大鼠胚胎下頜磨牙發(fā)育初期p75NTR及RUNX2時空表達
發(fā)布時間:2018-04-05 14:03
本文選題:pNTR 切入點:RUNX 出處:《基因組學(xué)與應(yīng)用生物學(xué)》2017年07期
【摘要】:觀察p75神經(jīng)營養(yǎng)受體(p75 neurotrophin receptor,p75NTR)及Runt相關(guān)轉(zhuǎn)錄因子2(runt-related transcription factor 2,RUNX2)在SD大鼠下頜第一磨牙成牙發(fā)育初期不同發(fā)育階段的表達分布情況,并探討兩者在SD大鼠出生后下頜第一磨牙發(fā)育礦化的作用。我們首先制備SD大鼠下頜第一磨牙發(fā)育初期標本(E13.5 d,E14.5 d,E15.5 d,E16.5 d,E18.5 d和P0.5 d),免疫組織化學(xué)檢測p75NTR和RUNX2的表達分布情況。并用表達強度差異較大的天數(shù)P0.5 d的大鼠外胚間充質(zhì)干細胞(ectomesenchymal stem cells,EMSC)進行體外礦化誘導(dǎo),同時本實驗采用蛋白質(zhì)印跡法檢測0 d、7 d、14 d和21 d的p75NTR和RUNX2的表達情況。我們發(fā)現(xiàn)p75NTR在蕾狀期(E13.5 d)開始表達于成釉器旁邊的間質(zhì);帽狀期(E14.5 d,E15.5 d)廣泛表達在內(nèi)釉上皮、釉結(jié)、星網(wǎng)狀層、牙乳頭、牙囊;鐘狀期(E16.5 d,E18.5 d,P0.5 d)表達在內(nèi)釉上皮、釉結(jié)、星網(wǎng)狀層、中間層、牙乳頭,牙囊。RUNX2在蕾狀期、帽狀初期(E13.5 d,E14.5 d)表達在成釉器下方間質(zhì);帽狀末期(E15.5 d)達在在內(nèi)釉上皮、釉結(jié)、星網(wǎng)狀層、牙乳頭、牙囊;鐘狀期(E16.5 d,E18.5 d,P0.5 d)表達在在內(nèi)釉上皮、釉結(jié)、星網(wǎng)狀層、中間層、牙乳頭、牙囊;在帽狀末期到鐘狀期(E15.5 d,E16.5 d,E18.5 d)RUNX2與p75NTR的表達范圍和強度是相似的,但在鐘狀分化期(P0.5 d)RUNX2的表達范圍類似于p75NTR但強度下降。P0.5 d EMSC蛋白質(zhì)印跡法結(jié)果顯示:隨著礦化誘導(dǎo)時間的上升,p75NTR和RUNX2的表達均呈上升趨勢。這些結(jié)果提示我們p75NTR和RUNX2在成牙初期不同天數(shù)表達位置及變化,具有時空特異性,可能與牙齒礦化發(fā)育相關(guān),礦化誘導(dǎo)后變化趨勢趨于一致說明二者在SD大鼠成牙發(fā)育初期下頜第一磨牙的發(fā)育及礦化中都起到一定的作用,可能存在正相關(guān)關(guān)系。
[Abstract]:To observe the expression of p75 neurotrophin receptor p75NTR and 2(runt-related transcription factor 2rUNX2) in different developmental stages of mandibular first molar in SD rats.To explore the role of both of them in the development and mineralization of mandibular first molar after birth in SD rats.First, we prepared the first mandibular molars of SD rats at the early stage of development. The expression of p75NTR and RUNX2 was detected by immunohistochemistry. The specimens of E13.5 d, E14. 5 d, E 15. 5 d, E 16. 5 d and P 0. 5 d were prepared. The expression of p75NTR and RUNX2 were detected by immunohistochemistry.The extracellular mesenchymal stem cells were induced by extracellular mesenchymal stem cells, and the expression of p75NTR and RUNX2 were detected by Western blotting.The stellate reticular layer, middle layer, dental papilla, dental sac. RUNX2 were expressed in the mesenchyma beneath the enamel apparatus at the early stage of cap formation, and in the inner enamel epithelium, enamel knot, stellate reticular layer, dental papilla and dental sac.However, the expression range of P0.5 d)RUNX2 was similar to that of p75NTR, but the intensity decreased. The results of EMSC Western blotting showed that the expression of p75NTR and RUNX2 increased with the increase of mineralization induction time.These results suggest that the expression of p75NTR and RUNX2 in the early days of tooth formation is spatio-temporal specific and may be related to the development of tooth mineralization.The trend of change after mineralization induced showed that both of them played a certain role in the development and mineralization of mandibular first molars in the early stage of tooth development in SD rats, and there may be a positive correlation between them.
【作者單位】: 重慶醫(yī)科大學(xué)附屬第一醫(yī)院口腔科;大坪醫(yī)院野戰(zhàn)外科研究所口腔科;重慶醫(yī)科大學(xué)附屬口腔醫(yī)院;
【基金】:國家自然科學(xué)基金(81271097)資助
【分類號】:R78
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