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Fig. 4 | Neural Development

Fig. 4

From: Precise levels of nectin-3 are required for proper synapse formation in postnatal visual cortex

Fig. 4

Nectin-3 overexpression beginning at E15.5 decreases dendritic spine density. a Co-electroporated expression constructs. b Electroporation/nectin-3 overexpression occurred at E15.5. Dendritic spine densities were analyzed at P14, P21, and P35. c Western blot analysis after transfection of HEK-293 cells with a nectin-3 overexpression vector. Greatly increased nectin-3 expression is observed relative to endogenous levels in HEK-293 cells. d Representative image of an electroporated neuron with select apical and basal dendrites circled (scale bar = 50 μm). e Nectin-3 overexpression produced an overall decrease in dendritic spine densities compared to control (scramble shRNA) neurons (pooled apical and basal dendrites, solid line). As with control neurons, spine densities significantly increased between P14 and P21, indicating spinogenesis was intact with nectin-3 overexpression. Significance is denoted by ‘*’, and p values are listed in Additional file 5. f Representative images of apical dendrites from control and Nec3-OE neurons at P35 (63x image, scale bar = 10 μm). g Nec3-OE and control dendritic spine densities at P14. Apical and basal dendritic spine densities are plotted. Error bars denote standard error of the mean. Each dot represents a single dendrite. One apical and one basal dendrite per cell were counted (Control: N = 15 cells, 30 dendrites, 5 animals; Nec3-OE: N = 16 cells, 32 dendrites, 6 animals). h Nec3-OE and control dendritic spine densities were not significantly different at P21 (Control: N = 17 cells, 34 dendrites, 5 animals; Nec3-OE: N = 14 cells, 28 dendrites, 5 animals). When Nec3-OE and control data were combined, apical dendritic spine densities were significantly lower than basal at P21. i Dendritic spine densities for Nec3-OE neurons trended lower than control neurons at P35 (pooled apical and basal dendrites). This result was significant when considering apical dendrites alone (Control: N = 11 cells, 22 dendrites, 3 animals; Nec3-OE: N = 12 cells, 24 dendrites, 3 animals)

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