Skip to main content
. 2016 May 2;5:e15106. doi: 10.7554/eLife.15106

Figure 2. NMDA receptors maintain spines during puberty.

Figure 2.

(a) Representative EPSCs (black) and NMDA EPSCs (red) recorded during puberty in WT or α4 KO hippocampus, in some cases during α5 (50 nM L655) or total (120 μM SR95531) GABAR blockade. In all other cases, 200 nM SR95531 was bath applied block synaptic GABARs (Stell and Mody, 2002). Scale, 150 pA, 15 ms. (b) Averaged NMDA/AMPA ratios; ANOVA, F(3,31)=20.21, p=0.0001*, power=1; n=8–10 cells (mice)/group. (Figure 2—source data 1) *p<0.05 vs. WT. (c) Inset, Drug treatment during puberty (PND 35–44) was tested for its effect on post-pubertal spine density (PND 56). Z-stack images, pub and post-pub hippocampus, showing the effects of pubertal vehicle or MK-801 treatment, at a dose shown to increase NMDAR expression (Gao and Tamminga, 1995). Scale, 6 μm. (d) Averaged spine density. Proximal (left): ANOVA, F(2,32)=54.16, p<0.0001*, power=1, n= 11–12 neurons (5 mice)/group; Distal (right)l: ANOVA, F(2,32)=460.1, p<0.0001*, power=1; n=11–12 neurons (5 mice)/group. (Figure 2—source data 2) *p<0.05 vs. other groups. (e) Quantification of spine types. Mushroom, ANOVA, F(2,33)=24.7, p<0.0001*; Stubby, ANOVA, F(2,33)=25.4, p<0.0001*; Thin, ANOVA, F(2,33)=7.66, P=0.002*; power=0.9–1; n=12 neurons (6 mice) /group. *p<0.05 vs. other groups. (Figure 2—source data 3) (f) Z-stack images, pub and post-pub hippocampus, showing the effects of pubertal vehicle or memantine (MEM) treatment, a NMDAR blocker which does not alter NMDAR expression (Cole et al., 2013). Scale, 6 μm. (g) Averaged spine density. *Proximal: ANOVA, F(2,54)=64.12, p<0.0001*, power=1, n=17–20 neurons (4–5 mice) /group; Distal: ANOVA, F(2,56)=33.2, p<0.0001*, power=1, n=19–20 neurons (4–5 mice) /group. (Figure 2—source data 4) *p<0.05 vs. other groups. (h) Quantification of spine types. Mushroom, ANOVA, F(2,45)=89.9, p<0.0001*; Stubby, ANOVA, F(2,45)=9.4, P=0.0004*; Thin, ANOVA, F(2,45)=13.7, P=0.0001*; Bifurcated, ANOVA, F(2,45)=17.7, p<0.0001*; power=1, n=16 neurons (4–5 mice)/group. (Figure 2—source data 5) *p<0.05 vs. other groups.

DOI: http://dx.doi.org/10.7554/eLife.15106.012

Figure 2— source data 1. Figure 2b: NMDA EPSC/ AMPA EPSC ratios recorded from CA1 hippocampal pyramidal cells using whole cell patch clamp techniques for post-pubertal WT (a), α4 KO mice (b), WT hippocampus with SR95531 (c) and WT hippocampus with L-655,708 (L655) (d).
DOI: http://dx.doi.org/10.7554/eLife.15106.013
elife-15106-fig2-data1.xlsx (11.2KB, xlsx)
DOI: 10.7554/eLife.15106.013
Figure 2—source data 2. Figure 2d: Spine counts/20 μm on dendrites of CA1 hippocampal pyramidal cells – proximal (left) and distal (right) for pubertal (Pub), Post-pubertal (Post-pub) – vehicle (VEH), and Post-pub MK-801 (treated with MK-801 during the pubertal period).
DOI: http://dx.doi.org/10.7554/eLife.15106.014
elife-15106-fig2-data2.xlsx (11.2KB, xlsx)
DOI: 10.7554/eLife.15106.014
Figure 2—source data 3. Figure 2e: Spine counts/10 μm for different spine-types on dendrites of CA1 hippocampal pyramidal cells for Figure 1c for Pub, Post-pub vehicle (VEH) and Post-pub MK-801 (treated with MK-801during the pubertal period). .
Spines were identified as: mushroom, stubby, or thin.
DOI: http://dx.doi.org/10.7554/eLife.15106.015
elife-15106-fig2-data3.xlsx (11.4KB, xlsx)
DOI: 10.7554/eLife.15106.015
Figure 2—source data 4. Figure 2g: Spine counts/20 μm on dendrites of CA1 hippocampal pyramidal cells – proximal (left) and distal (right) for α4 KO: pubertal (Pub), Post-pubertal (Post-pub) – vehicle (VEH), and Post-pub memantine (treated with memantine during the pubertal period).
DOI: http://dx.doi.org/10.7554/eLife.15106.016
elife-15106-fig2-data4.xlsx (11.7KB, xlsx)
DOI: 10.7554/eLife.15106.016
Figure 2—source data 5. Figure 2h: Spine counts/10 μm for different spine-types on dendrites of CA1 hippocampal pyramidal cells for Figure 1c for α4 KO: Pub, Post-pub vehicle (VEH) and Post-pub memantine (treated with memantine during the pubertal period). .
Spines were identified as: mushroom, stubby, thin or bifurcated (Bif).
DOI: http://dx.doi.org/10.7554/eLife.15106.017
elife-15106-fig2-data5.xlsx (11.8KB, xlsx)
DOI: 10.7554/eLife.15106.017