Fig. 5: During early CNS development impaired release of TWEAK from GABABR-deficient OPCs mitigated programmed cell death of interneurons. | Nature Communications

Fig. 5: During early CNS development impaired release of TWEAK from GABABR-deficient OPCs mitigated programmed cell death of interneurons.

From: Impaired bidirectional communication between interneurons and oligodendrocyte precursor cells affects social cognitive behavior

Fig. 5

a Experimental schedule. b Fluorescent micrographs of excitatory (CtBP-interacting protein (CTIP)+, magenta) and inhibitory (parvalbumin (pv)+, magenta) interneurons co-stained with the apoptotic marker cleaved caspase 3 (CC-3, green) in p5 and p7 mPFC. c Quantification of apoptotic neurons (CC-3+) co-expressing the interneuron marker glutamate decarboxylase 67 (GAD67) and PV or CTIP and T-box brain transcription factor 1 (TBR1) as markers for excitatory neurons. (GAD67: p5-ctl = 3 mice, p5-cKO = 4 mice; p7-ctl = 3 mice, p7-cKO = 5 mice) (PV: p5-ctl = 5 mice, p5-cKO = 3 mice; p7-ctl = 3 mice, p7-cKO = 5 mice) (CTIP: p5-ctl = 7 mice, p5-cKO = 4 mice; p7-ctl = 9 mice, p7-cKO = 3 mice) (TBR1: p5-ctl = 4, p5-cKO = 4; p7-ctl = 4, p7-cKO = 4 mice) (two-sided unpaired t-tests). d Relative mRNA level of TNF like weak inducer of apoptosis (TWEAK) in the medial prefrontal cortex (mPFC), magnetic-activated cell sorted (MACs)-OPCs from ctl and cKO mice as well as Oli-neu cells treated with or without 20 µM CGP 55845 (tissue-ctl = 4 mice, tissue-cKO = 6 mice; MACs-ctl = 8 mice, MACs-cKO = 4 mice; Oli-neu: n = 3 independent experiments; two-sided unpaired t-tests). e Double-immunostaining of TWEAK receptor (TWEAKR, magenta) and PV (green) in the control mPFC at p5. Arrows indicate co-expression of TWEAKR and PV. f Experimental design for in vitro studies. g Immunolabeling of TWEAKR (magenta) on 14 days in vitro (DIV) PV+ interneurons (green). h Immunostaining of apoptotic primary PV+ neurons with CC-3 (green) and PV (magenta) after being treated with conditional medium (CM) of Oli-neu cells co-treated with or without TWEAKR antagonist L524-0366 (L524, 20 µM). White arrows indicate PV+ cells co-expressing CC-3. i Percentage of apoptotic PV+ interneurons of all PV+ interneurons in conditioned medium and treated with a competitive TWEAKR inhibitor (n = 3 independent experiments from distinct samples, one-way ANOVA, Turkey’s multiple comparison). Data are shown as mean ± SEM in c, d, and i. Source data are provided as a Source Data file.

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