Interaction of cytokines and cell adhesion molecules during osteoclastogenesis

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Most type of cells need to contact with extracellular matrix, which are mainly mediated with integrins. The signal transduction pathway through integrin shares many parts with that of cyokines. The expression of integrin avβ5 is abundant on osteoclast precursors, which need many kinds of cyokines, including M-CSF, for their maturation and survival. Therefore we explored interaction of integrin and cytokine signal pathway on osteoclast precursors.Osteoclast precursors were isolated from C3H mouse hindlimb bones and cultured in PFA dishes for 3 days with M-CSF. Osteoclast precursors were collected and cultured on plastic(A) or PFD(NA) dishes with(+) or without(-) M-CSF. Cell lysate from each group were fractionated on SDS-PAGE, blotted onto membrane and proved by anti-phosphorlated tyrosine.The 50kDa band were regulated by cell attachment and decreased in NA compared with A. The effect of M-CSF was not clear between NA- and NA+, the intensity of 50kDa band in A+ showed 1.3fold increase of that in A-.The 50kDa protein is the candidate of signal transduction molecules. Cell adhesion is necessary for the effect of M-CSF since the intensity of 50kDa band was up-regulated by M-CSF in A.