Figure 5
Immunocharacterization and electrophysiological properties of GABAergic neuron-like cells (iGNs) induced from hADSCs. (a) Neurofilament-M (NF-M)-immunoreactive neurites radiating from the neurosphere-like cell cluster. Scale bar: 100 μm. (b) Immunocytochemistry analysis of mature neuron (MAP2) and GABAergic neuron (GABA, GAD) expressions. Scale bar: 20 μm. (c) Quantitative analysis of GABAergic neuron and/or mature neuron marker protein expression in at least three different randomly chosen fields. The percentage represents each marker-positive cells over DAPI-positive cells, the total cell number (mean ± SEM). (d) Immunocytochemistry analysis of MAP2 and pre-synaptic marker of GABAergic neuron (vGAT) expressions. Scale bar: 20 μm. (e) Immunocytochemistry analysis of MAP2 and pre-/post-synaptic marker (PSD95/SYP) expressions. Scale bar: 20 μm. (f) Representative trace of AP firing from iGNs recorded in current clamp mode (upper left panel). The protocol for current injection is below the AP trace. The upper right panel shows that applying ramp protocol in iGNs elicited fast inward current followed by depolarization of the holding voltage. Voltage was ramped from −100 to 0 mV gradually for 1 sec. The dotted box indicates magnification of ramp current marked with a star (*). A schematic diagram of the experimental design is shown in the bottom left panel. Spontaneous inhibitory postsynaptic current (IPSC) in the company of glutamatergic receptor blockers (50 M APV and 20 M CNQX) is shown in the bottom right and was not observed in the presence of 10 M bicuculine, a GABAA receptor blocker.
