Fradkin L G, Noordermeer J N, Nusse R
Howard Hughes Medical Institute, Stanford University Medical Center, CA 94035.
Dev Biol. 1995 Mar;168(1):202-13. doi: 10.1006/dbio.1995.1072.
The Wnt gene family encodes highly conserved cysteine-rich proteins which appear to act as secreted developmental signals. Both the mouse Wnt-1 gene and the Drosophila wingless (wg) gene play important roles in central nervous system (CNS) development. wg is also required earlier, in the development of the embryonic metameric body pattern. We have begun to characterize the developmental expression and role of another member of the Drosophila Wnt gene family, DWnt-3. Using antisera raised to the DWnt-3 protein, we show that the protein is secreted in vivo. The early protein expression domains include the limb and appendage primordia. Late expression domains comprise the ventral cord and supraesophageal ganglia of the CNS. Notably, DWnt-3 protein accumulates on the commissural and longitudinal axon tracts of the CNS. Ectopic expression of DWnt-3 in transgenic embryos bearing a HS-DWnt-3 construct leads to specific disruption of the commissural axon tracts of the CNS. We also show that DWnt-3 does not functionally replace wg in an in vivo assay. Experiments with a tissue culture cell line transfected with a construct encoding the DWnt-3 gene show that DWnt-3 protein is efficiently synthesized, glycosylated, proteolytically processed, and transported to the extracellular matrix and medium. DWnt-3, therefore, encodes a secreted protein, which is likely to play a role in development of the Drosophila CNS.
Wnt基因家族编码高度保守的富含半胱氨酸的蛋白质,这些蛋白质似乎作为分泌型发育信号发挥作用。小鼠的Wnt-1基因和果蝇的无翅(wg)基因在中枢神经系统(CNS)发育中都起着重要作用。wg在胚胎分节身体模式的发育中也在更早阶段发挥作用。我们已经开始对果蝇Wnt基因家族的另一个成员DWnt-3的发育表达和作用进行表征。利用针对DWnt-3蛋白产生的抗血清,我们证明该蛋白在体内是分泌型的。早期蛋白表达结构域包括肢体和附属器官原基。晚期表达结构域包括CNS的腹神经索和食管上神经节。值得注意的是,DWnt-3蛋白在CNS的连合轴突束和纵轴突束上积累。在携带HS-DWnt-3构建体的转基因胚胎中异位表达DWnt-3会导致CNS连合轴突束的特异性破坏。我们还表明,在体内试验中DWnt-3不能在功能上替代wg。用转染了编码DWnt-3基因构建体的组织培养细胞系进行的实验表明,DWnt-3蛋白能被高效合成、糖基化、进行蛋白水解加工,并转运到细胞外基质和培养基中。因此,DWnt-3编码一种分泌蛋白,它可能在果蝇CNS的发育中发挥作用。