Dataset Information

Tissue sampling and homogenization with NIRL enables spatially resolved cell layer specific proteomic analysis of the murine intestine

ABSTRACT: For investigating the molecular physiology and pathophysiology in organs the most exact data will be obtained, if not organ specific cell lines are analysed or the whole organ is homogenized, followed by the analysis of its biomolecules, but if the morphological organisation of the organ can be addressed and in the best case the composition of molecules in single cells of the target organ can be analysed. Laser capture microdissection (LCM) is a technique, which is enabling the selection of specific cells of a tissue for further analysis of their molecules. However, LCM is a time-consuming 2-dimensional technique and optimal results are only obtained, if the tissue is fixed e.g. by formalin. Especially for proteome analysis formalin fixation is reducing the number of identifiable proteins, which is an additional drawback. Recently, it was demonstrated that sampling of fresh-frozen (non-fixed) tissue with an infrared-laser is giving higher yields with respect to the absolute protein amount and number of identifiable proteins than conventional mechanical homogenization of tissues. In this study the applicability of the infrared laser tissue sampling for proteome analysis of different cell layers of murine intestine was investigated, using LC-MS/MS-based differential quantitative bottom-up proteomics. By laser ablation eight consecutive layers of the colon tissue were obtained and analyzed. Beside a clear distinguishability of protein profiles between ascending, descending and transversal colon, we identified in the different intestinal cell layers proteins, which are cell-specific as confirmed with data from the human protein atlas. Thus, for the first time the sampling directly from intact fresh-frozen tissue with 3-dimensional resolution is giving access to the different proteomes of the different cell layers of colon tissue.

INSTRUMENT(S): Orbitrap Fusion, Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Colon

SUBMITTER: Hannah Voß

LAB HEAD: Prof. Dr. Hartmut Schlüter

PROVIDER: PXD033584 | Pride | 2022-06-22

REPOSITORIES: Pride

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			Action	DRS
	Deph_Resolution_Layer1_Replicate1.raw	Raw
	Deph_Resolution_Layer1_Replicate2.raw	Raw
	Deph_Resolution_Layer2_Replicate1.raw	Raw
	Deph_Resolution_Layer2_Replicate2.raw	Raw
	Deph_Resolution_Layer3_Replicate1.raw	Raw

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Publications

Efficacy of military antishock trousers in compensatory and decompensatory hemorrhagic hypotension.

Johnson G G Bond R F RF Stack L B LB Class C A CA Hardebeck J R JR

Circulatory shock 19870101 3

The compensatory cardiovascular response to hemorrhage includes a baroreceptor-induced activation of the sympathetic nervous system resulting in an attempt to reestablish MAP through peripheral vasoconstriction. If the hypotension is not reversed this compensatory vasoconstriction will progress to a loss of vascular tone known as vascular decompensation. The primary purpose of the present study was to compare the effectiveness of military antishock trousers (MAST) applied during the compensatory ...[more]

PMID: 3568281

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