Comparative study of protein and mRNA expression during osteoclastogenesis.
Osteoclasts, the bone-digesting cells, are key players in bone remodeling. To identify proteins potentially involved in osteoclast function, we analyzed the patterns of protein expression during osteoclastogenesis by2-D DIGE. As a model system we used the mouse myeloid Raw 264.7 cell line that differentiates in vitro into osteoclasts upon treatment with specific growth factors. In 2-D DIGE, we identified 86 up- and 34 down-regulated proteins including known osteoclast differentiation markers as well as proteins regulating key cellular functions of osteoclasts such as energy production, cytoskeleton dynamics, and digestion of organic and inorganic bone matrix. Comparison of protein expression using 2-D DIGE techniques with mRNA expression analyzed by DNA microarrays revealed essentially two groups of genes. The first group comprises genes for which differences in both mRNA and protein expressions were found. A second group covers genes whose expression was not altered at the mRNA level but whose corresponding gene products exhibited different electrophoretic mobilities, thereby revealing potential changes in post-transcriptional processing and PTM. Thus, these combined approaches identify new potential therapeutic targets for treatment of bone diseases and provide complementary information on regulatory processes that might affect osteoclastogenesis.
Czupalla
Cornelia
C
Technical University of Dresden, Dresden, Germany.
Mansukoski
Hannu
H
Pursche
Theresia
T
Krause
Eberhard
E
Hoflack
Bernard
B
D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D008562:Membrane Glycoproteins / Q000494:pharmacology; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D053245:RANK Ligand; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism*; D053246:Receptor Activator of Nuclear Factor-kappa B
D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D011506:Proteins; D053245:RANK Ligand; D012333:RNA, Messenger; D053246:Receptor Activator of Nuclear Factor-kappa B; C505687:Tnfrsf11a protein, mouse; C505689:Tnfsf11 protein, mouse