In situ hybridization study of cytokeratin 4, 13, 16 and 19 mRNAs in human developing junctional epithelium
In situ hybridization study of cytokeratin 4, 13, 16 and 19 mRNAs in human developing junctional epithelium
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DOI：
10.1111/j.1600-0722.1997.tb00224.x
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发表时间：
1997-12-01
影响因子：
1.9
通讯作者：
Ouhayoun, JP
中科院分区：
文献类型：
--
作者：
Feghali-Assaly, M;Sawaf, MH;Ouhayoun, JP
细胞角蛋白（CKs）被认为是跟踪上皮组织发育和分化的可靠标志物。应用单克隆抗体和显微组织双向凝胶电泳技术，对人部分萌出牙结合上皮细胞（JE）中的CK 19、CK 16、CK 14、CK 13、CK 6、CK 5、CK 4进行了鉴定。CK谱与发育中的口腔上皮相似，提示萌出期牙齿中的结合上皮是牙源性的。本研究采用原位杂交技术检测了CKs 19、16、13和4的mRNA在人发育中的结合上皮中的分布，并探讨了mRNA与其编码蛋白之间的相关性。CK 19 mRNA在初级结合上皮（PJE）基底层细胞中含量丰富，而在基底上层细胞中含量较少。CK16、13和4mRNA在PJE的基底细胞层中含量丰富。副基底层细胞层强烈反应的cRNA探针互补的CK16 mRNA，是在PJE的基底层上的细胞层的CK 13和4探针的反应。我们的研究结果表明，PJE表达的基因编码远CKs 16和4已揭示以前只通过电泳。因此，他们证实PJE是一种分化良好的复层上皮，具有复杂的独特表型，可产生基底细胞特异性CK（CK 19）、与过度增殖相关的CK（CK 16）以及最终与分层相关的CK（CK 4和13）。只有CK19蛋白和mRNA的合成是严格平行的. CK 4和CK 13 mRNA在基底细胞和基底上细胞中均有表达，而CK 13在基底上细胞层中表达量与CK 13蛋白表达量一致。
Cytokeratins (CKs) are now considered to be reliable markers for following the development and differentiation of epithelial tissue. We have investigated the pathway of differentiation in human developing junctional epithelium using monoclonal antibodies and two-dimensional gel electrophoresis of microdissected tissue to identify CK 19, CK 16, CK 14, CK 13, CK 6, CK 5, CK 4 in the junctional epithelium (JE) over partially erupted human teeth. The CK profile was similar to that of developing oral epithelia, suggesting that the junctional epithelium in teeth during eruption is of odontogenic origin. The present study used in situ hybridization to determine the distribution of the mRNAs of CKs 19, 16, 13 and 4 in human developing junctional epithelium and to examine the correlation between mRNAs and their encoded proteins. CK 19 mRNA was abundant in the basal cell layers of the primary junctional epithelium (PJE) but less concentrated in the suprabasal layers. CK16, 13 and 4 mRNAs were abundant in the basal cell layers of the PJE. The parabasal cell layers reacted intensely to the cRNA probe complementary to CK16 mRNA, as were the reactions in the suprabasal cell layers of the PJE for the CK 13 and 4 probes. Our results demonstrate that the PJE express the genes encoding far CKs 16 and 4 that have been revealed previously only by electrophoresis. They therefore confirm that the PJE is a well-differentiated stratified epithelium with a complex unique phenotype that produces CKs specific for basal cells (CK 19), CKs associated with hyperproliferation (CK 16), and finally those associated with stratification (CKs 4 and 13). Only synthesis of CK 19 protein and mRNA are strictly parallel. CKs 4 and 13 mRNAs are present in basal and suprasal cells, while their encoded proteins were not, except for CK 13 in suprabasal cell layers of PJE, where the amount of its mRNAs was coincident with the expression of the protein.