Downregulation of an Evolutionary Young miR-1290 in an iPSC-Derived Neural Stem Cell Model of Autism Spectrum Disorder

Figure 2

ASD-NSCs display perturbations in neuronal phenotype during early days of differentiation. (a) Differentiated neurons from ASD-NSCs display shorter neurites when compared to neurons differentiated from control-NSCs. A strong staining of the cell body is seen in neurons derived from ASD-NSCs. Bar = 20 μm. and , , determined by an unpaired -test. (b) Immunostaining for other differentiation markers such as tuj1 or βIIItubulin and stem cell marker Mushashi1 (Mush1) also shows differential staining in neurons derived from ASD-NSCs than control-NSCs. Bar = 20 μm. and , , determined by an unpaired -test. (c) RNA was extracted from days 1, 3, 5, 7, 14, and 21, and qRT-PCR was conducted for miR-1290. A stable increase in expression was seen in control-NSCs during days of differentiation from day 1 to day 14 whereas at day 21 a significant downregulation in miR-1290 expression was observed in ASD-NSCs. An unpaired -test followed by the Holm-Sidak multiple correction post hoc test revealed significant differences in days 1 (), 3 (), and 5 (). Analysis on day 21 expression revealed a significance of (), determined by an unpaired -test. Data are represented as the . (d) Representative northern blot of RNA derived from day 21 differentiated from three individual donors from control- and ASD-NSCs; note that the anti-DIG signals for the mature form (∼21 nt) were only seen in the control cases. A prestained small molecular weight miRNA marker was used to monitor RNA size.

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