Ulatory element, resulting in cell-type-specific altered expression of the gene136. The Wsh mutation arises spontaneously from crossing two inbred strains of C3H/HeH and 101/H mice. The function of KL/c-Kit signaling within the regulation of bone metabolism has been studied in vitro and in vivo. The human osteoblast-like cell line Saos-2 expresses KL on its cell surface, whereas the osteoclast progenitor-like cell line FLG 29.1 expresses c-Kit7. Depending on these research, it was concluded that the c-Kit receptor mediates cell-to-cell interactions amongst osteoclasts and osteoblasts/stromal cells via membrane-bound KL. Previous studies have identified skeletal alterations in Sl/Sld mutants lacking the transmembrane type of KL but had normal c-Kit receptors17. Deletion of membrane-bound KL induces osteopenia. The unfavorable bone balance observed in these mice was mainly on account of elevated osteoclast surface. It has been shown that 14-week-old female WBB6F1/J-KitW/W-v (W/Wv) mice carrying a compound c-Kit mutation have been osteopenic18. Nonetheless, these miceReceived: 20 April 2016 Accepted: 21 July 2016 Published: 16 AugustDepartment of SSTR5 Gene ID Physiology and STAR on Craniofacial and Skeletal Disorders, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand. 2Department of Oral Medicine, Infection, and Immunity, Harvard College of Dental Medicine, Boston, MA, USA. AP-1 web 3Department of Physiology, Faculty of Science, Mahidol University, Bangkok, Thailand. Correspondence and requests for components needs to be addressed to S.L. (e mail: [email protected])Scientific RepoRts six:31515 DOI: 10.1038/srepwww.nature.com/scientificreports/were infertile resulting from a lack of germ cells inside the ovary and had decreased estrogen and progesterone levels, top to elevated FSH level19. It is unclear no matter whether the observed skeletal phenotype in W/Wv mice resulted from cell-autonomous effects in osteoclasts or was a consequence of changes in sex hormone levels. In the present study, we focused around the skeletal phenotype of C57BL/6J-KitW-sh/W-sh (Wsh/Wsh) mice that were totally fertile and determined the mechanism by which altered c-Kit signaling affected bone turnover. Our data indicated that the c-Kit Wsh mutation resulted in decreased cancellous bone volume with a rise in bone resorption and bone formation in developing mice. Calvarial osteoblasts derived from Wsh/Wsh mice showed a rise in osteoblast precursors, alkaline phosphatase (ALP) activity and mineralization in vitro. Furthermore, the RANKL/OPG ratio was enhanced in osteoblasts derived from these mice, major to increased quantity of osteoclasts in c-Kit mutants. Quantitative real-time PCR (qPCR) indicated that c-Kit expression was decreased in Wsh/Wsh bone marrow macrophage (BMM) and osteoclasts but not osteoblasts, suggesting that increased bone formation in Wsh/Wsh mice was not an osteoblasts intrinsic effect. Conditioned medium derived from Wsh/Wsh osteoclasts contained increased levels in the osteoclast-derived coupling aspect Wnt10b, and enhanced ALP activity and mineralization by osteoblasts. Blocking Wnt10b activity inhibited these effects. These findings demonstrate that c-Kit regulates bone turnover by suppressing osteoblast and osteoclast differentiation. Hence, c-Kit mutation increased bone formation by increasing the generation of osteoclast-derived coupling aspect Wnt10b.W/Wv mice exhibited osteopenic phenotype. Earlier studies indicated that each male and female W/Wv mice are infertile19,20. To figure out whet.
