Table 1. Quantification of Müller glia and specific neuronal classes during retinal regeneration.
Dark-adapted adult albino zebrafish were either uninjected or injected and electroporated with: the GeneTools Standard Control morpholino; a 5-base mismatch control morpholino (5mis pax6a MO and 5mis pax6b MO), anti-pax6a morpholino, anti-pax6b morpholino, or a cocktail of both anti-pax6 morpholinos. Expression of Müller glia and neuronal-specific cell markers were assessed after various times of constant light treatment by immunohistochemistry. Quantification of cell classes was performed on dorsal retinal sections within a 740 micron linear distance equidistant from the margin and the optic nerve. Number of retinas examined (N) and the significance level of the ANOVA analysis (p) are shown for each experiment. Standard deviations are shown in parentheses and asterisks represent data is significantly different from the controls using Bonferroni-corrected t-tests.
| Average percentage of glutamine synthetase-positive Müller glia that are PCNA-positive | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 36 hrs light txt | 6 | 0.792 | 54.6 (2.8) | 48.1 (5.8) | N.A. | N.A. | 51.3 (2.5) | 53.2 (4.5) | 49.8 (3.7) |
| Average number of PCNA-positive nuclei per Müller glial cluster | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 72 hrs light txt | 6 | <0.001 | 7.9 (0.7) | 7.3 (0.5) | 7.7 (0.1) | 8.1 (0.3) | 3.8 (0.1) * | 1.9 (0.2) * | 2.1 (0.2) * |
| Average number of BrdU-positive nuclei per Müller glial cluster | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 72 hrs light txt | 10 | <0.001 | 7.4 (0.6) | N.A. | 8.0 (0.1) | 8.1 (0.1) | 4.6 (0.1) * | 2.3 (0.1) * | 2.0 (0.1) * |
| Average number of PCNA-positive nuclei in ONL | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 3 days post txt | 6 | <0.001 | 76.2 (4.5) | 75.8 (5.6) | 78.4 (5.0) | 70.5 (5.6) | 118.4 (20.3) | 161.0 (11.4) * | 158.6 (11.7) * |
| Average number of Müller glial cells | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 28 days post txt | 8 | 0.827 | 70.6 (2.6) | 69.1 (4.0) | N.A. | N.A. | 71.4 (1.1) | 68.2 (2.7) | 69.0 (1.8) |
| Average number of ONL rod photoreceptor nuclei | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 28 days post txt | 8 | 0.845 | 315.8 (12.0) | 307.6 (12.4) | 335.4 (12.7) | 308.5 (11.0) | 318.4 (16.3) | 322.2 (17.5) | 313.6 (16.4) |
| Average number of long single cone cells | |||||||||
| Time point | N | p | Uninjected | Std. Ctl. MO | 5mis pax6a MO | 5mis pax6b MO | pax6a MO | pax6b MO | pax6a/6b MO |
| 28 days post txt | 8 | <0.001 | 49.2 (5.1) | 48.2 (4.5) | 50.2 (4.8) | 48.4 (5.5) | 34.8 (2.3) | 20.8 (2.8) * | 19.0 (1.9) * |