Article Figures & Data
Figures
- Figure 1.
Experimental design of experiment 1. T@P and sham-T@P males arrived four weeks before NoT@P and sham-NoT@P males to control for the age of shipping and environment during puberty. Two to 7 d after the NoT@P and sham-NoT@P arrived, males were either GDX or sham-GDX during adulthood (P56; T@P and sham-T@P) or prepubertally (P28; NoT@P and sham-NoT@P). Four weeks later, T@P and NoT@P males received testosterone (T)-filled capsules and sham males received empty (blank) capsules of the same size. Sexual behavior testing began two weeks later.
- Figure 2.
Rate of ectopic mounting is dependent on pubertal testosterone. T@P males had significantly fewer ectopic mounts per minute compared to NoT@P males. Bars represent mean (±SEM); numbers on bars indicate sample size. *Main effect of pubertal testosterone, p ≤ 0.05.
- Figure 3.
The effects of pubertal testosterone and sexual experience on latency to mount, intromit, and ejaculate and number of intromissions to ejaculation. Mount latency: there was a main effect (ME) of sexual experience on mount latency with sexually experienced males having shorter latencies to mount compared to sexually naive males. Intromission latency: There was an ME of sexual experience on intromission latency with sexually experienced males having shorter latencies to intromit compared to sexually naive males. Ejaculation latency: there was a pubertal testosterone × sexual experience interaction on ejaculation latency with sexually naive NoT@P males having a longer latency to ejaculate compared to sexually naive T@P males. This effect was not seen in sexually experienced males. Intromissions to ejaculate: there was an ME of sexual experience for intromissions to ejaculate with sexually experienced males having less intromissions to achieve ejaculation compared to sexually naive males. Bars represent mean (±SEM); numbers on bars indicate sample size. +Interaction between pubertal testosterone and sexual experience, p ≤ 0.05.
- Figure 4.
Number of ΔFosB-ir cells in IL is dependent on pubertal testosterone and sexual experience. A, Brain atlas (Morin and Wood, 2001) representation of a coronal section containing the mPfC. B, Photomicrographs of drawn contours of the mPfC onto immunohistochemically-treated tissue sections at 4× objective. The mPfC included the anterior Cg1, PrL, and IL cortices; scale bar = 250 µm. C, The 2 × 2 panel of photomicrographs below the bar graph are representative images of ΔFosB-ir in the IL for the specified group of males; scale bars = 25 µm. D, In the CgL and PrL, there were no effects or interactions of pubertal testosterone and sexual experience on ΔFosB-ir cells, respectively. In the IL, there was an interaction between pubertal testosterone and sexual experience on ΔFosB-ir cells with sexual experienced T@P males having significantly more ΔFosB-ir cells compared to sexually naive T@P males. There were no significant differences in ΔFosB-ir cells as a function of sexual experience within NoT@P males. Bars represent mean (±SEM); numbers on bars indicate sample size. +Interaction between pubertal testosterone and sexual experience, p ≤ 0.05.
- Figure 5.
Number of ΔFosB-ir cells in the NAc core and shell is dependent on pubertal testosterone and sexual experience. A, Brain atlas (Morin and Wood, 2001) representation of a coronal section containing the NAc. B, Photomicrographs of drawn contours of the NAc onto immunohistochemically-treated tissue sections. The NAc included the shell and core. LV = lateral ventricle; ac = anterior commissure. Scale bar = 250 µm. C, In the core, there was an interaction between pubertal testosterone and sexual experience on ΔFosB-ir cells with sexual experienced T@P males having significantly more ΔFosB-ir cells compared to sexually naive T@P males. There were no significant differences in ΔFosB-ir cells as a function of sexual experience within NoT@P males. In the shell, there was a main effect (ME) of sexual experience on ΔFosB-ir cells with sexually experienced males having more ΔFosB-ir expression compared to sexually naive males. Bars represent mean (±SEM); numbers on bars indicate sample sizes. +Interaction between pubertal testosterone and sexual experience, p ≤ 0.05.
- Figure 6.
Visualization of GFP to verify injection site and extent of infected cells in the IL. A, Boxes of representative injection sites in the IL for NoT@P-ΔFosB males over coronal atlas diagram (Morin and Wood, 2001). B, Photomicrograph of GFP overexpression in a NoT@P-ΔFosB male; scale bar = 250 µm. C, Photomicrograph of GFP overexpression in a NoT@P-ΔFosB male; scale bar = 100 µm.
- Figure 7.
Overexpression of ΔFosB in the IL decreases the rate of ectopic mounting in NoT@P males. NoT@P-ΔFosB males (n = 6) had significantly less ectopic mounts per minute compared to NoT@P-GFP males (n = 9). T@P-GFP males (n = 7) did not differ from either group in rate of ectopic mounting. Bars represent mean (±SEM). *Main effect of experimental group, p ≤ 0.05.
- Figure 8.
The effects of ΔFosB overexpression in the IL and sexual experience on latency to mount, intromit, and ejaculate and number of intromissions to ejaculation. For mount latency, there was a pubertal testosterone × sexual experience interaction with sexually naive NoT@P-ΔFosB males having a longer latency to mount compared to sexually experienced NoT@P-ΔFosB males. This effect of sexual experience was not found in T@P-GFP or NoT@P-GFP males. For intromission latency and ejaculation latency, there was a main effect of sexual experience with sexually experienced males having shorter latencies to mount and intromit compared to sexually naive males. For intromissions to ejaculate, there was a main effect (ME) of sexual experience with sexually experienced males having less intromissions to achieve ejaculation compared to sexually naive males. Bars represent mean (±SEM); numbers on bars indicate sample size. +Interaction between experimental group and sexual experience, p ≤ 0.05.
- Figure 9.
Both sexual experience and ΔFosB increase dendritic spines in vmPFC. A, AAV-GFP was injected into the IL of naive (n = 22) or sexually experienced (n = 21) males, and AAV-GFP-ΔFosB was injected into the IL of naive males (n = 22). Immunofluorescence using a GFP antibody reveals spines of IL pyramidal neurons in all three groups. B, Thin spine density was increased by ΔFosB overexpression. Stubby spine density was not affected by either sexual experience nor ΔFosB overexpression. Mushroom spine density increased with sexual experience and showed a trend to increase by ΔFosB overexpression. Overall, total spine density was increased by ΔFosB overexpression. Bars represent mean (±SEM). *Main effect of experimental group, p < 0.05.
Tables
Plasma testosterone (ng/ml) Group Sexual experience Naive Experienced T@P 2.77 ± 1.01 3.54 ± 1.66 NoT@P 2.77 ± 1.21 3.68 ± 1.30 Sham-T@P 1.71 ± 0.72 2.59 ± 0.78 Sham-NoT@P 1.67 ± 0.88 2.57 ± 0.72 Figure Independent variable(s) Dependent variable Statistics (*significant) NA Sexual experience Latency to mount F(1,24) = 14.65; p = 0.001* Latency to intromit F(1,24) = 14.33; p = 0.001* Latency to ejaculate F(1,24) = 31.27; p = 0.001* Intromissions to ejaculation F(1,17) = 12.24; p = 0.003* ΔFosB in the IL F(1,23) = 4.45; p = 0.046* Figure Independent variable(s) Dependent variable Statistics (*significant) Post hoc comparison (if appropriate) Post hoc statistics (*significant) 2 Pubertal testosterone Ectopic mounting F(1,27) = 6.75; p = 0.015* Sexual experience F(1,27) = 0.69; p > 0.05 Pubertal testosterone × sexual experience F(1,27) = 0.54; p > 0.05 3 Pubertal testosterone Latency to mount F(1,27) = 0.272; p > 0.05 Sexual experience F(1,27) = 17.43; p < 0.001* Pubertal testosterone × sexual experience F(1,27) = 0.309; p > 0.05 Pubertal testosterone Latency to intromit F(1,27) = 1.01; p > 0.05 Sexual experience F(1,27) = 15.84; p < 0.001* Pubertal testosterone × sexual experience F(1,27) = 2.50; p > 0.05 Pubertal testosterone × sexual experience Latency to ejaculate F(1,27) = 12.70; p = 0.001* Pubertal testosterone in naive males F(1,24) = 10.33; p = 0.004* Pubertal testosterone in experienced males F(1,27) = 0.641; p > 0.05 Pubertal testosterone Intromissions to ejaculation F(1,26) = 0.465; p > 0.05 Sexual experience F(1,26) = 25.00; p < 0.001* Pubertal testosterone × sexual experience F(1,26) = 2.73; p > 0.05 4D Pubertal testosterone × sexual experience ΔFosB in the IL F(1,23) = 10.86; p = 0.003* sexual experience in T@P males F(1,12) = 14.06; p = 0.003* sexual experience in NoT@P males F(1,11) = 0.721; p > 0.05 Pubertal testosterone ΔFosB in the Cg1 F(1,23) = 0.046; p > 0.05 Sexual experience F(1,23) = 0.082; p > 0.05 Pubertal testosterone × sexual experience F(1,23) = 0.66; p > 0.05 Pubertal testosterone ΔFosB in the PrL F(1,23) = 0.133; p > 0.05 Sexual experience F(1,23) = 2.04; p > 0.05 Pubertal testosterone × sexual experience F(1,23) = 2.57; p > 0.05 5C Pubertal testosterone × sexual experience ΔFosB in the NAc core F(1,23) = 5.42; p = 0.029* Sexual experience in T@P males F(1,12) = 9.66; p = 0.009* Sexual experience in NoT@P males F(1,11) = 0.042; p > 0.05 Pubertal testosterone ΔFosB in the NAc shell F(1,23) = 1.60; p > 0.05 Sexual experience F(1,23) = 7.041; p = 0.014* Pubertal testosterone × sexual experience F(1,23) = 2.68; p > 0.05 Figure Independent variable(s) Dependent variable Statistics (*significant) Post hoc comparison (if appropriate) Post hoc statistics (*significant) 7 ΔFosB overexpression Ectopic mounting F(2,22) = 3.94; p = 0.035* NoT@P-ΔFosB vs NoT@P-GFP males p = 0.043* Sexual experience F(1,21) = 1.25; p > 0.05 ΔFosB overexpression × sexual experience F(1,21) = 1.20; p > 0.05 8 ΔFosB overexpression × sexual experience Latency to mount F(2,19) = 3.61; p = 0.047* Sexual experience in NoT@P-ΔFosB males F(1,6) = 11.00; p = 0.015* Sexual experience in T@P-GFP males F(1,6) = 1.61; p > 0.05 Sexual experience in NoT@P-GFP males F(1,7) = 0.61; p > 0.05 ΔFosB overexpression Latency to intromit F(2,20) = 1.12; p > 0.05 Sexual experience F(1,20) = 14.09; p = 0.001* ΔFosB overexpression × sexual experience F(2,20) = 3.06; p > 0.05 ΔFosB overexpression Latency to ejaculate F(2,20) = 2.23; p > 0.05 Sexual experience F(1,21) = 15.20; p < 0.001* ΔFosB overexpression × sexual experience F(2,20) = 1.64; p > 0.05 ΔFosB overexpression Intromissions to ejaculation F(2,21) = 0.87; p > 0.05 Sexual experience F(1,20) = 11.70; p = 0.003* ΔFosB overexpression × sexual experience F(2,20) = 0.06; p > 0.05 Figure Independent variable(s) Dependent variable Statistics (*significant) Post hoc comparison (if appropriate) Post hoc statistics (*significant) 9B Experimental group (naive-GFP, experienced-GFP, naive-ΔFosB ) Thin spines F(2,62) = 1.717; p < 0.001* Naive-ΔFosB vs naive-GFP males p < 0.001* Naive-ΔFosB vs experienced-GFP males p = 0.016* Experimental group (naive-GFP, experienced-GFP, naive-ΔFosB ) Stubby spines F(2,62) = 0.723; p > 0.05 Experimental group (naive-GFP, experienced-GFP, naive-ΔFosB ) Mushroom spines F(2,61) = 4.080; p = 0.022* Naive-GFP vs experienced-GFP males p = 0.029* Experimental group (naive-GFP, experienced-GFP, naive-ΔFosB ) Total spines F(2,62) = 9.359; p < 0.001* Naive-ΔFosB vs naive-GFP males p < 0.001*
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