Fig. 5

Overexpression of Nec3^Δafadin at E15.5 restricts spine formation after eye opening. a Co-electroporated expression constructs. b Representative image of an electroporated neuron with select apical and basal dendrites circled (scale bar = 50 μm). c Electroporation/dominant negative inhibition of nectin-3/afadin binding (Nec3^Δafadin) occurred at E15.5. Dendritic spine densities were measured at P14, P21, or P35. d Nec3^Δafadin produced an overall decrease in dendritic spine densities compared to control neurons when all time points were considered (pooled apical and basal dendrites, solid line). In addition, dendritic spine densities did not significantly increase between P14 and P21. Significance is denoted by ‘*’, and p values are listed in Additional file 5. e Representative images of basal dendrites at P21 from Nec3^Δafadin and control neurons (63x image, scale bar = 10 μm). f Representative images of apical dendrites at P35 from Nec3^Δafadin and control neurons (63x image, scale bar = 10 μm). g Nec3^Δafadin and control dendritic spine densities at P14. Apical and basal dendritic spine densities are plotted along with error bars denoting the standard errors of the mean (Control: N = 15 cells, 30 dendrites, 5 animals; Nec3^Δafadin, N = 16 cells, 32 dendrites, 6 animals). h Nec3^Δafadin and control dendritic spine densities at P21. Overall dendritic spine densities trended lower at P21 in Nec3^Δafadin neurons compared to control with a p value barely above the Bonferroni corrected p value of 0.0055. When basal dendrites were considered alone, a significant decrease in dendritic spine densities was observed (Control: N = 17 cells, 34 dendrites, 5 animals; Nec3^Δafadin: N = 12 cells, 24 dendrites, 3 animals). i Nec3^Δafadin and control dendritic spine densities at P35. When apical dendrites were considered alone Nec3^Δafadin spine densities were significantly lower than control (Control: N = 11 cells, 22 dendrites, 3 animals; Nec3^Δafadin: N = 9 cells, 18 dendrites, 3 animals)