Fig. 2: MPS II neurons exhibited anomalies in neurite morphology, NEFL gene expression, and action potential. | Cell Death & Disease

Fig. 2: MPS II neurons exhibited anomalies in neurite morphology, NEFL gene expression, and action potential.

From: Mature neurons from iPSCs unveil neurodegeneration-related pathways in mucopolysaccharidosis type II: GSK-3β inhibition for therapeutic potential

Fig. 2: MPS II neurons exhibited anomalies in neurite morphology, NEFL gene expression, and action potential.

A Neurite morphometrics of mature cortical neurons. The 15-week MPS II, HC, and ISO neurons were infected with AAV8-hSyn-EGFP. Neuron morphology was reconstructed using Neurolucida. The analyzed neuron morphology parameters included soma size, segments, bifurcation nodes, terminal ends, and total branch lengths. N = 3 HC clones, N = 4 ISO clones from 2 patients, and N = 12 MPS II clones from 4 patients. B Flow cytometry analyses demonstrated decreased NEFL, Ankyrin G, and Spectrin protein levels in 15-week MPS II neurons. N = 6 HC clones, N = 6 differentiations from 2 patients’ 4 ISO clones, and N = 6 MPS II clones from 4 patients. C Abnormal action potentials (APs) in mature MPS II neurons compared to HC and ISO neurons. The APs of cortical neurons (differentiated for 18 weeks from control and MPS II iPSCs) in response to increasing current steps (−50 to 150 pA) were measured using current-clamp recordings. A decreased response rate to current injections in MPS II neurons compared to those of HC and ISO neurons (upper left panel). N = 14 cells of 4 differentiations of HC-1, N = 15 cells from 4 differentiations of ISO-4, and N = 24 cells from 3 differentiations of MPS II 4-1. ***p < 0.005, analyzed by chi-square test. Representative traces depicted the changes of AP morphology in MPS II neurons compared with HC and ISO neurons. Scale bars: 10 ms (x-axis) and 10 mV (y-axis). Histograms show the differences of AP parameters between MPS II neurons and HC or ISO neurons, including AP threshold, AP spike height, AP half-width, after-hyperpolarization (AHP), the slope of 10–90% rise, and the slope of 90–10% decay. Each data point represents an individual responsive neuron. N = 12 cells from 4 differentiations of HC-1, N = 13 cells from 4 differentiations of ISO-4, and N = 6 cells from 3 differentiations of MPS II 4-1. Data represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005, analyzed by Mann–Whitney U test. Cell clone details are in Supplementary Table 1.

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