Expression Analysis of Serotonin Receptors, Serotonin Transporter and l-Amino Acid Decarboxylase in the Mouse Sphenopalatine Ganglion by RT-PCR, Northern Blot Analysis and In Situ Hybridization
Expression Analysis of Serotonin Receptors, Serotonin Transporter and l-Amino Acid Decarboxylase in the Mouse Sphenopalatine Ganglion by RT-PCR, Northern Blot Analysis and In Situ Hybridization
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DOI：
10.1016/j.neuroscience.2019.05.028
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发表时间：
2019-07
期刊：
影响因子：
3.3
通讯作者：
Y. Ishida;Yusuke Sugiura;Takuya Magome;Takefumi Kamakura;Y. Takimoto;Yukiko Hanada;Kazuki Kitayama;Yu Nakamura;S. Shimada;N. Ohta;R. Naono-Nakayama;K. Kamijo
中科院分区：
文献类型：
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作者：
Y. Ishida;Yusuke Sugiura;Takuya Magome;Takefumi Kamakura;Y. Takimoto;Yukiko Hanada;Kazuki Kitayama;Yu Nakamura;S. Shimada;N. Ohta;R. Naono-Nakayama;K. Kamijo
蝶腭神经节（SPG）是支配鼻腺、泪腺和脑血管的副交感神经纤维的细胞体的聚集。SPG控制鼻腔分泌物、眼泪和脑血管的扩张。然而，目前尚不清楚血清素如何调节SPG功能。在这项研究中，我们研究了在小鼠SPG中参与多巴胺能系统的基因的表达。我们用RT-PCR检测了5-HT 1A、5-HT 1B、5-HT 1D、5-HT 1F、5-HT 2A、5-HT 2B、5-HT 2C、5-HT 3A、5-HT 3B、5-HT 4、5-HT 5A、5-HT 5 B、5-HT 6和5-HT 7受体以及5-羟色胺转运体、色氨酸羟化酶1和2以及L-氨基酸脱羧酶（AADC）的mRNA表达水平。RT-PCR结果显示，5-HT_3A和5-HT_3Bionotropic受体及AADC在SPG中高表达。我们接下来在SPG上进行原位杂交，在通过北方印迹验证每个cRNA探针的特异性后，在细胞水平上检查这三个基因的表达。5-HT_（3A）受体、5-HT_（3B）受体和AADC在SPG神经元胞体中的表达率分别为96.5% ± 1.0%、29.7% ± 10.7%和57.4% ± 2.9%，表明5-HT_（3A）受体几乎在所有SPG神经元中均有表达。我们的研究结果，这些关键的5-羟色胺系统基因的表达在副交感SPG提供深入了解鼻炎，结膜炎和头痛的发病机制。此外，我们的研究结果表明，靶向5-HT 3A受体可能在治疗这些疾病中具有治疗潜力。
The sphenopalatine ganglion (SPG) is a gathering of the cell bodies of parasympathetic fibers that dominate the nasal gland, lacrimal gland and cerebral blood vessels. The SPG controls nasal secretions, tears, and the dilation of cerebral blood vessels. However, it is unclear how serotonin regulates SPG functions. In this study, we investigated the expression of genes involved in the serotonergic system in the mouse SPG. We examined the mRNA expression levels of 5-HT1A, 5-HT1B, 5-HT1D, 5-HT1F, 5-HT2A, 5-HT2B, 5-HT2C, 5-HT3A, 5-HT3B, 5-HT4, 5-HT5A, 5-HT5B, 5-HT6and 5-HT7receptors, as well as serotonin transporter, tryptophan hydroxylases 1 and 2, and L-amino acid decarboxylase (AADC) by RT-PCR. It revealed that the 5-HT3Aand 5-HT3Bionotropic receptors and AADC were likely to be highly expressed in the SPG, as measured by RT-PCR. We next performedin situhybridization on the SPG to examine the expression of these three genes at the cellular level after validating the specificity of each cRNA probe by northern blotting. The 5-HT3Areceptor, 5-HT3Breceptor, and AADC were expressed in 96.5% ± 1.0%, 29.7% ± 10.7%, and 57.4% ± 2.9% of neuronal cell bodies in the SPG, respectively, indicating that the 5-HT3Areceptor was virtually expressed in all SPG neurons. Our results on the expression of these critical serotonin system genes in the parasympathetic SPG provide insight into the pathogenetics of rhinitis, conjunctivitis and headache. Furthermore, our findings suggest that targeting the 5-HT3Areceptor might have therapeutic potential in the treatment of these ailments.