Dataset Information

A COMPREHENSIVE AND QUANTIFIABLE INSIGHT INTO THE PROTEOMES OF ARTICULAR CARTILAGE AND SUBCHONDRAL BONE

ABSTRACT: In this study, we used two workflows based on either pressure cycling technology (PCT) or ultrasonication (US) in combination with data independent acquisition (DIA) based mass spectrometry (MS) analysis to quantitatively assess the proteomes of articular cartilage and subchondral bone.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Equus Caballus (horse)

TISSUE(S): Articular Cartilage, Bone, Synovial Tissue

SUBMITTER: Louise Bundgaard

LAB HEAD: Louise Bundgaard

PROVIDER: PXD025882 | Pride | 2022-10-12

REPOSITORIES: Pride

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Publications

Effective protein extraction combined with data independent acquisition analysis reveals a comprehensive and quantifiable insight into the proteomes of articular cartilage and subchondral bone.

Bundgaard L L Åhrman E E Malmström J J Auf dem Keller U U Walters M M Jacobsen S S

Osteoarthritis and cartilage 20210920 1

<h4>Objective</h4>The objectives of this study was to establish a sensitive and reproducible method to map the cartilage and subchondral bone proteomes in quantitative terms, and mine the proteomes for proteins of particular interest in the pathogenesis of osteoarthritis (OA). The horse was used as a model animal.<h4>Design</h4>Protein was extracted from articular cartilage and subchondral bone samples from three horses in triplicate by pressure cycling technology or ultrasonication. Digested pr ...[more]

PMID: 34547431

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Project description:The purpose of this study was to characterize the histologic development of OA in a mouse model where OA is induced by destabilization of the medial meniscus (DMM model) and to identify genes regulated during different stages of the disease, using RNA isolated from the joint M-bM-^@M-^\organM-bM-^@M-^] and analyzed using microarrays.427 genes from the microarrays passed consistency and significance filters. There was an initial up-regulation at 2 and 4 weeks of genes involved in morphogenesis, differentiation, and development, including growth factor and matrix genes, as well as transcription factors including Atf2, Creb3l1, and Erg. Most genes were off or down-regulated at 8 weeks with the most highly down-regulated genes involved in cell division and the cytoskeleton. Gene expression increased at 16 weeks, in particular extracellular matrix genes including Prelp, Col3a1 and fibromodulin.The results support a phasic development of OA with early matrix remodelling and transcriptional activity followed by a more quiescent period that is not maintained. A group of 9 mice was used for collection of RNA at time 0 (before surgery) when the animals were 12 weeks old. For the other time points, 9 DMM and 9 sham controls were sacrificed at 2, 4, 8, and 16 weeks after surgery for RNA isolation. The tissue included tibial plateau and femoral condyle articular cartilage, subchondral bone with any osteophytes, meniscus, and the joint capsule with synovium was used for RNA isolation. The tissue was treated with RNAlaterM-BM-. (Invitrogen) prior to freezing and storage at -800 C. RNA was extracted by homogenization using the Precellys 24 tissue homogenizer (Bertin Technologies purchased from MO BIO) and the amount and quality of the RNA was determined using an Agilent 2100 Bioanalyzer. RNA was pooled prior to microarray analysis such that 3 randomly selected samples from each surgical group and time point were pooled to create each biological replicate. Because 9 mice were used for each experimental group, a total of three biological replicates per group were analyzed using the Affymetrix Mouse Genome 430 2.0 oligonucleotide arrays as described. One replicate pool, which was from week two DMM mice, did not meet the RNA integrity level needed for microarray analysis; thus, this pool was not analyzed further, leaving two pools for the week two DMM mice.

Dataset Information

A COMPREHENSIVE AND QUANTIFIABLE INSIGHT INTO THE PROTEOMES OF ARTICULAR CARTILAGE AND SUBCHONDRAL BONE

Publications

Effective protein extraction combined with data independent acquisition analysis reveals a comprehensive and quantifiable insight into the proteomes of articular cartilage and subchondral bone.

Similar Datasets

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