Cadherin-6 is required for zebrafish nephrogenesis during early development
Cadherin-6 is required for zebrafish nephrogenesis during early development
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DOI：
10.1387/ijdb.062200fk
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发表时间：
2007-01-01
影响因子：
0.7
通讯作者：
Yorifuji, Hiroshi
中科院分区：
文献类型：
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作者：
Kubota, Fumitaka;Murakami, Tohru;Yorifuji, Hiroshi
我们利用反义Morpholino寡核苷酸（MO）抑制结合原位杂交技术对斑马鱼肾形成中的钙粘蛋白-6 （cdh6）进行了功能分析。我们克隆了一个人K-cadherin （CDH6）的斑马鱼同源基因（登录号AB193290），其同源性为60-63%，与人、小鼠、鸡和爪蟾的同源性为76-78%。原位杂交结果显示，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。在受精后14小时（hpf）首次检测到肾原管中cdh6的表达，并增加到24 hpf。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、尾弯短等发育缺陷，42 hpf时胸部出现水肿。这些缺陷和水肿在52 hpf时更为明显，大多数受影响的胚胎在受精后5天死亡。注射cdh6MOs的胚胎用肾原标记物wt1和pax2.1探针原位杂交，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，肾原小管原素融合或异常弯曲，肾原管缩窄。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。
We performed functional analyses of cadherin-6 (cdh6) in zebrafish nephrogenesis using antisense Morpholino oligonucleotide (MO) inhibition combined with in situ hybridization. We have cloned a zebraf ish homolog (accession number AB193290) of human K-cadherin (CDH6), which showed 60-63% identity and 76-78% similarity to the human, mouse, chicken and Xenopus homologs. Whole-mount in situ hybridization showed that cdh6 is expressed in the pronephric ducts and nephron primordia in addition to the central and peripheral nervous systems. Expression of cdh6 in the pronephric ducts was first detected at 14 hours post-fertilization (hpf) and increased to 24 hpf. Embryos injected with MOs directed against cdh6 (cdh6MOs) showed developmental defects, including a small head, body axis curvature, short yolk extension and a short bent tail by 30 hpf and edema appeared in the thorax by 42 hpf. Such defects and edema became more marked by 52 hpf and most of the affected embryos died by 5 days post-fertilization. Embryos injected with cdh6MOs were subjected to in situ hybridization with probes for the pronephric markers, wt1 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 anlagen and coarctated pronephric ducts. These results suggest that cdh6 plays pivotal roles in the development of the pronephros in zebrafish embryos.