Aryal Yam Prasad, Kim Tae-Young, Lee Eui-Seon, An Chang-Hyeon, Kim Ji-Youn, Yamamoto Hitoshi, Lee Sanggyu, Lee Youngkyun, Sohn Wern-Joo, Neupane Sanjiv, Kim Jae-Young
Department of Biochemistry, School of Dentistry, Kyungpook National University, Daegu, South Korea.
Department of Oral and Maxillofacial Radiology, School of Dentistry, Kyungpook National University, Daegu, South Korea.
Front Cell Dev Biol. 2021 Aug 25;9:697243. doi: 10.3389/fcell.2021.697243. eCollection 2021.
miRNAs are conserved short non-coding RNAs that play a role in the modulation of various biological pathways during tissue and organ morphogenesis. In this study, the function of in tooth development, through its loss or gain in function was evaluated. A variety of techniques were utilized to evaluate detailed functional roles of during odontogenesis, including tooth cultivation, renal capsule transplantation, hybridization, real-time PCR, and immunohistochemistry. Two-day tooth cultivation at E13 identified altered cellular events, including cellular proliferation, apoptosis, adhesion, and cytoskeletal arrangement, with the loss and gain of . qPCR analysis revealed alterations in gene expression of tooth-related signaling molecules, including β, , , , , and , when inhibited with and mimic. Also, the inhibition of demonstrated increased mesenchymal localizations of pSMAD1/5/8, alongside decreased expression patterns of and within inner enamel epithelium (IEE) in E13 + 2 days cultivated teeth. Moreover, 1-week renal transplantation of cultivated teeth had smaller tooth size with reduced enamel and dentin matrices, along with increased cellular proliferation and expression along the Hertwig epithelial root sheath (HERS), within the inhibitor group. Similarly, in 3-week renal calcified teeth, the overexpression of did not affect tooth phenotype, while the loss of function resulted in long and slender teeth with short mesiodistal length. This study provides evidence that a suitable level of is required for the modulation of major signaling pathways, including Wnt, Bmp, and , during tooth morphogenesis.
微小RNA（miRNAs）是保守的短链非编码RNA，在组织和器官形态发生过程中参与多种生物途径的调控。在本研究中，通过功能缺失或功能获得来评估其在牙齿发育中的功能。采用了多种技术来评估其在牙发生过程中的详细功能作用，包括牙齿培养、肾被膜移植、杂交、实时定量聚合酶链反应（PCR）和免疫组织化学。在胚胎第13天（E13）进行两天的牙齿培养发现，随着其功能缺失和功能获得，细胞事件发生改变，包括细胞增殖、凋亡、黏附以及细胞骨架排列。定量PCR分析显示，当用抑制剂和模拟物抑制时，牙齿相关信号分子的基因表达发生改变，这些信号分子包括β、、、、、和。此外，在E13 + 2天培养的牙齿中，抑制可使磷酸化SMAD1/5/8的间充质定位增加，同时内釉上皮（IEE）中及的表达模式降低。此外，在抑制剂组中，培养牙齿进行1周肾移植后，牙齿尺寸变小，釉质和牙本质基质减少，同时沿赫特维希上皮根鞘（HERS）的细胞增殖和表达增加。同样，在3周肾移植钙化牙齿中，的过表达不影响牙齿表型，而功能丧失则导致牙齿长而细，近远中长度短。本研究提供了证据，表明在牙齿形态发生过程中，需要适当水平的来调控包括Wnt、骨形态发生蛋白（Bmp）等主要信号通路。
