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. Author manuscript; available in PMC: 2019 Oct 26.
Published in final edited form as: Mol Psychiatry. 2019 Apr 9;24(11):1627–1640. doi: 10.1038/s41380-019-0412-6

Figure 3. Pten knockout neurons have more presynaptic partners.

Figure 3.

(A) Starter retrovirus (RV-GFP/Cre-TVA-Rgp) was injected into the dentate gyrus of 4-week-old Ptenflx/flx or control (WT) mice. Seven weeks later, rabies virus (RbV-EnvA-mCherry) was injected at the same coordinate. Connectivity was analyzed one week after rabies virus injection and eight weeks after birth of starter neurons. (B) Quantification of starter cells positive for both retrovirus and rabies virus (left; WT: 185.3±6.524, Pten KO: 53.33 ± 10.30, t(8)=9.51, p=0.000), and total input cells (rabies virus only; middle; WT: 2389±371.2, Pten KO: 2200±399.9, t(8)=0.326, p=0.753). The ratio of input cells to starter cells (right) was higher in Ptenflx/flx mice than in controls (WT: 12.75±1.581, Pten KO: 42.83±4.828, t(8)=4.88, p=0.001). (C; top) Region-specific connectivity ratios. Pten knockout granule neurons had higher connectivity with the hippocampus (HPC; WT: 8.005±0.8224, Pten KO: 17.33±0.9861, t(8)=6.69, p=0.000), Entorhinal cortex (EC; CEC = caudal EC: WT: 2.675±0.5888, Pten KO: 15.89±2.937, t(8)=3.57, p=0.0073; MEC = medial EC: WT: 0.2425±0.06537, Pten KO: 2.573±0.9835, t(8)=1.89, p=0.095; LEC = lateral EC: WT: 0.3525±0.06981, Pten KO: 2.970±0.5921, t(8)=3.53, p=0.008), and basal forebrain (fb; WT: 0.9975±0.1145, Pten KO: 2.748±0.2484, t(8)=5.41, p=0.001). There was no change in connectivity ratio with the midbrain (Mb; WT: 0.0175±0.004787, Pten KO: 0.1950±0.09095, t(8)=1.56, p=0.157). (C; bottom) Example images showing input neurons (mCherry) in the hippocampus, entorhinal cortex, forebrain, and midbrain. White arrowheads denote starter cells expressing both mCherry and GFP from the starter retrovirus. (D) Inputs from the contralateral hilus. Pten knockout neurons had a greater connectivity ratio with the contralateral hilus (WT: 0.0106±0.004475, Pten KO: 0.2084±0.05082, t(8)=3.11, p=0.015). The majority of inputs onto both WT and Pten knockout neurons from the contralateral hilus were positive for calretinin (CR+; WT: n=3 animals, 83.33±16.67%, Pten KO: n=3 animals, 85.00±7.638%, t(4)=0.091, p=0.932). Data are presented as mean±SEM. Except where otherwise stated, n=4 WT animals and 6 Ptenflx/flx animals. All tests are two-tailed unpaired t-tests. See also Figures S3 and S4.