Functional Analyses of Protocadherin 10 in Embryonic Development of Central Nervous System and Paraxial Mesoderm

研究者和合作者研究了原钙粘蛋白- lo （pcdh10）和其他钙粘蛋白在斑马鱼发育中的生物学功能。原位杂交（ISH）结果显示，pcdh1Ob在松果体、间脑和耳囊附近的近轴中胚层（PAM）和发育中的体中表达。在体细胞成熟过程中，PAM和somite的表达减少。Pcdh1Ob的显性阴性形式和Pcdh1Ob的反义Morpholino干扰了PAM的收敛和somite的分割。对比研究表明，pcdh1Ob和pcdh8在PAM和近轴细胞中的表达存在差异。我们认为Pcdh1Ob参与了PAM的聚合和体小体的分割，Pcdh8和Pcdh10b的粘附差异在体小体和近轴细胞的形态形成机制中起作用。我们使用反义morpholino oligonucletide （MO）联合ISH对斑马鱼肾形成中的钙粘蛋白-6 （cdh6）进行了功能分析。我们克隆了…更多斑马鱼同源的人类k -钙粘蛋白。全负荷ISH显示，cdh6除在中枢和周围神经系统外，还在肾原管和肾原基中表达。注射靶向cdh6的MOs （cdh6MOs）胚胎在30 hpf时出现头小、体轴弯曲、卵黄延伸短、尾短弯曲等发育缺陷，42 hpf时胸部出现水肿。将注射cdh6MOs的胚胎与肾原标记物wtl和pax2.1原位杂交，观察肾原管前区和肾原基发育异常情况。组织学检查显示肾原畸形表现为肾小球原基异常融合，肾原小管融合或异常弯曲，肾原管变薄。这些结果表明，cdh6在斑马鱼胚胎原肾细胞的发育中起着关键作用。我们也研究了斑马鱼的原钙粘蛋白-18。表达于中枢神经系统、咽弓和肠原。目前，研究结果将被汇编成一份科学报告。少

The investigator and collaborators studied biological functions of Protocadherin-lO (Pcdh1O) and other cadherins in zebrafish development. In situ hybridization (ISH) showed that pcdh1Ob was expressed in the paraxial mesoderm (PAM) and developing somites, and in the pineal body, the diencephalon and the vicinity of otocysts. Expression in PAM and somites decreased during somite maturation. A dominant-negative form of Pcdh1Ob and anti-sense Morpholino of pcdh1Ob disturbed convergence of PAM and somite segmentation. Comparative studies showed that differential expressions of pcdh1Ob and pcdh8 in PAM and the adaxial. We propose that Pcdh1Ob is involved in the convergence of PAM and somite segmentation, and that differential adhesion of Pcdh8 and Pcdh10b plays a role in the morphogenic machinery of somites and adaxial cells.We performed functional analyses of cadherin-6 (cdh6) in zebrafish nephrogenesis using antisense morpholino oligonucleotide (MO) inhibition combined with ISH. We cloned … More a zebrafish homolog of human K-cadherin. Whole-mount ISH showed that cdh6 is expressed in the pronephric ducts and nephron primordium in addition to the central and peripheral nervous systems. Embryos injected with MOs directed against cdh6 (cdh6MOs) showed developmental defects including small head, body axis curvature, short yolk extension, and short bent tail by 30 hpf, and edema appeared in the thorax by 42 hpf. Embryos injected with cdh6MOs were subjected to in situ hybridization with pronephric markers, wtl and pax2.1, to examine disturbed development of the anterior region of the pronephric ducts and the nephron primordia. Histological studies showed malformation of the pronephros as abnormally fused glomerulus primordia, fused or abnormally bent pronephric tubule anlage, and thin pronephric ducts. These results suggested that cdh6 plays pivotal roles in development of the pronephros in zebrafish embryos.We also studied zebrafish Protocadherin-18. It was expressed central nervous system, pharyngeal arches and gut primodia. The results are currently to be compiled as a scientific report. Less