Thomas T, Voss A K, Gruss P
Department of Molecular Cell Biology, Max Planck Institute of Biophysical Chemistry, Göttingen, Germany.
Dev Dyn. 1998 Jun;212(2):250-7. doi: 10.1002/(SICI)1097-0177(199806)212:2<250::AID-AJA9>3.0.CO;2-G.
We have generated a gene trap insertion into the protein tyrosine phosphatase-BL (PTP-BL) locus, which produces a fusion of the N-terminal half of PTP-BL with beta-galactosidase. During development, beta-galactosidase activity was seen in all epithelial cells: strong staining was observed in the stomach and kidney epithelium, the ependymal layer of the central nervous system, and the surface ectoderm. Particularly prominent beta-galactosidase activity was seen in the peripheral nervous system, which correlated with neurite outgrowth. In epithelial cells, staining was seen in the apical portion of the cells. In nerves, beta-galactosidase activity was associated with growth cones as well as with Schwann cells. This suggests that the amino-terminal portion of PTP-BL contains sequences sufficient to target the fusion protein to specific subcellular compartments. In situ hybridization with a PTP-BL probe demonstrated that all tissues in which beta-galactosidase activity was seen were genuine sites of expression of the PTP-BL gene, although differences in the stability of the PTP-BL protein and the PTP-BL-beta-galactosidase fusion protein may exist. The distribution of beta-galactosidase activity in the peripheral nervous system, together with the structure of the wild-type protein, suggests that this phosphatase may have a role in regulation of the cytoskeleton during the development of the peripheral nervous system.
我们已在蛋白酪氨酸磷酸酶-BL(PTP-BL)基因座中产生了一个基因陷阱插入,该插入导致PTP-BL的N端一半与β-半乳糖苷酶融合。在发育过程中,在所有上皮细胞中均可见β-半乳糖苷酶活性:在胃和肾上皮、中枢神经系统的室管膜层以及表面外胚层中观察到强染色。在周围神经系统中可见特别突出的β-半乳糖苷酶活性,这与神经突生长相关。在上皮细胞中,染色见于细胞的顶端部分。在神经中,β-半乳糖苷酶活性与生长锥以及施万细胞相关。这表明PTP-BL的氨基末端部分包含足以将融合蛋白靶向特定亚细胞区室的序列。用PTP-BL探针进行原位杂交表明,所有观察到β-半乳糖苷酶活性的组织都是PTP-BL基因的真正表达位点,尽管PTP-BL蛋白和PTP-BL-β-半乳糖苷酶融合蛋白的稳定性可能存在差异。β-半乳糖苷酶活性在周围神经系统中的分布,连同野生型蛋白的结构,表明这种磷酸酶可能在周围神经系统发育过程中对细胞骨架的调节中起作用。