Percentage colocalization of melanopsin and retrogradely transported tracer in RGCs that project to various retinorecipient brain regions
| Injection site per animal | Sections counted | Double-labeled RGCs (raw count) | Double-labeled RGCs per section (corrected) | Percentage of tracer-positive RGCs containing Opn4 mRNA | Percentage of Opn4 mRNA-positive RGCs containing tracer |
|---|---|---|---|---|---|
| SCN contra | |||||
| 2346 | 5 | 56 | 6.9 | 77.8 | 67.5 |
| 2348 | 5 | 48 | 5.9 | 77.4 | 69.6 |
| 2350 | 5 | 56 | 6.9 | 54.9 | 80.0 |
| 2353 | 5 | 80 | 9.9 | 62.5 | 77.7 |
| Mean | 7.4 ± 0.86 | 68.1 ± 5.7 | 73.7 ± 3.0 | ||
| SCN ipsi | |||||
| 2346 | 5 | 46 | 5.7 | 73.0 | 58.2 |
| 2348 | 5 | 61 | 7.5 | 76.3 | 58.7 |
| 2350 | 5 | 75 | 9.3 | 55.6 | 87.2 |
| 2353 | 5 | 83 | 10.3 | 83.0 | 76.9 |
| Mean | 8.2 ± 1.0 | 72.0 ± 5.8 | 70.2 ± 7.1 | ||
| vSPZ contra | |||||
| 2548 | 96 | 27 | 0.18 | 79.4 | 2.8 |
| 2550 | 96 | 8 | 0.05 | 57.1 | 0.82 |
| 2567 | 114 | 29 | 0.17 | 85.3 | 1.5 |
| Mean | 0.14 ± 0.04 | 73.9 ± 8.6 | 1.68 ± 0.57 | ||
| VLPO contra | |||||
| 2444 | 151 | 14 | 0.06 | 56.0 | 0.61 |
| 2445 | 176 | 24 | 0.09 | 77.4 | 0.67 |
| 2485 | 161 | 11 | 0.05 | 57.9 | 0.53 |
| Mean | 0.07 ± 0.01 | 63.8 ± 6.8 | 0.60 ± 0.04 | ||
| PTA contra | |||||
| 2436 | 5 | 51 | 6.6 | 9.0 | 70.8 |
| 2437 | 5 | 68 | 8.8 | 8.3 | 73.9 |
| 2438 | 5 | 23 | 3.0 | 18.7 | 46.0 |
| 2439 | 5 | 48 | 6.2 | 9.0 | 73.8 |
| Mean | 6.2 ± 1.2 | 11.3 ± 2.5 | 66.1 ± 6.8 | ||
| LGN ipsi | |||||
| 2454 | 5 | 19 | 2.5 | 35.2 | 17.8 |
| 2455 | 5 | 11 | 1.4 | 19.6 | 16.4 |
| 2457 | 5 | 18 | 2.3 | 34.0 | 18.8 |
| Mean | 2.1 ± 0.33 | 29.6 ± 5.0 | 17.6 ± 0.68 |
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Data for the eye contralateral to the injection site are shown for SCN-, vSPZ-, VLPO-, and PTA-injected animals, and results for the ipsilateral eye are shown for SCN- and LGN (IGL)-injected animals (see Results). The column labeled double-labeled RGCs indicates the raw total of cells that contain both retrograde tracer and melanopsin mRNA, but cell counts in the column labeled double-labeled RGCs per section have been corrected using the Abercrombie method (tissue thickness was 20 μm for SCN-injected animals and 25 μm for all other animals). Mean soma diameter of retrogradely labeled RGCs (in μm): SCN, 12.34 ± 0.47; vSPZ, 13.1 ± 0.54; VLPO, 12.7 ± 0.71; PTA, 13.42 ± 0.43; LGN (IGL), 13.31 ± 0.62. The column labeled percentage of tracer-positive RGCs containing Opn4 mRNA indicates the percentage of retrogradely labeled cells that contain melanopsin transcript. Conversely, percentage of Opn4 mRNA-positive RGCs containing tracer indicates the percentage of melanopsin-containing RGCs that are also immunoreactive for the retrograde tracer. The percentage colocalization was determined for each animal, and the mean values for each group of injected animals are reported below each group of injected animals. The SEM is shown for all values.
