Elucidating molecular mechanisms mediating cellular responses to touch and pain

阐明介导细胞对触摸和疼痛反应的分子机制

基本信息

批准号：
9185225
负责人：
Elizabeth A. Heath-Heckman
金额：
$ 5.61万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-12-01 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9185225
关键词：
Action Potentials Address Affect Afferent Neurons Animals Behavior Biological Assay Biological Models CRISPR/Cas technology Calcium Cell Culture System Cell Culture Techniques Cells Characteristics Collaborations Cues Data Deltastab Development Disease Embryo Environment Exhibits Ganglia Genes Genetic Transcription Hand Head Health Human In Situ Hybridization Invertebrates Knock-out Lead Leeches Length Life Mammalian Cell Measurement Measures Mechanics Mediating Modeling Molecular Nature Nerve Nervous system structure Neurobiology Neurons Neurophysiology - biologic function Pain Pattern Plants Plasmids Play Preparation Process Prokaryotic Cells Proteins Quantitative Reverse Transcriptase PCR RNA Interference Reproducibility Response to stimulus physiology Role Sensory Ganglia Signal Transduction Stimulus System Techniques Touch sensation Transcript Transducers Withdrawal candidate validation cell type chronic itch chronic pain differential expression experimental study expression vector gene function genome editing imaging potential improved in vivo insight interest knock-down loss of function mechanical force mechanotransduction neuron development public health relevance response transcriptome transcriptome sequencing transcriptomics

项目摘要

DESCRIPTION (provided by applicant): Mechanotransduction, or the mechanisms by which cells convert mechanical stimuli into electrical activity, is a process conserved across all domains of life and is a necessary facet of our interaction with the world. However, despite its ubiquity and importance, mechanotransduction is not well understood at a molecular level. Leeches in the genus Hirudo, commonly known as medicinal leeches, have been long used as a model system for neurobiology and as a result the characteristics of the neurons in the segmentally iterated ganglia of the ventral nerve cord have been well established. To determine what genes might mediate the differences between the mechanosensitive and non-mechanosensitive neurons in these ganglia, I and others carried out a transcriptomic analysis of five different cell types in the Hirudo ganglion. One major signature that arose from this analysis was the differential expression of several transcripts encoding channels similar to those that have been shown to be mechanosensitive in other systems. However, our system would not only allow us to ground- truth whether these channels confer mechanosensitivity, but also would determine the nature of this process, as the three different mechanosensitive neurons in the leech ganglion exhibit different responses to touch. Specifically, I plan to: 1) Determine the effect of the loss of these channels on neuronal function by transcript knock-down in Hirudo body wall preparations and in vivo neuronal culture; 2) Describe the developmental trajectory and organismal relevance of these genes using Helobdella, another leech and well-established developmental model system; and 3) Directly determine whether a subset of these channels confers mechanosensitivity in mammalian cell culture through expression of the full-length protein and subsequent functional assays.

描述（由申请人提供）：机械转导，或细胞将机械刺激转化为电活动的机制，是生命所有领域中保守的过程，是我们与世界互动的一个必要方面，然而，尽管它普遍存在且很重要。水蛭属中的机械传导在分子水平上尚未得到很好的理解，通常被称为药用水蛭，长期以来一直被用作神经生物学的模型系统，因此为了确定哪些基因可能介导这些神经节中机械敏感神经元和非机械敏感神经元之间的差异，我和其他人对五个不同的神经节进行了转录组分析。水蛭神经节中的细胞类型是这一分析产生的一个主要特征。是几个转录本编码通道的差异表达，这些通道与其他系统中已被证明具有机械敏感性的通道相似。然而，我们的系统不仅可以让我们了解这些通道是否具有机械敏感性，而且还可以确定其性质。具体来说，我计划：1）通过水蛭体内的转录本敲低来确定这些通道的丢失对神经功能的影响。壁制备和体内神经培养；2) 使用 Helobdella（另一种水蛭和成熟的发育模型系统）描述这些基因的发育轨迹和有机相关性；3) 直接确定这些通道的子集是否在哺乳动物细胞培养中赋予机械敏感性。通过全长蛋白质的表达和随后的功能测定。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Behavioral analysis of substrate texture preference in a leech, Helobdella austinensis.

水蛭基质纹理偏好的行为分析，Helobdella austinensis。

DOI：
发表时间：
2019
期刊：
影响因子：
0
作者：
Kim, Rachel C;Le, Dylan;Ma, Kenny;Heath;Whitehorn, Nathan;Kristan Jr, William B;Weisblat, David A
通讯作者：
Weisblat, David A

The Metronome of Symbiosis: Interactions Between Microbes and the Host Circadian Clock.

共生节拍器：微生物与宿主昼夜节律时钟之间的相互作用。

DOI：
发表时间：
2016
期刊：
影响因子：
2.6
作者：
Heath
通讯作者：
Heath

Transcriptional profiling of identified neurons in leech.

水蛭中已识别神经元的转录谱。

DOI：
发表时间：
2021-03-25
期刊：
影响因子：
4.4
作者：
Heath;Yoo, Shinja;Winchell, Christopher;Pellegrino, Maurizio;Angstadt, James;Lammardo, Veronica B;Bautista, Diana;De;Weisblat, David
通讯作者：
Weisblat, David