Antisense inhibition of AMEL translation demonstrates supramolecular controls for enamel HAP crystal growth during embryonic mouse molar development.
Antisense inhibition of AMEL translation demonstrates supramolecular controls for enamel HAP crystal growth during embryonic mouse molar development.
复制标题
DOI：
--
复制
发表时间：
1993-02
期刊：
影响因子：
4.6
通讯作者：
Thomas GH Diekwisch;Sasson David;P. Bringas;V. Santos;Harold C. Slavkin
中科院分区：
文献类型：
--
作者：
Thomas GH Diekwisch;Sasson David;P. Bringas;V. Santos;Harold C. Slavkin
在牙齿发育过程中，牙釉质器官上皮细胞表达组织特异性基因产物（釉原蛋白），其可能在牙釉质生物矿化过程中起到控制羟基磷灰石钙晶体生长模式的作用。本研究旨在检验牙釉蛋白作为超分子聚集体在牙釉质生物矿化过程中调节晶体生长的假设。在简单的器官培养系统中使用反义寡脱氧核苷酸策略来抑制牙釉蛋白翻译。在这些实验条件下，在牙釉蛋白表达之前和期间的反义处理导致免疫沉淀的[35S]甲硫氨酸代谢标记蛋白内的牙釉蛋白翻译产物受到抑制。为了确定该模型系统中反义治疗的效率，观察到地高辛标记的寡脱氧核苷酸在 24 小时内扩散到整个牙齿外植体，包括目标成釉细胞。对反义处理培养物中作为电子致密点画材料的牙釉蛋白超分子组装体的超微结构分析表明，细胞外牙釉质基质的形态异常，晶体长度和宽度显着减少。我们得出的结论是，分泌的细胞外蛋白形成超分子聚集体，它控制牙齿发育过程中牙釉质晶体形成的方向和尺寸。
During tooth development, enamel organ epithelial cells express a tissue-specific gene product (amelogenin) which presumably functions to control calcium hydroxyapatite crystal growth patterns during enamel biomineralization. The present studies were designed to test the hypothesis that amelogenin as a supramolecular aggregate regulates crystal growth during enamel biomineralization. Antisense oligodeoxynucleotide strategy was used in a simple organ culture system to inhibit amelogenin translation. Under these experimental conditions, antisense treatment prior to and during amelogenin expression resulted in inhibition of amelogenin translation products within immunoprecipitated [35S]methionine metabolically labeled proteins. To determine the efficiency of antisense treatment in this model system, digoxigenin-labeled oligodeoxynucleotides were observed to diffuse throughout the tooth explants including the target ameloblast cells within 24 hours. Ultrastructural analyses of amelogenin supramolecular assembly as electron-dense stippled materials in antisense treated cultures demonstrated dysmorphology of the extracellular enamel matrix with a significant reduction in crystal length and width. We conclude that secreted extracellular proteins form a supramolecular aggregate, which controls both the orientation and dimensions of enamel crystal formation during tooth development.