|
Display additional annotations [+]
|
| |
Fig. 6. Over-expression of lmx1b and its potential binding partners affects nephrin expression. mRNAs of lmx1b and its potential binding partners were injected either alone (panel A) or in combination (panel B), together with lacZ, into one cell of 2-cell embryos. Red Gal staining followed by nephrin in situ hybridisation was carried out at stage 33/34 in order to assess any glomus phenotype. For each embryo, the injected side (indicated by an asterisk, panels a–d) was compared to the uninjected side (panels e–h). Injection of lmx1b mRNA alone did not induce any significant change in nephrin staining (A, compare panel a to e), whereas injection of lim1 or ldb1 alone resulted in the formation of an enlarged and reduced nephrin domain respectively (A, b and f; A, c and g). These phenotypes can be partially rescued by the co-injection of lmx1b (B, b and f; B, c and g). Co-injection of lim1 and ldb1 rescued both phenotypes, the embryos displayed on the injected side a nephrin domain, similar in area to the uninjected side (B, a and e). Injection of E47 alone or in combination with lmx1b did not induce any statistically significant phenotype (A, d and h and B, d and h). |
|
Display additional annotations [+]
|
| |
Fig. 5. Over-expression of lmx1b and its potential binding partners affects wt1 expression. mRNAs of lmx1b and its potential binding partners were injected either alone (panel A) or in combination (panel B), together with lacZ, into one V2 blastomere at the 8-cell stage. Red Gal staining followed by wt1 in situ hybridisation was carried out at stage 35/36 in order to assess any glomus phenotype. For each embryo, the injected side (indicated by an asterisk, panels a–d) was compared to the uninjected side (panels e–h). Injection of lmx1b mRNA alone did not induce any significant glomus phenotype (A, compare panel a to e), whereas injection of lim1 or ldb1 alone, its potential binding partners, resulted in the formation of an enlarged and reduced glomus respectively (A, b and f; A, c and g). These phenotypes can be partially rescued by the co-injection of lmx1b (B, b, arrowhead and f; B, c and g). Interestingly, co-injection of lim1 and ldb1 rescued both phenotypes, resulting in the formation of a more normal glomus (B, a and e). Injection of E47 alone or in combination with lmx1b did not induce any statistically significant phenotype (A, d and h and B, d and h). |
|
Display additional annotations [+]
|
| |
Fig. 7. Over-expression of lmx1b and its potential binding partners affects the development of tubules. mRNAs of lmx1b and its potential binding partners were injected either alone (panel A) or in combination (panel B), together with lacZ into one V2 blastomere at the 8-cell stage. The morphology of the tubules was assessed at stage 40 by immunohistochemistry using 3G8 (in purple) and 4A6 (in red) monoclonal antibodies (Vize et al., 1995). The injected side was identified by blue β-galactosidase staining and is indicated by an asterisk (a–d). As comparison, the uninjected side of each embryo was photographed (e–h) lmx1b-injected embryos showed no pronephric phenotype (A–a and e). lim1 injection resulted in the formation of an enlarged proximal tubule mass and wider more distal tubules (A–b and f), whereas ldb1 over-expression caused reduction in size of proximal tubules and in some cases affected formation of the more distal tubules (A–c and g). Co-injection of lmx1b with either lim1 or ldb1 partially rescued these phenotypes (B–b and f and B–c and g). Co-expression of lim1 and ldb1 partially rescued both lim1 and ldb1 phenotypes (B–a and e). Injection of E47 resulted in the formation of slightly enlarged proximal tubule mass without affecting the more distal tubules (A–d and h) whereas co-injection of lmx1b and E47 caused the opposite effect with reduction of pronephric proximal tubules (B–d and h). |
