Skip to main content
. 2020 May 28;23(6):101203. doi: 10.1016/j.isci.2020.101203

Figure 1.

Figure 1

Conditional KO of PTPσ Impairs Excitatory Synapse Development and Transmission in Cultured Hippocampal Neurons

(A) PTPσ cKO in cultured hippocampal neurons specifically reduces excitatory synapse density. Double immunofluorescence analysis of MAP2 (blue) and the indicated synaptic markers (red) in mature cultured neurons (DIV14) derived from PTPσf/f mice infected with lentiviruses expressing ΔCre or Cre at DIV3. Synaptic markers assayed included surface GluA1 (sGluA1), total GluA1, Shank, and VGLUT1 as excitatory synaptic markers, and surface GABAAγ2 (sGAγ2), total GABAAγ2, Gephyrin (Gphn), and GAD67 as inhibitory synaptic markers. Scale bar: 10 μm.

(B and C) Quantification of images in (A), measuring the density (B) and area (C) of the indicated synaptic marker puncta. Data are means ± SEMs (n denotes number of analyzed neurons; ΔCre/PTPσ cKO/sGluA1, n = 16; Cre/PTPσ cKO/sGluA1, n = 17; ΔCre/PTPσ cKO/GluA1, n = 16; Cre/PTPσ cKO/GluA1, n = 15; ΔCre/PTPσ cKO/Shank, n = 16; Cre/PTPσ cKO/Shank, n = 16; ΔCre/PTPσ cKO/VGLUT1, n = 15; Cre/PTPσ cKO/VGLUT1, n = 16; ΔCre/PTPσ cKO/sGABAARγ2, n = 15; Cre/PTPσ cKO/sGABAARγ2, n = 15; ΔCre/PTPσ cKO/GABAARγ2, n = 16; Cre/PTPσ cKO/GABAARγ2, n = 16; ΔCre/PTPσ cKO/Gephyrin, n = 16; Cre/PTPσ cKO/Gephyrin, n = 15; ΔCre/PTPσ cKO/GAD67, n = 16; and Cre/PTPσ cKO/GAD67, n = 16. Mann-Whitney U test; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001).

(D and E) Representative mEPSC traces (D) and quantification of frequencies, amplitudes, and kinetics (E) of mEPSCs recorded from hippocampal cultured neurons derived from PTPσf/f mice infected with lentiviruses expressing inactive (ΔCre) or active (Cre) Cre recombinase. Data are means ± SEMs (n denotes number of analyzed neurons; ΔCre, 20 and Cre, 15; unpaired t test; ∗p < 0.05).

(F and G) Representative mIPSC traces (F) and quantification of frequencies, amplitudes, and kinetics (G) of mIPSCs recorded from hippocampal cultured neurons derived from PTPσf/f mice infected with lentiviruses expressing ΔCre or Cre. Data are means ± SEMs (n denotes number of analyzed neurons; ΔCre, 18 and Cre, 17; unpaired t test)