Heat-shock induction time and levels of loss of bone mineralization in the osteoporosis medaka fish model rankl:HSE:CFP subline c1c8
Main Article Content
Abstract
The transgenic medaka fish (Oryzias latipes) rankl:HSE:CFP expressing Rankl, a stimulator for osteoclastogenesis, is used as an osteoporosis model to screen for antiosteoporosis substances. In this fish, rankl expression is regulated by a heat inducible promotor. Therefore, upon heatshock at 39oC, the fish expresses ectopic Rankl, which promotes formation and activity of osteoclasts, leading to bone mineralization damage and osteoporosis-like phenotype. However, how the level of mineralization damage depends on heatshock induction time still has been an open question that we aimed to answer in this study. To this end, 9-day-post-fertilization (dpf) rankl:HSE:CFP fish larvae of subline c1c8 were divided into 5 groups that were heatshocked at 39°C for 30, 60, 75, 90 or 120 minutes. Level of loss of bone mineralization of fish larvae in each group was assessed at 11dpf by their bone mineralization index. Results showed that levels of bone loss of these fish groups were 33, 62, 65, 79 or 93%, respectively, and correlated well and directly proportional with heat-shock duration. These results provide important data that help establishing procedures and protocols for using the fish in further studies on bone.
References
2. Phạm Văn Cường, Phạm Thị Thanh, Nguyễn Thúy Hoa, Trần Đức Long, Tô Thanh Thúy (2015), “Tách dòng cá medaka chuyển gen rankl:HSE:CFP dùng làm mô hình nghiên cứu bệnh loãng xương”, Tạp chí Khoa học ĐHQGHN: Khoa học tự nhiên và Công nghệ, 31 (4S), pp. 24-34.
3. Phạm Thị Thanh , Phạm Văn Cường, Trần Đức Long, Tô Thanh Thúy (2016), “Sự phát triển xương của cá Medaka (Oryzias latipes) II”, Tạp chí Sinh lý học Việt Nam, 20(2), pp. 22-29.
4. Bajoghli B., N. Aghaallaei, T. Heimbucher and T. Czerny (2004), “An artificial promoter construct for heat-inducible misexpression during fish embryogenesis”, Dev Biol, 271 (2), pp. 416-430.
5. Baofeng L., Y. Zhi, C. Bei, M. Guolin, Y. Qingshui and L. Jian (2010), “Characterization of a rabbit osteoporosis model induced by ovariectomy and glucocorticoid”, Acta Orthop, 81 (3), pp. 396-401.
6. Grabher C.,J. Wittbrodt (2008), “Recent advances in meganuclease-and transposon-mediated transgenesis of medaka and zebrafish”, Methods Mol Biol, 461, pp. 521-539.
7. Halade G.V., M.M. Rahman, P.J. Williams and G. Fernandes (2010), “High fat diet-induced animal model of age-associated obesity and osteoporosis”, J Nutr Biochem, 21 (12), pp. 1162-1169.
8. Hwang W.Y., Y. Fu, D. Reyon, M.L. Maeder, S.Q. Tsai, J.D. Sander, R.T. Peterson, J.R. Yeh and J.K. Joung (2013), “Efficient genome editing in zebrafish using a CRISPR-Cas system”, Nat Biotechnol, 31 (3), pp. 227-229.
9. Komori T. (2015), “Animal models for osteoporosis”, Eur J Pharmacol, 759, pp. 287-294.
10. Lin C.Y., C.Y. Chiang and H.J. Tsai (2016), “Zebrafish and Medaka: new model organisms for modern biomedical research”, J Biomed Sci, 23, pp. 19.
11. Naruse K., Tanaka M., and Takeda H. (2011), Medaka - A Model for Organogenesis Human Disease and Evolution, Springer, Japan.
12. Renn J., A. Buttner, T.T. To, S.J. Chan and C. Winkler (2013), “A col10a1:nlGFP transgenic line displays putative osteoblast precursors at the medaka notochordal sheath prior to mineralization”, Dev Biol, 381 (1), pp. 134-143.
13. Seruggia D.,L. Montoliu (2014), “The new CRISPR-Cas system: RNA-guided genome engineering to efficiently produce any desired genetic alteration in animals”, Transgenic Res, 23 (5), pp. 707-716.
14. Shanthanagouda A.H., B.S. Guo, R.R. Ye, L. Chao, M.W. Chiang, G. Singaram, N.K. Cheung, G. Zhang and D.W. Au (2014), “Japanese medaka: a non-mammalian vertebrate model for studying sex and age-related bone metabolism in vivo”, PLoS One, 9 (2), pp. e88165.
15. Thermes V., C. Grabher, F. Ristoratore, F. Bourrat, A. Choulika, J. Wittbrodt and J.S. Joly (2002), “I-SceI meganuclease mediates highly efficient transgenesis in fish”, Mech Dev, 118 (1-2), pp. 91-98.
16. To T.T., P.E. Witten, A. Huysseune and C. Winkler (2015), “An adult osteopetrosis model in medaka reveals the importance of osteoclast function for bone remodeling in teleost fish”, Comp Biochem Physiol C Toxicol Pharmacol, 178, pp. 68-75.
17. To T.T., P.E. Witten, J. Renn, D. Bhattacharya, A. Huysseune and C. Winkler (2012), “Rankl-induced osteoclastogenesis leads to loss of mineralization in a medaka osteoporosis model”, Development, 139 (1), pp. 141-150.
18. Watson A.T., A. Planchart, C.J. Mattingly, C. Winkler, D.M. Reif and S.W. Kullman (2017), “From the Cover: Embryonic Exposure to TCDD Impacts Osteogenesis of the Axial Skeleton in Japanese medaka, Oryzias latipes”, Toxicol Sci, 155 (2), pp. 485-496.