Quantification of neurite outgrowth induced by PACAP38 in variant PC12 cell lines
| PC12 cell line | Neurite length (× cell diameter) | % Neurite-bearing cells (mean ± standard error) | No. of fields evaluated | No. of cells/field |
|---|---|---|---|---|
| Wild-type | 2–3 | 80.7 ± 7.51-a | 12 | 600 |
| nnr5 | 1–2 | 52.8 ± 8.91-a | 9 | 160 |
| 6.24 | 2–4 | 85.1 ± 8.11-a | 12 | 220 |
| SrcDN2 | 1–2 | 64.6 ± 7.31-a | 9 | 90 |
| RasDN6 | 1–2 | 41.0 ± 8.61-a | 15 | 140 |
| M-M17-26 | 1–2 | 54.7 ± 10.61-a | 12 | 170 |
The following PC12 cell lines were seeded at low density (1000–5000 cells/dish) on collagen/poly-l-lysine-coated tissue culture dishes: wild-type, nnr5 (NGF-nonresponsive), 6.24 (p140trkoverexpressing), SrcDN2 (induced dominant negative Src), RasDN6 (induced dominant negative Ras), and M-M17-26 (constitutive dominant negative Ras). Cultures were incubated 48 hr in the absence or presence of PACAP38, examined by light microscopy, and photographed. Neurite length, measured directly from photographs, is expressed as a function of cell diameter. In untreated, control cultures for each cell line, <5% of the cells exhibit neuritic processes with an overall length less than one cell diameter.
↵1-a Statistically significant (p< 0.001) compared with neurite outgrowth in untreated control cultures by analysis of variance.