Figure 5.

Presynaptic input to S1 cortex in Pten^T366A/T366A mice. (A) Schematic showing injection procedure of three viruses (also see the ‘Materials and methods' section). AAV1-Cre was injected (injection 1) into POm and VPm thalamus. Two weeks later, the trans-synaptically labelled S1 cortical neurons expressing Cre, i.e. receiving thalamic input, were targeted with a flex AAV to express the TVA receptor (injection 2). Two weeks after injection 2, the TVA-expressing neurons (GFP+ S1 neurons) were targeted with rabies virus (injection 3). The Cre+, TVA-expressing (green) and mCherry+ expressing (red) neurons were the starter neurons (yellow). One week after injection of rabies, brains were harvested, and local and long-range retrograde-labelled neurons could be observed (red presynaptic neurons). (B) Left graph showing number of (Cre+)/GFP+/mCherrry+ (starter) neurons, right graph showing all presynaptic mCherry+ labelled neurons. (C) Examples with enlarged images showing expression of GPF+ neurons, GFP+/mCherry+ (starter) neurons and presynaptic mCherry+ neurons in wild-type and Pten^T366A/T366A S1 cortices. (D) Graph showing the percentage of presynaptic neurons local in S1 cortex and long-range neurons from other cortical and thalamic areas in Pten^T366A/T366A and wild-type brains. (E) Pie charts showing percentages of dissected presynaptic inputs from long-range areas to S1 in Pten^T366A/T366A and wild-type brains. Graph showing input to motor cortices and thalamus in Pten^T366A/T366A and wild-type brains. (F) Example images of presynaptic neurons in motor cortices, thalamus, S1 cortex contralateral and visual cortices in Pten^T366A/T366A and wild-type brains. Higher magnification of single neurons. Statistical analysis with unpaired t-test, *P < 0.05. Analysis details are provided in Supplementary Table 8. Scale bars = 100 µm, 50 µm in enlarged images.