Table 1.
Effect of Ror1 and Ror2 suppression by RNAi on neurite elongation in hippocampal neurons
| Days in culture | Treatment siRNA duplex) | Dose (nM) | Primary minor process length (μm) | % Branched primary minor processes | Primary axon length (μm) | Number of branches/axon |
|---|---|---|---|---|---|---|
| 1 | None | – | 37±1 | 45±3 | – | – |
| 1 | Non-specific siRNA | 100 | 37±2 | 38±3 | – | – |
| 1 | siRNA Ror1 SCR | 100 | 35±0 | 47±2 | – | – |
| 1 | siRNA Ror1 MUT | 100 | 37±1 | 47±4 | – | – |
| 1 | siRNA Ror2 SCR | 100 | 35±0 | 44±3 | – | – |
| 1 | siRNA Ror2 MUT | 100 | 34±1 | 43±4 | – | – |
| 1 | siRNA Ror1 | 100 | 28±1* | 26±2* | – | – |
| 1 | siRNA Ror2 | 100 | 30±1* | 32±2* | – | – |
| 2 | None | – | 46±2 | 44±3 | 190±6 | 2.2±0.1 |
| 2 | Non-specific siRNA | 100 | 43±2 | 37±3 | 181±7 | 1.7±0.1 |
| 2 | siRNA Ror1 SCR | 100 | 41±2 | 51±4 | 164±5 | 2.7±0.2 |
| 2 | siRNA Ror1 MUT | 100 | 40±2 | 44±3 | 192±8 | 2.2±0.1 |
| 2 | siRNA Ror2 SCR | 100 | 41±2 | 58±5 | 165±5 | 2.3±0.2 |
| 2 | siRNA Ror2 MUT | 100 | 39±2 | 44±3 | 186±6 | 1.8±0.1 |
| 2 | siRNA Ror1 | 100 | 35±2* | 25±2* | 241±8* | 1.4±0.1* |
| 2 | siRNA Ror1 | 50 | 33±2* | 34±4 | 222±10* | 1.4±0.1* |
| 2 | siRNA Ror1 | 10 | 33±3* | 35±4 | 226±10* | 1.4±0.1* |
| 2 | siRNA Ror1 | 1 | 38±2 | 41±4 | 192±9 | 2.1±0.2 |
| 2 | siRNA Ror2 | 100 | 33±2* | 28±3* | 210±6* | 1.5±0.1* |
| 2 | siRNA Ror2 | 50 | 32±2* | 34±3 | 207±8 | 1.4±0.1* |
| 2 | siRNA Ror2 | 10 | 34±2* | 36±4 | 209±10 | 1.3±0.1* |
| 2 | siRNA Ror2 | 1 | 34±2* | 35±4 | 201±8 | 1.6±0.1 |
E16 hippocampal cultures were plated at 250,000 cells/dish and allowed to develop for 1 (stage II) or 2 (stage III) days. Cells were fixed and stained with a tubulin antibody (clone DMIA). Neuritic processes from randomly selected cells were viewed by phase microscopy using a digital camera and traced on the screen. Neuritic length was determined using Metamorph Image Analysis Software. At least 90 cells from three independent experiments were analyzed for each condition. Results are expressed as mean±s.e.m.
P<0.01 compared to measurements in all controls.
Non-specific siRNA, siRNA targeted against a non-mammalian gene; siRNA SCR, siRNA containing a scrambled sequence; siRNA MUT, siRNA containing a 2 base pair change.