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Arabidopsis thaliana

ABSTRACT: Mechanosensory Trichome Cells Evoke Mechanical Stimuli-induced Immune Response in Plants

PROVIDER: PRJDB11645 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	DRR290976.fastq.gz	Fastqsanger.gz
	DRR290977.fastq.gz	Fastqsanger.gz
	DRR290978.fastq.gz	Fastqsanger.gz
	DRR290979.fastq.gz	Fastqsanger.gz
	DRR290980.fastq.gz	Fastqsanger.gz

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Project description:Trichomes of Arabidopsis thaliana have been broadly used to study cell development, cell differentiation and cell wall biogenesis. In this context, the exposed position, extraordinary size and characteristic morphology of trichomes featured particularly the exploration of trichome mutant phenotypes. However, trichome-specific biochemical or -omics analyses require a proper separation of trichomes from residual plant tissue. Thus, different strategies were proposed in the past for trichome isolation, which mostly rely on harsh conditions. To improve trichome-leaf separation, we revised a previously proposed method for isolating A. thaliana trichomes by optimizing the mechanical and biochemical specifications for trichome release. Furthermore, we introduced a density gradient centrifugation step to remove residual plant debris. We found that prolonged, yet mild seedling agitation increases the overall trichome yield by about 62% compared to the original protocol. We noticed that subsequent density gradient centrifugation further visually enhances trichome purity, which could be advantageous for downstream analyses. Histochemical and biochemical investigation of trichome cell wall composition indicated that gentle agitation during trichome release largely preserves trichome integrity. We used enriched and purified trichomes for proteomic analysis and present a reference data set of trichome-resident and -enriched proteins.

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Project description:The sense of touch relies on the detection of mechanical stimuli by specialized sensory neurons. Mechanosensory neurons are generated from neural crest cells, but the scarcity of molecular data has made it difficult to analyze their development and to define the basis of their diversity and function. The transcription factor c-Maf is crucial for mechanosensory function in mice and humans. In particular, c-Maf is required for the development and function of mechanosensory neurons terminating in lanceolate endings, Meissner corpuscles and Pacinian corpuscles. c-Maf is a key transcription factor directing the development and function of rapidly-adapting mechanoreceptors and their end organs.

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Project description:Although cells of the immune system experience force and pressure throughout their lifecycle, almost nothing is known about how these mechanical processes regulate the immune response. Immune cells in highly mechanical organs, such as the lung, are constantly exposed to tonic and dynamically changing mechanical cues. Here using reverse genetics, we show that myeloid cells respond to force and alterations in cyclical hydrostatic pressure (CHP) via the mechanosensory ion channel (MSIC) PIEZO1. Unbiased RNA-sequencing from macrophages subjected to CHP reveals a striking state of proinflammatory reprogramming. We report a novel mechanosensory-immune signaling circuit which PIEZO1 initiates in response to CHP, activating c-JUN, upregulating Endothelin-1 (EDN1), and stabilizing HIF1α to facilitate a prolonged program of proinflammatory mediators. Using mice conditionally deficient of PIEZO1 in myeloid cells, and cellular depletion assays, we show infiltrating monocytes respond to cyclical force to recruit neutrophils and clear pulmonary Pseudomonas aeruginosa infection. Furthermore, myeloid PIEZO1 also drove lung pathology in a mouse model of pulmonary fibrosis. Our results demonstrate a novel environmental sensory axis that myeloid cells recognize to mount an inflammatory response, and is the first report showing a physiological role for PIEZO1 and mechanosensation in immunity.

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