Ubc9对鸡胚神经嵴细胞发育的影响

The effect of Ubc9 on neural crest cell development of chicken embryo

目的探索Ubc9和SUMO通路对鸡胚神经嵴细胞发育的影响。方法 1)采集鸡胚,通过免疫荧光实验和原位杂交实验来检测SUMO结合酶Ubc9及神经嵴细胞标志基因的表达情况。2)设置对照组(非特异性吗啉代处理)和Ubc9-吗啉代组来处理各组8期鸡胚,随后通过原位杂交实验检测Snail2,Sox9和Fox D3等神经嵴细胞晚期标志基因的表达情况。结果 1)Ubc9与标志基因Pax7共表达于4、6和8期鸡胚神经嵴细胞前体。2)与对照组相比,Ubc9-吗啉代组鸡胚单侧上Snail2(P<0.05),Sox9(P<0.01)和Fox D3(P<0.05)的mRNA水平均有下降。结论Ubc9表达于4、6和8期鸡胚的神经脊细胞前体,鸡胚神经嵴细胞的形成需要SUMO通路参与。

Objective To examine the effect of Ubc9 and SUMO pathway on neural crest cell development of chicken embryo. Methods 1 ）Chicken embryos were collected, In situ hybridization and Immunofluorescence were used to observe the expression of SUMO-conjugating enzyme-Ubc9 and early markers of neural crest. 2）In the control group, 5- mismatch Ubc9-control morpholino （SMM-Mo） was used to treat chick embryos; In the Ubc9-Mo group, sequence-spe- cific Morpholinos （Mo） was used to reduce Ubc9 level in chicken embryos, then the mRNA of neural crest markers （Snail2,Sox9 and FoxD3） was examined. Results 1 ） Ubc9 was coexpressed with Pax7 in neural crest precursors of the neural fold at 4,6 and 8 stage of early chicken embryos. 2）Compared to the control group,Ubc9-Mo treatment led to a reduce of mRNA level of neural crest markers, Snail2（P 〈0. 05）,Sox9 （P 〈0. 01 ）and FoxD3（P 〈0. 05）. Conclusions SUMO-conjugating enzyme, Ubc9 expression start at neural fold at 4,6 and 8 stage of early chicken embryo, Ubc9 and SUMO pathway are required for neural crest cell development of chicken embryo.