Fig. 4: PTHS neurons exhibit abnormal electrophysiological properties and gene-expression program. | Nature Communications

Fig. 4: PTHS neurons exhibit abnormal electrophysiological properties and gene-expression program.

From: Transcription Factor 4 loss-of-function is associated with deficits in progenitor proliferation and cortical neuron content

Fig. 4

a Neurons in 2D culture (3 months in neuronal medium) after MAP2 immunostaining (white). b Neurite length and soma area quantification in parent and PTHS neurons in 2D culture. N = 4 patients and respective controls (#1 to #4 pairs indicated below the graphs); 37-94 neurons per group (dots). Means are indicated by the colored lines. c Patch-clamp electrophysiological interrogation of parent and PTHS neurons, showing reduction in spike rate in PTHS neurons after 3 months in neuronal medium (right). Representative traces are shown on the left. N = 10 (parent) or 9 (PTHS) neurons from pair #4. d Sodium (left) and potassium (right) currents in parent (orange) and PTHS (blue) neurons. N = 10 (parent) or 9 (PTHS) neurons. e Relative expression (RT-qPCR) of selected neuronal genes in neurons in 2D culture (2 months in neuronal medium). N = 4 subjects per group (symbols). These genes were selected among the most differentially expressed in N-Glut and N-GABA neurons between parent and PTHS organoids (see Supplementary Data 3 for list of DE genes). f Left: Expression of the same genes shown in (e) in neurons of GbOs, at 8 weeks in vitro. N = 2661 (parent) and 988 (PTHS) cells. Right: For comparison with GbOs, the expression of SLC17A6 (vGLUT2) in CtO neurons is also shown. N = 1401 (parent) and 380 (PTHS). Cells are from pair #4. Symbols in bar graphs indicate parent-patient identities: diamonds, pair #1; squares, pair #2; triangles, pair #3; circles, pair #4. Colors in the figure represent parent (orange) or PTHS (blue) groups. Bar graphs represent mean + SEM. n.s., not statistically significantly different, *p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA with Geisser-Greenhouse correction for repeated measures followed by LSD post-hoc test (b), one-way ANOVA followed by HSD post-hoc test (c and d), two-sample Welch’s t test (e), or Kruskal–Wallis H test (f). Scale bar is 100 μm. See Supplementary Data 1 for sample and effect sizes and exact p-values.

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