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Proteomics analysis of specific mucosal regions of Salmonella-infected mouse cecum

ABSTRACT: The continuously remodeled extracellular matrix (ECM) plays a pivotal role in gastrointestinal health and disease, yet its precise functions remain elusive. In this project, we employed laser capture microdissection of cecal mucosa combined with low-input proteomics to investigate ECM remodeling during Salmonella-driven inflammation. To complement this, we probed how fibronectin fiber tension is altered using a mechanosensitive peptide probe. While fibronectin fibers in healthy intestinal tissue are typically stretched, fibronectin fiber relaxation occurred exclusively during late-stage infection at 72 hours and was localized to already existing clusters of infiltrated neutrophils in the cecal mucosa of Salmonella-infected mice.

INSTRUMENT(S):

ORGANISM(S): Salmonella Enterica Mus Musculus (mouse)

TISSUE(S): Cecum Mucosa

SUBMITTER: Ronja Rappold

LAB HEAD: Viola Vogel

PROVIDER: PXD056083 | Pride | 2025-05-07

REPOSITORIES: Pride

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			Action	DRS
	DIA_LCM_Ctrl_981B.raw	Raw
	DIA_LCM_Ctrl_982B.raw	Raw
	DIA_LCM_Ctrl_986.raw	Raw
	DIA_LCM_Ctrl_990.raw	Raw
	DIA_LCM_FnBPA5pos_977.raw	Raw

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Publications

Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice.

Rappold Ronja R Kalogeropoulos Konstantinos K La Regina Gianna G Auf dem Keller Ulrich U Slack Emma E Vogel Viola V

npj biological physics and mechanics 20250204 1

The continuously remodeled extracellular matrix (ECM) plays a pivotal role in gastrointestinal health and disease, yet its precise functions remain elusive. In this study, we employed laser capture microdissection combined with low-input proteomics to investigate ECM remodeling during <i>Salmonella</i>-driven inflammation. To complement this, we probed how fibronectin fiber tension is altered using a mechanosensitive peptide probe. While fibronectin fibers in healthy intestinal tissue are typica ...[more]

PMID: 39917413

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Quantitative proteomics analysis of Salmonella-infected mouse cecum

Project description:The ECM is a complex fibrillar network of macromolecules that provides structural and mechanical support to the intestinal tissue. One abundant component of the ECM observed in the Salmonella-driven intestinal edema is the mechanosensitive glycoprotein fibronectin. Combining mechanosensitive staining for relaxed fibronectin with total fibronectin revealed that fibronectin fibers present in the edema are in a tensed molecular conformation. Co-staining with fibrin(ogen) indicates a provisional matrix, similar to what is observed in response to skin injury. The absence of low tensional fibronectin fibers and the additional finding of a high number of protease inhibitors in the edema proteome could indicate a critical role of stretched fibronectin fibers in maintaining tissue integrity in the severely inflamed cecum. Understanding these processes may provide valuable functional diagnostic markers of intestinal disease progression.

2024-09-16 | PXD043256 | Pride

Proteomic analysis of laser-microdissected murine fetal glomeruli

Project description:Alteration of fetal kidney morphogenesis in diabetic pregnancies is poorly described, especially changes in the extracellular matrix (ECM) and glomerular basement membrane (GBM) during glomerulogenesis. Addressing the ECM proteome, or matrisome, in mouse fetal glomeruli in a healthy and diabetic environment will improve understanding about the association between ECM and GBM changes in the glomerulus as potential reprogramming mechanisms for glomerular dysfunction in infants of mothers with diabetes. This project aimed to define the matrisome and BM composition in the maturing murine fetal glomeruli collected from normal and diabetic mothers. For this, we used laser microdissection microscopy to isolate maturing glomeruli from cryosections, and analyse by label-free tandem mass spectrometry to test the hypothesis that maternal diabetes influences ECM and GBM composition, assembly and function.

2021-10-13 | PXD025911 | Pride

Expression profiles of laser dissected colon tumor samples of wild-type mice and vil-Cre-Bcl9-/-/Bcl9l-/- mice

Project description:To investigate the impact of ablating Bcl9/Bcl9l on tumorigenesis, 6-8- week-old mice were exposed first to a single dose dimethylhydrazine (DMH, 44 mg/kg body weight), which is metabolized in the liver to carcinogenic azoxymethane (AOM), followed by 7 days oral administration of 2 % dextrane sulfate sodium (DSS) in the drinking water. This regimen results in the emergence of dysplastic adenomas, which progress to differentiated adenocarcinomas that are morphologically similar to human colorectal adenocarcinomas and typically harbor β-catenin stabilizing mutations of GSK3ß phosphorylation sites. Accordingly, these tumors present hallmarks of active Wnt signaling such as accumulation of nuclear β-catenin and expression of Wnt target genes. Total RNA of laser dissected samples from five different tumors each of two wild-type mice and three vil-Cre-Bcl9-/-/Bcl9l-/- mice was collected and resulting amplified cDNA hybridized to Affymetrix Mouse Genome 430 2.0 arrays. Samples are labeled as follows: Genotype_TumorID_MouseID_UniqueID.

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Testing serum specificity for bacterial antigens in a mouse model of spontaneous colitis

Project description:Identification of bacterial antigens within cecal bacterial lysate was performed by using serum antibodies from control and DC-LMP1/CD40 animals for immunoprecipitation followed by label-free liquid chromatography tandem mass spectrometry (LC-MS/MS). Serum antibodies were coupled to beads and incubated with cecal bacterial lysate to bind target proteins. Upon immunoprecipitation, on-beads digestion of proteins followed by LC-MS/MS was performed. The resulting peak intensities were used for intensity-based quantification (iBAQ). Proteins identified with a fold change > 2 and a p-value < 0.05 were considered for further analyses.

2021-02-25 | PXD018025 | Pride

X-Ray Exposure Induces Structural Changes in Human Breast Proteins

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2025-06-28 | PXD060871 | Pride

Maternal cecal microbiota transfer rescues early-life antibiotic-induced enhancement of type 1 diabetes in mice-the long term restoration

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2021-09-02 | E-MTAB-9982 | biostudies-arrayexpress

The transitional Gene Expression Profiling of Ovarian Follicle Treated with Indomethacin and RU486 in Rats

Project description:Indomethacin (an inhibitor of prostaglandin synthesis) and RU486 (mifepristone, progesterone receptor antagonist) are well known compounds which impair follicle rupture in rats, nevertheless the histopahotlogical figures of these follicle were different in detail due to the difference of the pathway which each compound inhibits. To characterize these unruptured follicles further and to investigate the genes which were altered in each rat treated with indomethacin or mifepristone, gene expression profile in ovarian follicle were analyzed in female sprague-Dawley rats. Peri- or post ovulatory follicles were collected by laser microdissection and extracted RNA was analyzed by genechip after single dosage of mifepristone or indomethacin. Animals were singly administered orally Indomethacin (IM) at 4mg/kg at 16:00 or RU486 (RU) at 100 mg/kg at 10:00 on the proestrus day, and sacrificed by exsanguination under anesthesia at 22:00 on the proestrus day (PeF: peri-ovulatory follicle), and 10:00 on the estrus day (PoF: post-ovulatory follicle). Ovaries were also collected from untreated rats at 22:00 on the proestrus day, and 10:00 on the estrus day as control groups. Three animals were contained per each treatment group at each timing. The ovaries were removed and embedded in OCT (Tissue-Tek, Sakura Finetek, Torrance, CA), frozen in dry-ice-cold isopentan, and stored at -70 C until used. Frozen sections (8 micro meter thick) were mounted on the membrane slides (MMI, Glatteburg, Zurich, Switzerland), and stained with HistogeneTM LCM frozen section staining kit (Arcturus Engineering, Mountain View, CA). Then, 15 follicles in each phase were retrieved by Laser Micro Dissection system (LMD, CellCut Plus, MMI). The collected follicle sections were lysed by the buffer RLT in the collection tube supplied in RNeasy Micro Kit (Qiagen, Hilden, Germany), and total RNA was extracted according to manufacturer's instructions.

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