Fig. 2: Expression of glutamatergic and GABAergic receptor and synaptic plasticity-related genes in neural organoids over course of differentiation. | Communications Biology

Fig. 2: Expression of glutamatergic and GABAergic receptor and synaptic plasticity-related genes in neural organoids over course of differentiation.

From: Human neural organoid microphysiological systems show the building blocks necessary for basic learning and memory

Fig. 2

a Representative immunocytochemistry images of organoids showing postsynaptic marker (HOMER1) and presynaptic marker (SYP) in 8- and 12-week cultures. In composite images, HOMER1 is shown in blue, and SYP is shown in yellow. Scale bars are 100 µm, 50 µm, and 10 µm, respectively. b Presence of inhibitory post-synaptic marker (Gephyrin), pre-synaptic marker (SYN1) and dendrites (MAP2) in 8- and 12-week organoids. In composite images, Gephyrin is shown in blue, SYN1 in yellow, and MAP2 in grey. Scale bars are 100 µm and 50 µm, respectively. For a, and b, all images were taken at 20x, 100x, and 100x + 4x zoom and processed with ImageJ for visualization. c Gene expression of Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha1 (GABRA1), Glutamate Ionotropic Receptor NMDA Type Subunit 1 (GRIN1), Glutamate [NMDA] Receptor Subunit Epsilon-1 (GRIN2A), and Glutamate [NMDA] Receptor Subunit Epsilon-2 (GRIN2B), Glutamate Ionotropic Receptor AMPA Type Subunit 1 (GRIA1), homer scaffold protein 1 (HOMER1) in organoids over the course of differentiation. d Representative immunocytochemistry images of weeks 8 and 12 organoids stained for Neuronal Pentraxin 2 (NPTX2), Activity-Regulated Cytoskeleton-associated protein (ARC), cAMP response element-binding protein (CREB), and Brain-Derived Neurotrophic Factor (BDNF). Scale bar is 100 µm. e Gene expression over the course of differentiation of immediate early genes (IEGs) ARC, BDNF, Neuronal PAS Domain Protein 4 (NPAS4), NPTX2, Fos proto-oncogene AP-1 transcription factor subunit (FOS), and Early Growth Response 1 (EGR1). f Gene expression of synaptic slasticity- related genes: CREB, calcium/calmodulin-dependent protein kinase II A (CAMK2A), Synaptic Ras GTPase-activating protein 1 (SYNGAP1). g Gene expression of synaptic plasticity -related miRNAs. For all gene expression plots, data is shown as a box and whisker plot (with the box extending from the 25th to 75th percentiles) and represented as log2(Fold Change) normalized to NPCs from 2-3 independent experiments with 3 technical replicates each. In all qPCR experiments, ACTB was used as a reference gene.

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