Table 2.
Effect of Ror1 and Ror2 suppression by means of antisense oligonucleotides on neurite elongation in hippocampal neurons
| Days in culture | Treatment (oligonucleotide) | Dose (μM) | Primary minor process length (μm) | % Branched primary minor processes | Primary axon length (μm) | Number of branches/axon |
|---|---|---|---|---|---|---|
| 1 | None | – | 35±3 | 35±4 | – | – |
| 1 | Sense Ror1 (+80+97) | 50 | 33±7 | 34±2 | – | – |
| 1 | Sense Ror2 (+24+41) | 50 | 32±7 | 35±2 | – | – |
| 1 | Antisense Ror1 (+80+97) | 50 | 25±9*,† | 15±3*,† | – | – |
| 1 | Antisense Ror2 (+24+41) | 50 | 26±8*,† | 14±3*,† | – | – |
| 2 | None | – | 41±2 | 60±1 | 100±3 | 3.3±0.3 |
| 2 | Sense Ror1 (+80+97) | 50 | 34±2 | 64±1 | 103±7 | 2.1±0.2 |
| 2 | Sense Ror2 (+24+41) | 50 | 38±2 | 60±1 | 109±7 | 2.3±0.2 |
| 2 | Antisense Ror1 (+80+97) | 50 | 28±2*,† | 30±1*,† | 155±9*,† | 1.7±0.2* |
| 2 | Antisense Ror1 (+80+97) | 25 | 29±2*,† | 30±1*,† | 139±8*,† | 1.8±0.3* |
| 2 | Antisense Ror1 (+80+97) | 12 | 31±2* | 23±1*,† | 130±7*,† | 1.6±0.2* |
| 2 | Antisense Ror2 (+24+41) | 50 | 28±2*,† | 24±1*,† | 153±8*,† | 1.4±0.9* |
| 2 | Antisense Ror2 (+24+41) | 25 | 28±2*,† | 29±1*,† | 130±10* | 1.5±0.2* |
| 2 | Antisense Ror2 (+24+41) | 12 | 32±2*,† | 31±1*,† | 109±7 | 1.6±0.2* |
E16 hippocampal cultures were plated at 75,000 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-contrast microscopy using a digital camera and traced onscreen. Neuritic length was determined using Metamorph Image Analysis Software. Sixty cells from three independent experiments were analyzed for each condition. Results are expressed as mean±s.e.m.
P<0.01compared to measurements in the untreated control.
P<0.01 compared to measurements in the corresponding sense control.