Figure 7: Role of Rbfox1-iso1 in the axon and dendrite growth in cortical neurons in vivo. | Scientific Reports

Figure 7: Role of Rbfox1-iso1 in the axon and dendrite growth in cortical neurons in vivo.

From: Essential role of the nuclear isoform of RBFOX1, a candidate gene for autism spectrum disorders, in the brain development

Figure 7

(a) Effects of Rbfox1-iso1-knockdown on the axon elongation. pCAG-RFP was coelectroporated with pSuper-H1.shLuc (Control) or pSuper-mRbfox1-iso1#1 into cerebral cortices at E14.5, and analyzed at P3. Hematoxylin staining of a slice was also shown (left panel). Scale bar, 1 mm. (b) Quantitative analyses of the ratio of the intensity of RFP-positive axons in the area (white) of the contralateral cortex to that in the area (green) of ipsilateral cortex in (a). Rescue experiments were done by cotransfection with pCAG-Myc-mRbfox1-iso1R. Error bars indicate SD; **p < 0.01 by Tukey-Kramer LSD (n = 3). (c) Representative images of the terminal arbors of callosal axons expressing RFP with pSuper-H1.shLuc (Control) or pSuper-mRbfox1-iso1#1 at P7. Scale bar, 200 μm. (d) Rbfox1-iso1-knockdown inhibits dendritic branching in vivo. pCAG-loxP-RFP was electroporated for sparse expression with pCAG-M-Cre together with pSuper-H1.shLuc (Control), pSuper-mRbfox1-iso1#1 or pSuper-mRbfox1-iso1#1 plus pCAG-Myc-mRbfox1-iso1R into cerebral cortices at E14.5. Analyses were carried out in cortical slices at P7. Representative average Z-stack projection images of RFP fluorescence of cortical neurons in upper CP were shown. Scale bar, 50 μm. (e,f) Number of dendritic branch points (e) or total dendritic length (f) was calculated on neurons observed in (d). Three brains were analyzed for each experiment; Control, n = 18 neurons; pSuper-mRbfox1-iso1#1, n = 22; pSuper-mRbfox1-iso1#1 plus pCAG-Myc-mRbfox1-iso1R, n = 21. Error bars indicate SD; **p < 0.01 by Tukey-Kramer LSD.

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