Bryche Bertrand, Saint-Albin Audrey, Le Poupon Schlegel Claire, Baly Christine, Congar Patrice, Meunier Nicolas
Neurobiologie de l'olfaction, Institut National de Recherche Agronomique (INRA), Université Paris-Saclay, Jouy-en-Josas; Université de Versailles Saint-Quentin en Yvelines, Département de Biologie, Versailles, France.
Neurobiologie de l'olfaction, Institut National de Recherche Agronomique (INRA), Université Paris-Saclay, Jouy-en-Josas, France.
Neural Regen Res. 2020 Feb;15(2):352-360. doi: 10.4103/1673-5374.265558.
The olfactory mucosa holds olfactory sensory neurons directly in contact with an aggressive environment. In order to maintain its integrity, it is one of the few neural zones which are continuously renewed during the whole animal life. Among several factors regulating this renewal, endothelin acts as an anti-apoptotic factor in the rat olfactory epithelium. In the present study, we explored whether endothelin could also act as a proliferative factor. Using primary culture of the olfactory mucosa, we found that an early treatment with endothelin increased its growth. Consistently, a treatment with a mixture of BQ and BQ (endothelin receptor antagonists) decreased the primary culture growth without affecting the cellular death level. We then used combined approaches of calcium imaging, reverse transcriptase-quantitative polymerase chain reaction and protein level measurements to show that endothelin was locally synthetized by the primary culture until it reached confluency. Furthermore, in vivo intranasal instillation of endothelin receptor antagonists led to a decrease of olfactory mucosa cell expressing proliferating cell nuclear antigen (PCNA), a marker of proliferation. Only short-term treatment reduced the PCNA level in the olfactory mucosa cells. When the treatment was prolonged, the PCNA level was not statistically affected but the expression level of endothelin was increased. Overall, our results show that endothelin plays a proliferative role in the olfactory mucosa and that its level is dynamically regulated. This study was approved by the Comité d'éthique en expérimentation animale COMETHEA (COMETHEA C2EA -45; protocol approval #12-058) on November 28, 2012.
嗅觉黏膜使嗅觉感觉神经元直接与具有侵袭性的环境接触。为了维持其完整性，它是整个动物生命过程中少数持续更新的神经区域之一。在调节这种更新的几个因素中，内皮素在大鼠嗅觉上皮中作为一种抗凋亡因子发挥作用。在本研究中，我们探讨了内皮素是否也能作为一种增殖因子。通过嗅觉黏膜的原代培养，我们发现早期用内皮素处理可增加其生长。一致地，用BQ和BQ（内皮素受体拮抗剂）混合物处理会降低原代培养的生长，而不影响细胞死亡水平。然后，我们使用钙成像、逆转录定量聚合酶链反应和蛋白质水平测量等联合方法，以表明内皮素在原代培养达到汇合之前是在局部合成的。此外，体内鼻内滴注内皮素受体拮抗剂导致表达增殖细胞核抗原（PCNA，一种增殖标志物）的嗅觉黏膜细胞减少。只有短期处理会降低嗅觉黏膜细胞中的PCNA水平。当处理延长时，PCNA水平在统计学上没有受到影响，但内皮素的表达水平增加。总体而言，我们的结果表明内皮素在嗅觉黏膜中发挥增殖作用，并且其水平受到动态调节。本研究于2012年11月28日获得动物实验伦理委员会COMETHEA（COMETHEA C2EA -45；方案批准号#12-058）的批准。
