大鼠嗅球气味响应在体分析与生物电子鼻研究
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摘要
生物嗅觉系统对气味具有较高的辨识度,在气味识别速度、气味识别精度上远远超过目前基于化学传感器阵列构建的电子鼻系统。但如何将生物嗅觉应用于实际的气味检测中,仍然存在着很大的挑战。气味分子在动物鼻腔中与嗅觉感受细胞作用后,化学信息转变成生物电信号传导至嗅觉信息处理的中转站—嗅球。通过嗅球对气味信息的处理整合,嗅觉信息进一步传递给大脑皮层中嗅觉相关的皮层,实现气味认知。其中,嗅球被认为是嗅觉初级中枢,对嗅觉形成起到了关键的作用。理解嗅球的气味认知机制,在嗅觉信息处理和实际气味检测中都具有重要意义。
目前,对于气味刺激产生的嗅球细胞响应记录主要采用膜片钳技术和光学成像技术。膜片钳技术不能实现多点同步测量,缺乏大量神经细胞协同作用的信息,而光学成像技术背景噪声大,成像精度仍较难突破。因此,采用植入式微电极阵列传感器对嗅球电生理信号进行长时程、多点同步的监测与分析,有利于理解嗅球细胞群的信息处理,实现生物电子鼻的设计。本文对嗅觉系统多个层次的细胞进行了基于电生理数据的细胞建模与仿真,设计了基于植入式传感器的在体嗅球僧帽细胞气味刺激响应记录与分析平台,分析了嗅球僧帽细胞群气味响应模式。结合生物学和工程学手段,构建新型的生物电子鼻系统实现气味识别。
本论文的主要内容和贡献如下：
1.对嗅觉系统多个层次的细胞进行建模仿真。利用电生理信号建立了基于电压门控通道的嗅觉感受细胞电生理模型,并仿真了嗅觉感受细胞受到刺激时引起细胞动作电位的发放。仿真了僧帽细胞电生理模型。探索了嗅觉系统模型对嗅觉生理现象的仿真,并结合生理实验测得的数据对模型参数进行了优化。
2.设计了麻醉大鼠嗅觉气味响应研究系统。该系统包括气味刺激装置、呼吸信号记录装置、电生理信号记录装置、记录电极的制备平台和数据分析平台。详细描述了大鼠的手术流程与电极植入过程,并对嗅球切片进行染色,验证电极植入位置。
3.提出了短时程气味刺激下大鼠的快速气味感知,实现了时间依赖性的气味分类。采用细胞群脉冲发放矩阵模式分析法,在特征空间中绘制了僧帽细胞群对气味刺激的空间响应曲线,表明麻醉大鼠在短时程气味刺激后的第一个呼吸周期内就完成了对气味的感知。选取细胞发放相似时间段内同步记录的僧帽细胞群脉冲发放特征,用主成分分析方法实现了对五种气味的分类。
4.提出基于僧帽细胞层场电位信号的生物电子鼻设计。低频场电位信号稳定且容易获取,在不同气味刺激下功率谱能量分布存在差异。使用K最邻近分类算法对多窗法谱估计得到的四种气味刺激时频图进行分类,在信号伽马频段(40-120Hz)得到77.4%的分类准确率。
Olfactory system has exquisite capability to discriminate odors, especially in speed and accuracy of odor recognition, which is much better than current electronic nose based on chemical sensor array. However, there are still many challenges remained in applying biological olfaction to industrial odor detection. Odor molecules interact with olfactory receptor neurons (ORNs) in nasal nostrils. During the process, chemical signals are changed into electric signals, and transmitted to the relay station of odor information processing called olfactory bulb (OB). After coding process of odor information in OB, the olfactory cortex receives the signal from OB and perceives odors. OB is considered as primary cortex of olfaction, which plays an important role in odor perception. Understanding the mechanism of odor perception in OB is worthwhile for both odor information processing and industrial odor detection.
Currently, patch clamp recording technique and optical imaging technique are primary measurement methods for odor-evoked response recording of OB cells. Constrained by recording few cells synchronously, patch clamp technique cannot provide coherent activities of large numbers of neurons. While optical imaging technique has barriers in image resolution due to background noise. Therefore, with the advantage of long-term, synchronous multi-site measurement, implanted microelectrode arrays have been used for electrophysiological recording of OB. This technique benefits for understanding of cell information processing, furthermore, helps for accomplishing design of bio-enose. Based on the electrophysiological recordings, the paper established and simulated multiple models of olfactory cells and network including ORN, mitral cell, and olfactory neuronal network. In addition, the paper described the platform built for odor stimulation and odor-evoked response recording of OB cells in vivo based on implanted sensors. Using this platform, We investigated the temporal firing patterns of mitral cell ensemble. Combining biological method with engineering method, we established bio-enose system for odor recognition.
The major contents and contributions of this thesis are as follows:
1. Modeling and simulating neurons and neuron network of olfactory system. Based on electrophysiological data of voltage-gated channels, we established ORN model and simulated the action potential evoked by current pulse stimulation. In addition, we simulated the model of mitral cell and neuron network of olfactory system, and optimized parameters in models according to experimental data.
2. Establishing research platform for study on odor-evoked responses of anesthetized free-breathing rats. The platform included odor delivery module, respiratory signal recording module, electrophysiological signal recording module, electrode fabrication module and data analysis module. Procedures of surgery and electrode insertion were illustrated in detail. The recording site of electrode array was examined by post-mortem histological reconstruction of the electrode track in OB slice.
3. Uncovering rapid odor perception of anesthetized free-breathing rat in OB under short-term odor stimulation and context-based odor discrimination by mitral cell group. Utilizing matrix paradigm of cell group firing, we drew response curve of mitral cell ensemble in feature space which indicates that odor perception was achieved within the first breathing cycle after short-term odor stimulation. Features of odor-evoked response were extracted from mitral cells which were synchronously recorded with similar firing rate histogram. The result showed that five odors can be classified by principal component analysis.
4. Designing bioelectronic nose system based on field potential of mitral cell layer in OB. Field potential is stable and easy to acquire. Besides, power spectrum distributions have difference under different odor stimuli. We applied K-nearest neighboring classification method for spectrograms of four odors stimuli by multitaper spectrum estimation. The classification accuracy of four odors reached77.4%in gamma band (40-120Hz).
目前,对于气味刺激产生的嗅球细胞响应记录主要采用膜片钳技术和光学成像技术。膜片钳技术不能实现多点同步测量,缺乏大量神经细胞协同作用的信息,而光学成像技术背景噪声大,成像精度仍较难突破。因此,采用植入式微电极阵列传感器对嗅球电生理信号进行长时程、多点同步的监测与分析,有利于理解嗅球细胞群的信息处理,实现生物电子鼻的设计。本文对嗅觉系统多个层次的细胞进行了基于电生理数据的细胞建模与仿真,设计了基于植入式传感器的在体嗅球僧帽细胞气味刺激响应记录与分析平台,分析了嗅球僧帽细胞群气味响应模式。结合生物学和工程学手段,构建新型的生物电子鼻系统实现气味识别。
本论文的主要内容和贡献如下：
1.对嗅觉系统多个层次的细胞进行建模仿真。利用电生理信号建立了基于电压门控通道的嗅觉感受细胞电生理模型,并仿真了嗅觉感受细胞受到刺激时引起细胞动作电位的发放。仿真了僧帽细胞电生理模型。探索了嗅觉系统模型对嗅觉生理现象的仿真,并结合生理实验测得的数据对模型参数进行了优化。
2.设计了麻醉大鼠嗅觉气味响应研究系统。该系统包括气味刺激装置、呼吸信号记录装置、电生理信号记录装置、记录电极的制备平台和数据分析平台。详细描述了大鼠的手术流程与电极植入过程,并对嗅球切片进行染色,验证电极植入位置。
3.提出了短时程气味刺激下大鼠的快速气味感知,实现了时间依赖性的气味分类。采用细胞群脉冲发放矩阵模式分析法,在特征空间中绘制了僧帽细胞群对气味刺激的空间响应曲线,表明麻醉大鼠在短时程气味刺激后的第一个呼吸周期内就完成了对气味的感知。选取细胞发放相似时间段内同步记录的僧帽细胞群脉冲发放特征,用主成分分析方法实现了对五种气味的分类。
4.提出基于僧帽细胞层场电位信号的生物电子鼻设计。低频场电位信号稳定且容易获取,在不同气味刺激下功率谱能量分布存在差异。使用K最邻近分类算法对多窗法谱估计得到的四种气味刺激时频图进行分类,在信号伽马频段(40-120Hz)得到77.4%的分类准确率。
Olfactory system has exquisite capability to discriminate odors, especially in speed and accuracy of odor recognition, which is much better than current electronic nose based on chemical sensor array. However, there are still many challenges remained in applying biological olfaction to industrial odor detection. Odor molecules interact with olfactory receptor neurons (ORNs) in nasal nostrils. During the process, chemical signals are changed into electric signals, and transmitted to the relay station of odor information processing called olfactory bulb (OB). After coding process of odor information in OB, the olfactory cortex receives the signal from OB and perceives odors. OB is considered as primary cortex of olfaction, which plays an important role in odor perception. Understanding the mechanism of odor perception in OB is worthwhile for both odor information processing and industrial odor detection.
Currently, patch clamp recording technique and optical imaging technique are primary measurement methods for odor-evoked response recording of OB cells. Constrained by recording few cells synchronously, patch clamp technique cannot provide coherent activities of large numbers of neurons. While optical imaging technique has barriers in image resolution due to background noise. Therefore, with the advantage of long-term, synchronous multi-site measurement, implanted microelectrode arrays have been used for electrophysiological recording of OB. This technique benefits for understanding of cell information processing, furthermore, helps for accomplishing design of bio-enose. Based on the electrophysiological recordings, the paper established and simulated multiple models of olfactory cells and network including ORN, mitral cell, and olfactory neuronal network. In addition, the paper described the platform built for odor stimulation and odor-evoked response recording of OB cells in vivo based on implanted sensors. Using this platform, We investigated the temporal firing patterns of mitral cell ensemble. Combining biological method with engineering method, we established bio-enose system for odor recognition.
The major contents and contributions of this thesis are as follows:
1. Modeling and simulating neurons and neuron network of olfactory system. Based on electrophysiological data of voltage-gated channels, we established ORN model and simulated the action potential evoked by current pulse stimulation. In addition, we simulated the model of mitral cell and neuron network of olfactory system, and optimized parameters in models according to experimental data.
2. Establishing research platform for study on odor-evoked responses of anesthetized free-breathing rats. The platform included odor delivery module, respiratory signal recording module, electrophysiological signal recording module, electrode fabrication module and data analysis module. Procedures of surgery and electrode insertion were illustrated in detail. The recording site of electrode array was examined by post-mortem histological reconstruction of the electrode track in OB slice.
3. Uncovering rapid odor perception of anesthetized free-breathing rat in OB under short-term odor stimulation and context-based odor discrimination by mitral cell group. Utilizing matrix paradigm of cell group firing, we drew response curve of mitral cell ensemble in feature space which indicates that odor perception was achieved within the first breathing cycle after short-term odor stimulation. Features of odor-evoked response were extracted from mitral cells which were synchronously recorded with similar firing rate histogram. The result showed that five odors can be classified by principal component analysis.
4. Designing bioelectronic nose system based on field potential of mitral cell layer in OB. Field potential is stable and easy to acquire. Besides, power spectrum distributions have difference under different odor stimuli. We applied K-nearest neighboring classification method for spectrograms of four odors stimuli by multitaper spectrum estimation. The classification accuracy of four odors reached77.4%in gamma band (40-120Hz).
引文
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