Platelet-activating factor induces pseudopod formation in calcitonin-treated rabbit osteoclasts

Abstract

We demonstrated previously that platelet-activating factor (PAF), a potent inflammatory mediator, acts on osteoclasts to elevate cytosolic [Ca2+] and stimulate resorption. However, it is not clear whether the effects of PAF on resorptive activity are direct or indirect. In the present study, we investigated the effects of PAF on osteoclast motility. Osteoclasts were isolated from the long bones of neonatal rabbits, and cell motility and morphology were monitored using time-lapse video microscopy. Calcitonin, a hormone known to induce retraction of pseudopods and inhibit resorptive activity, was used to render osteoclasts quiescent. Within 10 minutes of calcitonin treatment (100 ng/ml, final), pronounced retraction of pseudopods was observed in 68 of 112 cells tested. When PAF (200 nM, final) was added 10 minutes after calcitonin treatment, pseudopods were evident 1 h later in 15 of 37 calcitonin-responsive cells tested. In contrast, pseudopods were evident in only 4 of 31 calcitonin-responsive cells treated with control solutions (PAF-vehicle or S-PAF, the biologically inactive stereoisomer of PAF). Pseudopod formation was quantified by measuring the planar area of pseudopods with a computer-based video analysis system. When assessed 60 minutes following PAF treatment, the pseudopod area was significantly greater in PAF-treated cells than in control cells. In some calcitonin-treated osteoclasts, PAF induced pseudopod formation when applied focally using an extracellular micropipette, consistent with a direct action of PAF. We conclude that PAF directly induces pseudopod formation in calcitonin-inhibited osteoclasts, a morphologic response indicative of osteoclast activation.

Lysophosphatidic Acid Signals through Multiple Receptors in Osteoclasts to Elevate Cytosolic Calcium Concentration, Evoke Retraction, and Promote Cell Survival

“…On the other hand, PAF causes retraction of pseudopods with respreading observed within 30 min (41). Moreover, PAF promotes respreading of lamellipodia in calcitonin-pretreated osteoclasts and stimulates bone resorption (42)(43)(44). Interestingly, the morphological responses observed in LPA-treated osteoclasts are distinct from the responses elicited by either calcitonin or PAF.…”

Influência da escovação mecânica sobre a infiltração marginal de restaurações de amálgama de prata associadas a agentes resinosos

“…Nossos resultados indicam que a administração de calcitonina, em animais ovariectomizados, imediatamente após a realização de um defeito ósseo, pode exercer um efeito anabólico direta ou indiretamente sobre esse tecido, já que estudos anteriores demonstraram sua ação sobre os osteoclastos 14,18,24 e, em grau menor, sobre os osteoblastos 18 .…”