Growth alterations associated with IL34 genetic invalidations in zebrafish and mouse. (A) Scheme of IL34 Exon3 genetic alterations induced by CrispR/Cas9 technology in zebrafish. (B) Images of zebrafish mutants compared to the control at age of 3 months. (C) Mineralization of craniofacial skeleton by Von Kossa and Alcian Blue staining of embryos at 5 days post fecundation. Abbreviations: mx - branchio maxilla, bs - branchistegal ray, op - opercle, cl - cleithrum, pt - pharyngeal teeth, m - Meckel's cartilage, pq - palatoquadrate, ch - ceratohyal, ep - ethmoid plate, marked 1-5 - different arches. (D) Scheme of Il34 floxed allele used to obtain constitutive invalidation of IL34 in mouse by removing exons 3 to 5 under CRE recombinase activity. (E) Images at 15 days after birth of consequences of the constitutive invalidation of IL34 with detail of hydrocephaly in Il34-/- mouse (left panel). And comparative of skeletons at 15 postnatal days visualized by Alizarin red / Alcian blue double staining (right panel). (F) MicroCT scan 3D reconstructions of skull and tibia enable to visualize growth defects (red arrowheads). (G) Quantification of growth defects in the different morphometric planes (a to i) in wile type (black box) vs. Il34-/- mice (red box), both treated with IK22.5 RANKL blocking antibody (blue and green boxes), or wile type mice with Sheff.5 IL34 blocking antibody (brown box). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. The differences between the experimental conditions were assessed one-way ANOVA test. n=8 except for Il34-/- + IK22.5 (n=4).
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