Human olfactory epithelial cells generated in vitro express diverse neuronal characteristics.
Human olfactory epithelial cells generated in vitro express diverse neuronal characteristics.
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DOI：
10.1016/j.neuroscience.2008.09.059
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发表时间：
2009-01-23
期刊：
影响因子：
3.3
通讯作者：
Hahn CG
中科院分区：
文献类型：
--
作者：
Borgmann-Winter KE;Rawson NE;Wang HY;Wang H;Macdonald ML;Ozdener MH;Yee KK;Gomez G;Xu J;Bryant B;Adamek G;Mirza N;Pribitkin E;Hahn CG
嗅觉上皮是从活人身上获得的再生神经细胞的唯一来源。因此，源自嗅觉上皮活检的原代培养物可用于研究不同条件和疾病状态下个体的神经生物学特征。在这里，利用这种培养物，我们报告了在体外产生的细胞，这些细胞表现出复杂的神经元表型，包括与嗅觉和中枢神经系统功能其他方面相关的受体和信号通路。利用原位杂交，我们首次证明了嗅觉上皮组织培养细胞中嗅觉受体的天然表达。我们使用免疫细胞化学、钙成像和分子方法进一步确定这些细胞中嗅觉转导分子的存在和功能。蛋白质印迹分析显示多巴胺 (D2R)、血清素 (5HT2C) 和 NMDA 亚型 1 和 2A/2B 神经递质受体的表达。基于 35S-GTP 掺入测定，多巴胺或血清素刺激以亚型特异性方式增强受体 G 蛋白激活。通过 NMDAR 2A/2B 酪氨酸磷酸化的增强以及响应 NMDA 刺激而招募信号传导伙伴，证明了培养细胞的功能特征。这里观察到的一系列神经元特征证实，源自人类嗅上皮的增殖细胞在体外分化以表达成熟嗅觉神经元的功能和分子属性。这些培养的神经细胞表现出在许多神经精神疾病中重要的神经递质途径。因此，它们可以从活人中随时获得，从而提供了一种新工具，可以将神经细胞的功能和分子特征与个体活体患者的临床特征联系起来。
The olfactory epithelium constitutes the sole source of regenerating neural cells that can be obtained from a living human. As such, primary cultures derived from olfactory epithelial biopsies can be utilized to study neurobiological characteristics of individuals under different conditions and disease states. Here, using such cultures, we report in vitro generation of cells that exhibit a complex neuronal phenotype, encompassing receptors and signaling pathways pertinent to both olfaction and other aspects of CNS function. Using in situ hybridization, we demonstrate for the first time the native expression of olfactory receptors in cultured cells derived from olfactory epithelial tissue. We further establish the presence and function of olfactory transduction molecules in these cells using immunocytochemistry, calcium imaging and molecular methods. Western blot analysis revealed the expression of neurotransmitter receptors for dopamine (D2R), serotonin (5HT2C) and NMDA subtypes 1 and 2A/2B. Stimulation with dopamine or serotonin enhanced receptor G protein activation in a subtype specific manner, based on 35S-GTP incorporation assay. Functional characteristics of the cultured cells are demonstrated through enhanced tyrosine phosphorylation of NMDAR 2A/2B and recruitment of signaling partners in response to NMDA stimulation. The array of neuronal characteristics observed here establish that proliferating cells derived from the human olfactory epithelium differentiate in vitro to express functional and molecular attributes of mature olfactory neurons. These cultured neural cells exhibit neurotransmitter pathways important in a number of neuropsychiatric disorders. Their ready availability from living humans thus provides a new tool to link functional and molecular features of neural cells with clinical characteristics of individual living patients.
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影响因子：
17.7
作者：
Hahn, CG;Gomez, G;Rawson, NE
通讯作者：
Rawson, NE
影响因子：
3.3
作者：
MANIAFARNELL, BL;FARBMAN, AI;BRUCH, RC
通讯作者：
BRUCH, RC
影响因子：
2.9
作者：
Okada, Y;Miyamoto, T;Toda, K
通讯作者：
Toda, K
影响因子：
2.5
作者：
Friedman, D;Strowbridge, BW
通讯作者：
Strowbridge, BW
影响因子：
7.8
作者：
Ghanbari, HA;Ghanbari, K;Perry, G
通讯作者：
Perry, G