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. 2012 Jan 18;32(3):1043–1055. doi: 10.1523/JNEUROSCI.4405-11.2012

Figure 5.

Figure 5.

Cortactin functions downstream of CTTNBP2 in the regulation of dendritic spine density. A, Coimmunoprecipitation of cortactin and CTTNBP2 (BP2) mutants. Whole-cell extracts of COS cells transfected with cortactin and wild-type or mutant CTTNBP2 (PA1, P540A/P543A; PA2, P599A/P602A) were precipitated with CTTNBP2 antibody. Immunoblotting (IB) was then performed to assess the presence of cortactin and CTTNBP2 in the precipitates. The arrowhead indicates the position of cortactin (top) or CTTNBP2 (bottom). The asterisk indicates a nonspecific signal. B–D, Rat hippocampal neurons were transfected with control miRNA (Ctrl-miR) or CTTNBP2 miRNA (BP2-miR) along with the Myc-tagged CTTNBP2 silent mutant (BP2-resc), the PA1 mutant (BP2-PA1-resc), or cortactin at 12 DIV. Neurons were harvested for staining with GFP and Myc-tag antibodies at DIV 18. B, Cortactin and the CTTNBP2 silent mutant restore the spine density in CTTNBP2 knock-down neurons. GFP signals were used to outline dendrite and spine morphology. Scale bar, 2 μm. C, Quantification of the density of protrusions. Eighteen neurons and >50 dendrites were assayed for each group. D, The width and length of protrusions assessed from >400 spines in each group. Error bars indicate mean ± SEM. *p < 0.05; ***p < 0.005.