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首页> 外文期刊>Cilia >Assaying sensory ciliopathies using calcium biosensor expression in zebrafish ciliated olfactory neurons
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Assaying sensory ciliopathies using calcium biosensor expression in zebrafish ciliated olfactory neurons

机译：使用钙生物传感器表达的斑马鱼纤毛嗅觉神经元检测感觉纤毛病。

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Primary cilia mediate signal transduction by acting as an organizing scaffold for receptors, signalling proteins and ion channels. Ciliated olfactory sensory neurons (OSNs) organize olfactory receptors and ion channels on cilia and generate a calcium influx as a primary signal in odourant detection. In the zebrafish olfactory placode, ciliated OSNs and microvillus OSNs constitute the major OSN cell types with distinct odourant sensitivity. Using transgenic expression of the calcium biosensor GCaMP5 in OSNs, we analysed sensory cilia-dependent odour responses in live zebrafish, at individual cell resolution. oval/ift88 mutant and ift172 knockdown zebrafish were compared with wild-type siblings to establish ciliated OSN sensitivity to different classes of odourants. oval/ift88 mutant and ift172 knockdown zebrafish showed fewer and severely shortened OSN cilia without a reduction in OSN number. The fraction of responding OSNs and response amplitudes to bile acids and food odour, both sensed by ciliated OSNs, were significantly reduced in ift88 mutants and ift172-deficient embryos, while the amino acids responses were not significantly changed. Our approach presents a quantitative model for studying sensory cilia signalling using zebrafish OSNs. Our results also implicate ift172-deficiency as a novel cause of hyposmia, a reduced sense of smell, highlighting the value of directly assaying sensory cilia signalling in vivo and supporting the idea that hyposmia can be used as a diagnostic indicator of ciliopathies.
机译：原发纤毛通过充当受体，信号蛋白和离子通道的组织支架来介导信号转导。纤毛嗅觉感觉神经元（OSN）在纤毛上组织嗅觉受体和离子通道，并产生钙流入作为气味检测中的主要信号。在斑马鱼的嗅觉平台中，纤毛OSN和微绒毛OSN构成主要的OSN细胞类型，具有独特的气味敏感性。使用OSNs中钙生物传感器GCaMP5的转基因表达，我们在单个细胞分辨率下分析了活斑马鱼中感官纤毛依赖性气味反应。将椭圆形/ ift88突变体和ift172连锁的斑马鱼与野生型兄弟姐妹进行比较，以建立纤毛OSN对不同种类气味的敏感性。椭圆形/ ift88突变体和ift172连锁的斑马鱼显示出更少且严重缩短的OSN纤毛，而OSN数量却没有减少。纤毛状OSN感测到的响应OSN分数和对胆汁酸和食物异味的响应幅度在ift88突变体和ift172缺陷型胚胎中均显着降低，而氨基酸响应没有明显改变。我们的方法提出了一种定量模型，用于研究使用斑马鱼OSN的感觉纤毛信号。我们的研究结果还暗示ift172缺乏症是低渗症的一种新原因，一种嗅觉降低的现象，突显了直接在体内检​​测感官纤毛信号的价值，并支持了低渗症可以用作纤毛病的诊断指标的观点。

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