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. 2019 Dec 23;8:e49819. doi: 10.7554/eLife.49819

Figure 1. Nervous system specific VEGFR2 loss affects dendritic development in CA3 pyramidal neurons.

(A, B) Immunostainings for VEGFR2 revealed high levels of the receptor in the vessels (arrowheads) and double-labeling with the neuronal marker NeuN shows that the receptor is also expressed in pyramidal neurons (arrows) of the CA3 region in P10 hippocampus. Scale bar: A: 500 µm, B: 100 µm. (C–G) Nervous system specific deletion of VEGFR2 results in reduced dendritic arborization of CA3 pyramidal neurons. Nes-cre Kdrlox/- mice were crossed to Thy1-GFP transgenic mice to visualize whole morphology of pyramidal neurons. Z-projections of confocal images and the corresponding outlined tracings from CA3 neurons of P10 Nes-cre Kdrlox/- and control littermates are shown in (C). Total dendritic length (D), number of branch points (E) and the number of dendrites at 200 µm from the soma (F) were significantly reduced in Nes-cre Kdrlox/- mice compared to control littermates. 3D Sholl analysis of confocal z-stacks shows reduced branching and dendritic complexity of Nes-cre Kdrlox/- CA3 pyramidal neurons (G). Scale bar: 100 µm. n = 3 mice per genotype; SEM; *p<0.05.

Figure 1.

Figure 1—figure supplement 1. Characterization of a conditional, nervous system specific VEGFR2 knockout mouse.

Figure 1—figure supplement 1.

(A) Kdr-GFP knockin mouse line shows GFP expression in the vessels and CA3 neurons in P8 hippocampus. Scale bar: 200 µm. (B) PCR analysis confirmed the efficient recombination of the floxed Kdr exon one in DNA extracted from Nes-cre Kdrlox/- cultured neurons. (C) Quantitative RT-PCR shows a nearly complete elimination of Kdr mRNA isolated from knockout neurons (n = 3 experiments). Data are represented as mean ± SEM. ***p<0.001. (D) Immunohistochemistry for VEGFR2 shows the absence of the signal in fibers of the fimbria of Nes-cre Kdrlox/- mice (arrows), whereas the vessels normally express the receptor (arrowheads). Scale bar: 100 µm.