expression of the endothelin-3 gene and that of its activation enzyme. These changes are associated with glial proliferation, a higher percentage of glia, and a lower percentage of neurons compared to cultures without RA. These changes are independent of EDNRB signaling. Conversely, EDNRB activation in these cultures decreases expression of RA receptors β and γ mRNA and affects the expression of the RA synthetic and degradative enzymes. These gene expression changes are associated with reduced glial proliferation and a lower percentage of glia in the culture. Over 14 days in the absence of EDNRB signaling, RA induces the formation of a heterocellular plexus replete with ganglia, glia and myofibroblasts. A complex endothelin-RA interaction exists that coordinately regulates the development of rat ENS precursors in vitro. These results suggest that environmental RA may modulate the expression of aganglionosis in individuals with endothelin mutations.