Table 2.
Number of identified single-labelled, c-Fos+ neurons, or double-labelled, ChAT+/c-Fos+, Ser+/c-Fos+, TH+/c-Fos+, and GAD+/c-Fos+ neurons in the cholinergic, serotonergic, noradrenergic and adjacent central gray cell groups of the pontomesencephalic tegmentum in PSC, PSD, and PSR groups
| Nucleus | c-Fos+2-a | ChAT+/c-Fos+2-b | ||||||
|---|---|---|---|---|---|---|---|---|
| PSC | PSD | PSR | F | PSC | PSD | PSR | F | |
| LDTg | 61.25 ± 10.08 | 63.25 ± 12.94 | 106.38 ± 14.89*,† | ‡ | 42.13 ± 3.53 | 23.33 ± 5.93* | 53.83 ± 6.57† | ‡ |
| LDTgV | 33.75 ± 5.48 | 53.13 ± 5.23 | 67.75 ± 6.42* | ‡ | 20.88 ± 3.19 | 15.00 ± 1.53 | 16.50 ± 1.80 | |
| PPTgM | 113.25 ± 34.79 | 83.50 ± 7.63 | 96.25 ± 7.64 | 12.75 ± 3.43 | 9.83 ± 0.44 | 20.83 ± 4.34*,† | ‡ | |
| PPTgL | 177.50 ± 57.47 | 153.00 ± 18.06 | 257.50 ± 77.33 | 24.63 ± 4.71 | 27.17 ± 4.92 | 32.00 ± 1.00 | ||
| Sum2-c | 385.63 ± 89.83 | 352.88 ± 22.38 | 527.88 ± 102.62*,† | ‡ | 100.25 ± 12.83 | 75.35 ± 11.32 | 123.17 ± 12.41*,† | ‡ |
| DR | 37.50 ± 3.75 | 64.50 ± 28.12 | 113.63 ± 40.11*,† | ‡ | ||||
| MR | 267.75 ± 109.45 | 213.33 ± 43.91 | 109.33 ± 32.09 | |||||
| Sum2-c | 305.25 ± 109.54 | 224.50 ± 44.37 | 225.00 ± 25.51 | |||||
| LC | 28.24 ± 4.50 | 32.75 ± 3.43 | 15.00 ± 2.38*,† | ‡ | ||||
| SubCA | 63.00 ± 12.99 | 97.50 ± 15.60 | 61.00 ± 13.37 | |||||
| Sum2-c | 91.25 ± 16.67 | 130.25 ± 17.19 | 76.00 ± 14.01† | ‡ | ||||
| rCG | 159.00 ± 43.54 | 142.75 ± 11.81 | 238.00 ± 30.05*,† | ‡ | ||||
| cCG | 124.00 ± 27.58 | 120.25 ± 16.47 | 161.00 ± 19.79 | |||||
| Sum2-c | 283.00 ± 63.02 | 263.00 ± 27.57 | 399.00 ± 48.96*,† | ‡ | ||||
| Ser+/c-Fos+2-b | TH+/c-Fos+2-b | GAD+/c-Fos+2-b | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| PSC | PSD | PSR | F | PSC | PSD | PSR | F | PSC | PSD | PSR | F |
| 29.50 ± 10.77 | 21.00 ± 2.78 | 87.33 ± 13.37*,† | ‡ | ||||||||
| 11.67 ± 2.96 | 18.17 ± 5.29 | 31.00 ± 6.76*,† | ‡ | ||||||||
| 26.17 ± 11.61 | 19.33 ± 2.59 | 44.40 ± 9.68*,† | ‡ | ||||||||
| 29.17 ± 2.32 | 30.00 ± 2.65 | 57.33 ± 21.36*,† | ‡ | ||||||||
| 96.33±17.46 | 88.50 ± 4.36 | 220.07 ± 46.25*,† | ‡ | ||||||||
| 24.87 ± 3.60 | 29.17 ± 5.96 | 12.83 ± 0.88*,† | ‡ | 10.50 ± 2.75 | 7.67 ± 2.60 | 29.17 ± 4.34*,† | ‡ | ||||
| 33.75 ± 7.62 | 39.75 ± 11.25 | 14.17 ± 6.93*,† | ‡ | 44.67 ± 14.17 | 38.00 ± 10.61 | 104.83 ± 35.55*,† | ‡ | ||||
| 58.50 ± 11.26 | 67.17 ± 8.59 | 27.00 ± 7.29*,† | ‡ | 53.50 ± 12.86 | 45.67 ± 8.82 | 134.00 ± 31.27*,† | ‡ | ||||
| 9.00 ± 0.74 | 12.13 ± 2.59* | 7.67 ± 0.60† | ‡ | ||||||||
| 1.38 ± 0.47 | 2.50 ± 0.71 | 0.50 ± 0.50† | ‡ | 31.33 ± 5.21 | 28.00 ± 4.00 | 51.00 ± 10.82*,† | ‡ | ||||
| 10.37 ± 0.47 | 14.63 ± 2.95* | 8.00 ± 0.29† | ‡ | ||||||||
| 42.67 ± 11.67 | 24.50 ± 6.81 | 122.50 ± 12.66*,† | ‡ | ||||||||
| 47.50 ± 22.53 | 43.83 ± 8.25 | 74.00 ± 9.47*,† | ‡ | ||||||||
| 90.17 ± 32.76 | 68.33 ± 11.11 | 196.50 ± 13.44*,† | ‡ | ||||||||
Cell counts were taken from five adjacent series of sections cut at 25 μm and collected at 200 μm intervals.
c-Fos+ cell numbers correspond to the mean ± SEM of the total number of single-labelled c-Fos+ cells counted unilaterally per nucleus in two sections (at ∼A 0.5 and ∼A 0.1) for all rostral structures or in one section at (∼P 0.3) for LC, SubCA, and cCG. Sum refers to the mean ± SEM of the total unilateral cell count for nuclei of the cholinergic, serotonergic, noradrenergic, or central gray cell groups.
Double-labelled cell numbers correspond to the mean ± SEM of the total number of cells counted bilaterally and averaged for the two sides in sections extending through the cholinergic, serotonergic, noradrenergic, or neighboring central gray cell groups. Double-labelled cholinergic and serotonergic cells were counted through the major extent of their rostrocaudal distribution, which varies according to the specific nucleus (Ford et al., 1995). Double-labelled noradrenergic cells were counted through the rostral to middle levels of their distribution. Double-labelled GABAergic cells were counted on adjacent sections at corresponding levels in each cell group and nucleus (except the LC) and also in rostral and caudal central gray areas neighboring the DR and LC, respectively. Accordingly, counts were performed in the following number of sections for each nucleus. ChAT+/c-Fos+ cells or GAD+/c-Fos+ cells were counted: in four sections for the LDTg (between ∼A 0.7 and ∼A 0.1) and the PPTgM (between ∼A 0.9 and ∼A 0.3) and in five sections for LDTgV (between ∼A 0.7 and ∼P 0.1) and PPTgL (between ∼A 1.1 and ∼A 0.3). Ser+/c-Fos+ or GAD+/c-Fos+ cells were counted in five sections for the DR and MR (between ∼A 0.7 and ∼P 0.1). TH+/c-Fos+ cells were counted in three sections for the LC (between ∼P 0.1 and ∼P 0.5), and TH+/c-Fos+ or GAD+/c-Fos+ cells in two sections for the SubCA (at ∼P 0.1 and ∼P 0.3). GAD+/c-Fos+ cells were also counted in five sections for the rCG (between ∼A 0.9 and ∼A 0.1) and in three sections for the cCG (between ∼P 0.1 and ∼P 0.5). Sum refers to the mean ± SEM of the total (average) unilateral cell count for corresponding nuclei of the cholinergic, serotonergic, noradrenergic, or central gray cell groups.
Statistical tests of the variation in cell numbers for the sum of neurons in cholinergic, serotonergic, noradrenergic, or central gray cell groups were performed using one-way ANCOVAs with condition as grouping factor and nucleus, section, and animal as covariates.
For Sum, a significant main effect for condition is indicated by
p ≤ 0.05 in the column under F. With a significant main effect for sum, individual ANCOVAs were performed for each nucleus with condition as grouping factor and section and animal as covariates. For individual nuclei, a significant main effect for condition is indicated by
p ≤ 0.05 in the column under F. For both sum and individual nuclei with significant main effects of condition, post hoc tests between individual conditions were performed using Fisher’s pairwise comparisons. A significant difference with respect to PSC is indicated by * p ≤ 0.05 and with respect to PSD by
p ≤ 0.05.