猪胚胎颞下颌关节髁状突骨化的转录组学和蛋白质组学研究。
Transcriptomic and proteomic studies of condylar ossification of the temporomandibular joint in porcine embryos.
机构信息
Beijing Research Institute of Traumatology and Orthopaedics, Beijing, China.
Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
出版信息
Animal Model Exp Med. 2023 Aug;6(4):294-305. doi: 10.1002/ame2.12326. Epub 2023 May 31.
BACKGROUND
The ossification mechanism of the temporomandibular joint (TMJ) condyle remains unclear in human embryo. The size and structure of TMJ, shape of articular disc and the characteristics of omnivorous chewing in the pig are similar to those of humans. The pig is an ideal animal for studying the mechanism of ossification of the TMJ condyle during the embryonic period.
METHOD
In a previous study by our group, it was found that there was no condylar ossification on embryonic day(E) 45, but the ossification of condyle occurred between E75 and E90. In this study, a total of 12 miniature pig embryos on E45 and E85 were used. Six embryos were used for tissue sections (3 in each group). The remaining six embryos were used for transcriptomic and proteomic studies to find differential genes and proteins. The differentially expressed genes in transcriptome and proteomic analysis were verified by QPCR.
RESULTS
In total, 1592 differential genes comprising 1086 up-regulated genes and 506 down-regulated genes were screened for fold changes of ≥2 to ≤0.5 between E45 and E85. In the total of 4613 proteins detected by proteomic analysis, there were 419 differential proteins including 313 up-regulated proteins and 106 down-regulated proteins screened for fold changes of ≥2 to ≤0.5 between E45 and E85. A total of 36 differential genes differing in both transcriptome and proteome analysis were found. QPCR analysis showed that 14 of 15 selected genes were consistent with transcriptome analysis.
CONCLUSION
Condylar transcriptome and proteomic analysis during the development of TMJ in miniature pigs revealed the regulatory genes/proteins of condylar ossification.
背景
人类胚胎颞下颌关节（TMJ）髁突的成骨机制尚不清楚。TMJ 的大小和结构、关节盘的形状以及猪的杂食性咀嚼特点与人相似。猪是研究胚胎期 TMJ 髁突成骨机制的理想动物。
方法
本课题组前期研究发现，胚胎第 45 天（E45）时髁突尚无骨化，而 E75 至 E90 之间出现了髁突骨化。本研究共选取 E45 和 E85 的 12 只小型猪胚胎，其中 6 只用于组织切片（每组 3 只），另外 6 只用于转录组和蛋白质组学研究，以寻找差异基因和蛋白。通过 QPCR 验证转录组和蛋白质组分析中差异表达基因的验证。
结果
共筛选出 1592 个差异基因，其中上调基因 1086 个，下调基因 506 个，两组间差异倍数（fold change）≥2 且≤0.5。蛋白质组学分析共检测到 4613 种蛋白，其中上调蛋白 313 种，下调蛋白 106 种，两组间差异倍数（fold change）≥2 且≤0.5。在转录组和蛋白质组分析中均有差异的基因共 36 个。QPCR 分析显示，在 15 个选定基因中，有 14 个与转录组分析一致。
结论
小型猪 TMJ 发育过程中髁突的转录组和蛋白质组分析揭示了髁突骨化的调控基因/蛋白。
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