Singh Saurabh, Yin Xiaolong, Pisano M Michele, Greene Robert M
Department of Molecular, Cellular and Craniofacial Biology, University of Louisville Birth Defects Center, ULSD, Louisville, Kentucky 40292, USA.
Birth Defects Res A Clin Mol Teratol. 2007 Jan;79(1):35-44. doi: 10.1002/bdra.20320.
Formation of the mammalian orofacial region involves multiple signaling pathways regulating sequential expression of and interaction between molecular signals during embryogenesis. The present study examined the expression patterns of members of the MAPK family in developing murine orofacial tissue.
Total RNA was extracted from developing embryonic orofacial tissue during gestational days (GDs) 12-14 and used to prepare biotinylated cDNA probes, which were then denatured and hybridized to murine MAPK signaling pathways gene arrays.
Expression of a number of genes involved in the (ERK1/2) cascade transiently increased in the embryonic orofacial tissue over the developmental period examined. Numerous members of the SAPK/JNK cascade were constitutively expressed in the tissue. Genes known to play a role in p38 MAPK signaling exhibited constitutive expression during orofacial development. Western blot analysis demonstrated that ERK2/1, p38, and SAPK/JNK kinases are present in embryonic orofacial tissue on each of GD 12, 13, and 14. By using phospho-specific antibodies, active ERK was shown to be temporally regulated during orofacial development. Minimal amounts of active p38 and active SAPK/JNK were detected in orofacial tissue during GDs 12-14.
Our study documents specific expression patterns of genes coding for proteins belonging to the ERK1/2, p38, and SAPK/JNK MAPK families in embryonic orofacial tissue. We also demonstrate that active, phosphorylated forms of ERK1/2 only were detected in the embryonic tissue investigated, suggesting a more central role for members of this family in embryonic orofacial development.
哺乳动物口面部区域的形成涉及多个信号通路,这些信号通路在胚胎发育过程中调节分子信号的顺序表达及相互作用。本研究检测了丝裂原活化蛋白激酶(MAPK)家族成员在发育中的小鼠口面部组织中的表达模式。
在妊娠第12至14天从发育中的胚胎口面部组织中提取总RNA,用于制备生物素化的cDNA探针,然后将其变性并与小鼠MAPK信号通路基因芯片杂交。
在所研究的发育时期,胚胎口面部组织中一些参与细胞外信号调节激酶1/2(ERK1/2)级联反应的基因表达短暂增加。应激激活蛋白激酶/应激活化蛋白激酶(SAPK/JNK)级联反应的众多成员在该组织中持续表达。已知在p38 MAPK信号传导中起作用的基因在口面部发育过程中持续表达。蛋白质免疫印迹分析表明,ERK2/1、p38和SAPK/JNK激酶在妊娠第12、13和14天的胚胎口面部组织中均有表达。通过使用磷酸化特异性抗体,发现活性ERK在口面部发育过程中受到时间调控。在妊娠第12至14天期间,口面部组织中检测到的活性p38和活性SAPK/JNK含量极少。
我们的研究记录了胚胎口面部组织中编码ERK1/2、p38和SAPK/JNK MAPK家族蛋白质的基因的特定表达模式。我们还证明,在所研究的胚胎组织中仅检测到活性磷酸化形式的ERK1/2,这表明该家族成员在胚胎口面部发育中发挥着更核心的作用。