In vivo Optical Access to Olfactory Sensory Neurons in the Mouse Olfactory Epithelium
In vivo Optical Access to Olfactory Sensory Neurons in the Mouse Olfactory Epithelium
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DOI：
10.21769/bioprotoc.4055
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发表时间：
2021-06-20
期刊：
影响因子：
0.8
通讯作者：
Imai, Takeshi
中科院分区：
文献类型：
--
作者：
Inagaki, Shigenori;Iwata, Ryo;Imai, Takeshi
在神经科学领域，理解感觉刺激在感觉系统的入口处如何转化为神经活动至关重要。在嗅觉系统中，吸入鼻腔的气味分子由约1000种气味受体（ORs）检测，这些受体由嗅觉感觉神经元（OSNs）表达。由于每个嗅觉感觉神经元仅表达一种气味受体，气味诱发的反应反映了气味分子与所表达的气味受体之间的相互作用。嗅觉感觉神经元胞体的反应通常通过钙成像和电生理技术来测量；然而，以往的技术需要进行组织解剖或细胞解离，这使得研究生理反应颇具难度。在此，我们介绍一种方案，该方案使我们能够在活体状态下观察小鼠嗅上皮中单个嗅觉感觉神经元胞体的气味诱发反应。通过薄化头骨进行双光子激发，利用基因编码的钙指示剂GCaMP实现高灵敏度的钙成像。记录自由呼吸小鼠嗅觉感觉神经元胞体的气味诱发反应，对于理解气味信息在周边及大脑高级神经回路中的处理方式具有重要意义。
In neuroscience, it is fundamental to understand how sensory stimuli are translated into neural activity at the entry point of sensory systems. In the olfactory system, odorants inhaled into the nasal cavity are detected by similar to 1,000 types of odorant receptors (ORs) that are expressed by olfactory sensory neurons (OSNs). Since each OSN expresses only one type of odorant receptor, the odorevoked responses reflect the interaction between odorants and the expressed OR. The responses of OSN somata are often measured by calcium imaging and electrophysiological techniques; however, previous techniques require tissue dissection or cell dissociation, rendering it difficult to investigate physiological responses. Here, we describe a protocol that allows us to observe odor-evoked responses of individual OSN somata in the mouse olfactory epithelium in vivo. Two-photon excitation through the thinned skull enables highly-sensitive calcium imaging using a genetically encoded calcium indicator, GCaMP. Recording of odor-evoked responses in OSN somata in freely breathing mice will be fundamental to understanding how odor information is processed at the periphery and higher circuits in the brain.