国际口腔医学杂志 ›› 2018, Vol. 45 ›› Issue (3): 313-318.doi: 10.7518/gjkq.2018.03.013
丛雯雯, 张岱尊, 肖文林, 薛令法, 许尧祥
Cong Wenwen, Zhang Daizun, Xiao Wenlin, Xue Lingfa, Xu Yaoxiang
摘要： 目的 构建C57BL/6J小鼠腭突体外培养模型,研究血小板衍化生长因子受体α（PDGFR-α）在小鼠腭突体外培养不同时间段的表达变化。方法 应用悬浮培养法分别培养腭突24、36、48 h,体视显微镜及苏木素-伊红（HE）染色观察腭突发育情况。免疫组织化学检测腭突体外培养不同时间段组织中PDGFR-α的表达定位,Western blot检测腭突体外培养不同时间段组织中PDGFR-α的时空表达变化。结果 体视显微镜和HE染色结果均显示,体外培养24 h,双侧腭突向中线方向靠近生长,间隙变窄;36 h,镜下观察到双侧腭突接触,HE切片观察到腭中嵴上皮缝的形成;48 h,镜下观察到双侧腭突融合,HE切片观察到腭中嵴上皮缝消失。PDGFR-α在腭突体外悬浮培养24 h时表达最高,36及48 h后逐渐降低。结论 本实验改进的腭突体外悬浮培养方法模拟了腭突的体内发育环境,为应用体外腭突培养模型研究腭裂复杂的发病机制奠定了基础。笔者采用的体外悬浮培养方法证明腭突发育相关基因PDGFR-α在体外培养腭突的表达变化与其体内表达是一致的。
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| [1] Setó-Salvia N, Stanier P.Genetics of cleft lip and/or cleft palate: association with other common anoma-lies[J]. Eur J Med Genet, 2014, 57(8):381-393. [2] Dai L, Zhu J, Mao M, et al.Time trends in oral clefts in Chinese newborns: data from the Chinese National Birth Defects Monitoring Network[J]. Birth Defects Res A Clin Mol Teratol, 2010, 88(1):41-47. [3] Velázquez-Aragón JA, Alcántara-Ortigoza MA, Estandia-Ortega B, et al.Gene interactions provide evidence for signaling pathways involved in cleft lip/palate in humans[J]. J Dent Res, 2016, 95(11): 1257-1264. [4] Simioni M, Araujo TK, Monlleo IL, et al.Investiga-tion of genetic factors underlying typical orofacial clefts: mutational screening and copy number varia-tion[J]. J Hum Genet, 2015, 60(1):17-25. [5] Soriano P.The PDGF alpha receptor is required for neural crest cell development and for normal patter-ning of the somites[J]. Development, 1997, 124(14): 2691-2700. [6] Eberhart JK, He X, Swartz ME, et al.MicroRNA Mirn140 modulates Pdgf signaling during palato-genesis[J]. Nat Genet, 2008, 40(3):290-298. [7] Stephenson DA, Mercola M, Anderson E, et al.Pla-telet-derived growth factor receptor alpha-subunit gene (Pdgfra) is deleted in the mouse patch (Ph) mu-tation[J]. Proc Natl Acad Sci USA, 1991, 88(1):6-10. [8] 仵素珍, 康鹏, 肖文林, 等. 血小板衍生生长因子受体α基因在C57BL/6J小鼠胚胎腭突融合过程中表达变化的研究[J]. 口腔医学, 2015, 35(12):1008-1012. Wu SZ, Kang P, Xiao WL, et al.The study of PDGF- α gene expression in the C57BL/6J mouse embryo palate shelves[J]. Stomatology, 2015, 35(12):1008-1012. [9] Shiota K, Kosazuma T, Klug S, et al.Development of the fetal mouse palate in suspension organ culture[J]. Acta Anat (Basel), 1990, 137(1):59-64. [10] 张岱尊, 庄翠竹, 肖文林, 等. C57BL/6J小鼠胚胎腭突体外培养模型的建立[J]. 上海口腔医学, 2013, 22(2):132-136. Zhang DZ, Zhuang CZ, Xiao WL, et al.Establish-ment of palatal organ culture model of C57BL/6J mouse embryos [11] Abbott BD, Pratt RM.Human embryonic palatal epithelial differentiation is altered by retinoic acid and epidermal growth factor in organ culture[J]. J Craniofac Genet Dev Biol, 1987, 7(3):241-265. [12] Brunet CL, Sharpe PM, Ferguson MW.The distribu-tion of epidermal growth factor binding sites in the developing mouse palate[J]. Int J Dev Biol, 1993, 37(3):451-458. [13] Tiwari R, Bargmann W, Bose HR Jr.Activation of the TGF-beta/Smad signaling pathway in oncogenic transformation by v-Rel[J]. Virology, 2011, 413(1): 60-71. |
| [1] | 程波 东耀峻. 基质金属蛋白酶及其组织抑制剂在腭发育中的作用[J]. 国际口腔医学杂志, 2004, 31(03): 195-197. |
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