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FIG. 3. Comparison of Xath5 expression pattern with that of neurogenic and achaete-scute-like genes in stage 40 Xenopus retina. (A) Comparison of X-Notch-1 and Xath5 expression patterns. Double in situ hybridizations were performed as in Fig. 1 with Xath5 in deep purple and X-Notch-1 in red. (A) Xath5 expression is detected in the CMZ except in cells in the most peripheral region. Occasional labeled cells were seen in the central retina, being probably late born cells. (B) X-Notch-1 labeling is also detected in the CMZ except in cells in the most peripheral region. Occasional labeled cells are seen in the central retina. C and D show a higher magnification of the CMZ from the sections depicted in A and B, respectively. (E) Double labeling demonstrates that X-Notch-1 and Xath5 expressions do not exactly overlap, since some X-Notch-1 positive cells do not express Xath5 (arrow). Scale bar in A and B, 100 um; scale bar in C, D, and E, 30um. (F) Comparison of X-Delta-1 and Xath5 expression patterns. Double in situ hybridizations were performed as in Fig. 1 with Xath5 in deep purple and X-Delta-1 in red. (F) Xath5 expression is detected in the CMZ except in cells in the most peripheral region. (G) X-Delta-1 labeling is also detected in the CMZ except in cells in the most peripheral region. H and I show a higher magnification of the CMZ from the sections depicted in F and G, respectively. (J) Double labeling demonstrates that X-Delta-1 and Xath5 expressions do not exactly overlap, since some X-Delta-1 positive cells do not express Xath5 (arrow). Scale bar in F and G, 100 um; scale bar in H, I, and J, 30 um. (K) Comparison of Xath5 and Xash3 expression patterns. Double in situ hybridizations were performed as in Fig. 1 with Xath5 in deep purple and Xash3 in red. (K) Xath5 expression was restricted to the more central part of the CMZ. (L) Xash3 labeling was also restricted to the more central part of the CMZ. M and N show a higher magnification of the CMZ from the sections depicted in K and L, respectively. (O) Double labeling demonstrates that Xash3 expression starts more in the periphery of the CMZ than Xath5 expression and that there is a small area in the CMZ where both expressions overlap (arrow). Scale bar in K and L, 100 um; scale bar in M, N, and O, 30 um. |
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FIG. 4. Comparison of Xath5 and other proneural gene expression patterns in stage 40 Xenopus retina. (A) Comparison of Xath5 and neuroD expression patterns. Double in situ hybridizations were performed as in Fig. 1 with neuroD in deep purple and Xath5 in red. (A) neuroD expression is seen in cells in the CMZ and in the central retina, in the outer part of the inner nuclear layer, and in the outer nuclear layer. (B) Xath5 expression is detected in the CMZ except in cells in the most peripheral region. C and D show a higher magnification of the CMZ from the sections depicted in A and B, respectively. (E) Double labeling demonstrates that Xath5 and neuroD expressions start at the same level in the CMZ. Scale bar in A and B, 100 ﰂm; scale bar in C, D, and E, 30 ﰂm. (F) Comparison of ATH-3 and Xath5 expression patterns. Double in situ hybridizations were performed as in Fig. 1 with ATH-3 in deep purple and Xath5 in red. (F) ATH-3 expression is seen in cells in the CMZ and in the central retina, in the outer part of the inner nuclear layer. (G) Xath5 expression is detected in the CMZ except in cells in the most peripheral region. H and I show a higher magnification of the CMZ from the sections depicted in F and G, respectively. (J) Double labeling demonstrates that Xath5 and ATH-3 expressions start at the same level in the CMZ. Scale bar in F and G, 100 ﰂm; scale bar in H, I, and J, 30 ﰂm. (K) Comparison of X-MyT1 and Xath5 expression patterns. Double in situ hybridizations were performed as in Fig. 1 with this time X-MyT1 in deep purple and Xath5 in red. (K) X-MyT1 expression is seen in cells in the CMZ and in the central retina, in the inner nuclear layer. (L) Xath5 expression is detected in the CMZ except in cells in the most peripheral region. In the central retina, similarly to what was observed for X-Notch-1 and X-Delta-1, a few cells, probably late born cells, express Xath5 in this retina which is at a slightly younger stage than the retina shown in Fig. 3. M and N show a higher magnification of the CMZ from the sections depicted in K and L, respectively. (O) Double labeling demonstrates that Xath5 and X-MyT1 expressions start at the same level in the CMZ. Scale bar in K and L, 100 um; scale bar in M, N, and O, 30 um |
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FIG. 7. Double staining for BrdU uptake and gene expression in stage 40 Xenopus CMZ. (A) Double staining for BrdU uptake and Xath5 expression. Xath5 staining is shown in A, BrdU immunostaining in B. Double staining in C shows that BrdUﰃ cells in the peripheral CMZ are Xath5ﰄ. In the central CMZ, BrdUﰃ cells are stained for Xath5. A few cells are BrdUﰄ and stained with Xath5. (D) Double staining for BrdU uptake and neuroD expression. neuroD staining is shown in D, BrdU immunostaining in E. Double staining in F shows that BrdUﰃ cells in the peripheral CMZ are neuroDﰄ. In the central CMZ, BrdUﰃ cells are stained for neuroD. neuroDﰃ cells in the central retina are BrdUﰄ. (G) Double staining for BrdU uptake and Brn-3.0 expression. Brn-3.0 staining is shown in G, BrdU immunostaining in H. Double staining in I shows that all BrdUﰃ cells in the CMZ are Brn-3.0ﰄ and that all Brn-3.0ﰃ cells are BrdUﰄ. (J) Double staining for BrdU uptake and Pax6 expression. Pax6 staining is shown in J, BrdU immunostaining in K. Double staining in L shows that BrdUﰃ cells in the CMZ express a low level of Pax6 and that cells in the central retina expressing a strong level of Pax6 are BrdUﰄ. Peripheral CMZ is on the left. Scalebar in A, 30 ﰂm. |
