Figure 5: ADAM10/17 double-knockout proprioceptive axons exhibit increased levels of Nrp1 at their growth cones. | Nature Communications

Figure 5: ADAM10/17 double-knockout proprioceptive axons exhibit increased levels of Nrp1 at their growth cones.

From: ADAM metalloproteases promote a developmental switch in responsiveness to the axonal repellant Sema3A

Figure 5

DRGs were dissected from E11.5 (a–c) and E12.5 (d–h) mice embryos of four different genotypes—wild type, ADAM10 cKO, ADAM17 cKO and ADAM10/17 double cKO. Explants were cultured in the presence of NT3 to facilitate growth of proprioceptive axons. Wild-type DRGs were cultured either with (b,e) or without (a,d) TAPI-1 (20 μM). Cultures of all four genotypes without TAPI-1 were supplemented with equivalent volumes of DMSO (carrier). Following fixation, the samples were surface stained (without permeabilization) with antibodies against Nrp1. (i) Relative fluorescence of growth cones from the different genotypes was measured. Results were normalized relative to average fluorescence of E11.5 wild-type growth cones with DMSO, which was set to 1; n (number of growth cones quantified) >1,200 for each condition. Results for each condition were collected from 3 to 7 separate experiments. (j) Growth cone fluorescence of mutant or pharmacologically inhibited neurons was compared with that of wild-type growth cones of the same embryonic day. Bars represent mean±s.e.m. Mann–Whitney U-test. ***P<0.001, NS, non significant. Scale bars, 25 μm. cKO, conditional knockout.

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