ABSTRACT
OBJECTIVE: Chondrocyte hypertrophy followed by cartilage destruction is a crucial step for osteoarthritis (OA) development, however, the underlying mechanism remains largely unknown. The objectives of this study are to identify the gene that may cause cartilage hypertrophy and to elucidate its role on OA pathogenesis. DESIGN: Gene expression profiles of cartilages from OA patients and normal subjects were examined by microarray analysis. Expression of deiodinases, enzymes for regulation of triiodothyronine (T3) biosynthesis, in human and rat articular cartilage (AC) were examined by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Rat ACs and chondrocytes were treated with T3 to investigate its role on chondrocyte hypertrophy and inflammatory reaction. Cartilage-specific Type II deiodinase (DIO2) transgenic rats were generated using bacterial artificial chromosome harboring the entire rat Col2a1 and human DIO2 gene. An experimental OA model was created in the animal to examine the role of DIO2 on cartilage degeneration. RESULTS: DIO2 is highly expressed in OA patient AC compared to normal control. In rat AC, DIO2 is specifically expressed among deiodinases and dominantly expressed the same as in brown adipose tissue. T3 induces hypertrophic markers in articular chondrocyte and cartilage explant culture, and enhances the effect of IL-1α on induction of cartilage degrading enzymes. Importantly, cartilage-specific DIO2 transgenic rats are more susceptible to knee joint destabilization and develop severe AC destruction. CONCLUSION: Our findings demonstrate that upregulated expression of DIO2 in OA patient cartilage might be responsible for OA pathogenesis by enhancing the chondrocyte hypertrophy and inflammatory response.
Subject(s)
Cartilage, Articular/metabolism , Chondrocytes/metabolism , Iodide Peroxidase/biosynthesis , Osteoarthritis, Knee/metabolism , Animals , Arthritis, Experimental/metabolism , Cartilage, Articular/drug effects , Case-Control Studies , Chondrocytes/drug effects , Gene Expression Profiling , Humans , Interleukin-1alpha/metabolism , Iodide Peroxidase/drug effects , Iodide Peroxidase/genetics , Rats , Rats, Transgenic , Reverse Transcriptase Polymerase Chain Reaction , Triiodothyronine/pharmacologySubject(s)
Evoked Potentials , Lambert-Eaton Myasthenic Syndrome/immunology , Lambert-Eaton Myasthenic Syndrome/physiopathology , Nerve Tissue Proteins/immunology , Neuromuscular Junction/physiology , Synaptosomes/physiology , Animals , Diaphragm , Disease Models, Animal , Electric Organ/immunology , Electric Organ/physiology , Evoked Potentials/drug effects , In Vitro Techniques , Mice , Nerve Tissue Proteins/administration & dosage , Neuromuscular Junction/drug effects , Phrenic Nerve , Potassium/pharmacology , Rats , Rats, Inbred Lew , Skates, Fish , Synaptic Transmission/drug effects , Synaptic Transmission/physiology , Synaptosomes/immunologyABSTRACT
Alzheimer's amyloid precursor protein (APP) is a transmembrane protein containing three phosphorylation sites in its cytoplasmic domain. In the present study, we isolated cDNA of APP from electric ray electric lobe (elAPP). This APP (elAPP699) consists of 699 amino acids, contains the beta-amyloid domain and has 80.7% similarity with the human APP695 isoform. The cytoplasmic domain, including three phosphorylation sites, was completely conserved. In the nerve terminals of the cholinergic neuron from the electric ray electric organ, we found elAPP699 existed exclusively in the mature form. We found the phosphorylated form of mature elAPP699 in the nerve terminal as well as in cell body. Immature elAPP699 was not subject to phosphorylation. Our findings indicate that, in neurons, the phosphorylation of APP occurs after maturation.
