Fax +41 61 306 12 34 E-Mail karger@karger.ch www.karger.com Original Paper Cells Tissues Organs 2008;188:287–298 DOI: 10.1159/000121610 Notch1, Jagged1, and HES5 Are Abundantly Expressed in Osteoarthritis C. Karlsson C. Brantsing S. Egell A. Lindahl Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden analysis of Notch markers in healthy and OA articular carti- lage were performed. Genes regulated by Notch signalling were studied using microarray. Cytokine-induced transcrip- tion of Notch markers was analyzed using real-time PCR and its effect on cellular localization of the intracellular domain of Notch1 (NICD1) was investigated using immunohisto- chemistry, subcellular fractionation, and transfection. The effect of NF B activation on HES5 transcription was studied using the NF B inhibitor pyrrolidine dithiocarbamate. Re- sults: Notch signalling was activated in OA cartilage and Notch1, Jagged1, and HES5 were abundantly expressed com- Key Words Articular cartilage  Osteoarthritis  Notch signalling Abstract Background: Notch signalling controls differentiation and proliferation in various cell types and is associated with sev- eral diseases. We investigated the localization and regula- tion of several Notch markers in human osteoarthritic (OA) cartilage as well as identified genes controlled by Notch sig- nalling. Methods: Immunolocalization and real-time PCR Accepted after revision: November 26, 2007 Published online: March 20, 2008 Camilla Karlsson Department of Clinical Chemistry, Sahlgrenska Hospital University of Gothenburg, Bruna Stråket 16 SE–413 45 Göteborg (Sweden) Tel. +46 31 342 4591, Fax +46 31 412 250, E-Mail camilla.karlsson@medic.gu.se © 2008 S. Karger AG, Basel 1422–6405/08/1883–0287$24.50/0 Accessible online at: www.karger.com/cto Abbreviations used in this paper ACT autologous chondrocyte transplantation NICD1 Notch intracellular domain 1 BMP2 bone morphogenetic protein-2 OA osteoarthritis CALCRL calcitonin receptor-like PDTC pyrrolidine dithiocarbamate DAPT N-[N-(3,5-diflurophenylacetate)-L-alanyl]- PRG4 proteoglycan 4/lubricin (S)-phenylglycine t-butyl ester Prop1 Prophet of Pit1 DLX4 distal-less homeobox 4 RA rheumatoid arthritis FGFR2 fibroblast growth factor receptor 2 RARRES2 retinoic acid receptor responder 2 HES Hairy and Enhancer of Split Su(H) Suppressor of Hairless HOX homeobox TNF- tumour necrosis factor alpha ID4 inhibitor of differentiation/DNA binding 4 TNFRSF10c tumour necrosis factor receptor superfamily, IGF1BP7 insulin-like growth factor binding protein 7 member 10c IL interleukin TNFSF15 tumour necrosis factor (ligand) superfamily, MME membrane metallo-endopeptidase member 15 MMP9 matrix metalloproteinase-9