Figure 2.

Differentiated iNSCs Functionally Integrate into the Existing Neuronal Circuitry

(A) An immunohistological analysis of the iNSC-derived cells that differentiated into glutamatergic cells 6 months after transplantation into the cortex and the hilus.

(B) Immunohistological stainings indicated a differentiation into GABAergic subtypes in both regions. The left image of the upper panel of the cortex transplantation and the dentate gyrus transplantation represents the MIP of a confocal z stack. Dashed lines indicate the region of magnification.

(C) iNSC-derived neurons matured as indicated by the positive immunohistological staining for NEUN, integrated into cortex, and showed a complex morphology similar to the typical neuronal appearance of a pyramidal subtype. The upper panel represents the MIP of a confocal z stack. The lower panel represents a higher magnification of a single plane of the cell body.

(D) Creating a 3D surface of the confocal z stacks of the cell shown in Figure 2C reveals a 3D spatial orientation. Arrows indicate the development of dendritic spines with typical head-neck structures.

(E) The 3D surface of confocal z stacks from the immunohistological staining of the presynaptic marker SYNAPTOPHYSIN indicated the formation of synaptic connections with endogenous cells. Dashed lines indicate the region of magnification. Arrows indicate direct contact between the neurite (green) and multiple presynapses (red) of the surrounding endogenous cells.

Nuclei were counterstained with Hoechst.