csf1r inhibitor blz945  (MedChemExpress)

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Gestational exposure to PLX5622 significantly depletes CSF1R-expressing cells. (A) Schematic illustrating collection of Csf1r EGFP + cells for flow cytometry. (B) Quantification of EGFP + cells from Csf1r EGFP craniofacial tissues ( n =3-10 embryos per sex/treatment/time-point from two to four dams). (C-W) Immunofluorescence images and quantification of CSF1R + and Csf1r EGFP+ cells in and around the E15.5 nasal septum (C-E), Meckel's cartilage (F-H), ear (I-K), maxillary incisor (L-N), eye (O-Q), trigeminal (R-T) and tongue (U-W). Arrows mark CSF1R and Csf1r EGFP double-positive cells ( n =3 embryos per sex/treatment from two or three dams). c.d., cochlear duct; d.p., dental papilla; e.k., enamel knot; ey, eye; l.s.c., lateral semicircular canal; m.c., Meckel's cartilage; n.s., nasal septum; s.r., stellate reticulum; tg, tongue; ut, utricle. Blue dots represent male and pink dots represent female. Counts represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques: Expressing, Flow Cytometry, Immunofluorescence

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Prenatal exposure to PLX5622 disrupts osteoclast development and function. (A-D′) Csf1r EGFP + osteoclasts (arrows) in the E15.5 premaxilla (A-B′) and maxilla (C-D′). (E-I) Immunofluorescence images (E-H) and quantification (I,I′) of multinucleated CTSK/ Csf1r EGFP double-positive osteoclasts (arrows) (I) and Csf1r EGFP+ single-positive osteoclasts (I′) in the E15.5 mandible. (J-S′) TRAP staining (arrows) in E15.5 premaxilla (J-K′), mandible (L-M′), maxilla (N-O′), frontal (P-Q′) and basioccipital (R-S′) bones. (T-V) Quantification of total TRAP + area in premaxilla (T), mandible (U) and maxilla (V). n =3 embryos per sex/treatment from two or three dams. bo, basioccipital; fr, frontal; m.c, Meckel's cartilage; md, mandible; mx, maxilla pmx, premaxilla. Counts represent mean±s.e.m. and were analyzed by an ART ANOVA (I,I′) or a two-way ANOVA (T-V) with Tukey's post-hoc test. A′-F′ show magnifications of the respective boxed areas in A-F.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques: Immunofluorescence, Staining

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Exposure to PLX5622 during embryogenesis increases bone density in the premaxilla and mandible. (A-J′) E15.5 Sp7 osteoblast staining in the premaxilla (A-B′) mandible (C-D′), maxilla (E-F′), frontal (G-H′) and basioccipital (I-J′) bones of CD1 embryos. (K-T′) E15.5 von Kossa and Alcian Blue staining in the premaxilla (K-L′), mandible (M-N′), maxilla (O-P′), frontal (Q-R′) and basioccipital (S-T′) bones of CD1 embryos. (U-W) Quantification of unmineralized area within the premaxilla (U) and mandible (V), and mineralized area in the maxilla (W). n =3 embryos per sex/treatment from two or three dams. bo, basioccipital; fr, frontal; m.c, Meckel's cartilage; md, mandible; mx, maxilla; pmx, premaxilla. Measurements represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques: Staining

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Prenatal exposure to PLX5622 disrupts craniofacial bone morphogenesis. (A-D′) Lateral (A-B′) and dorsal (C-D′) views of P1 CD1 skulls. Arrows in A-B′ indicate skull doming. Arrowheads in C-D′ mark interfrontal suture. Arrows in C-D′ mark sagittal suture. (E-G) Quantification of skull (E), interparietal (F) and occipital (G) bone lengths. (H-K) Lateral view of P1 CD1 mandibles (H-I′) and quantification of mandible length (J) and height (K). Arrows in H-I′ indicate measured lengths. (L-Q) Lateral view of the P1 CD1 ear (L-M′) and quantification of tympanic ring length (N) and thickness (O), and malleus (P) and stapes (Q) lengths. Arrows mark incus (absent in PLX5622 pups, asterisks), stapes and malleus. Arrowheads in L-M′ mark tympanic ring. Triangular arrowheads in L-M′ mark otic capsule. (R-U′) Ventral views of the P1 CD1 palate. Arrows in R-S″ mark palatine sutures. Arrowheads in R-S″ indicate altered palate morphology. (V) Quantification of premaxilla thickness. Double-headed arrows in R-S″ indicate measured thickness. (W-X′) Ventral view of the P1 CD1 cranial base. n =5-8 pups per sex/treatment from two to four dams. bo, basioccipital; bs, basisphenoid; fr, frontal bone; in, incus; ip, interparietal; ma, malleus; mx, maxilla; na, nasal bone; oc, occipital; pa, parietal bone; p.b., palatine bone; pmx, premaxilla; p.p.mx., palatine process of the maxilla; p.p.p., palatine process of the palatine; pt, pterygoid process; st, stapes; t.r., tympanic ring. Measurements represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques:

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Prenatal exposure to PLX5622 disrupts cranial base and brain development. (A-D) Lateral view of µCT scans of P1 CD1 skulls show presphenoid bone and inter-sphenoid synchondrosis. (E-K) Quantification of posterior cranial base length (E), anterior frontal complex length (F), basioccipital length (G), basisphenoid length (H), presphenoid length (I), spheno-occipital synchondrosis width (J) and inter-sphenoid synchondrosis width (K). (L-S) Coronal view of µCT scans of P1 CD1 skulls across the rostral (L-O) to caudal (P-S) axis. Arrows mark lateral (VL) and third (V3) ventricles. n =3-5 pups per sex/treatment from two or three dams. a.f.c., anterior frontal complex; bo, basioccipital; bs, basisphenoid; et, ethmoid; i.s.s., inter-sphenoid synchondrosis; p.c.b., posterior cranial base; ps, presphenoid bone; s.o.s., spheno-occipital synchondrosis. Measurements represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques:

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Exposure to PLX5622 during embryogenesis disrupts cytokine and chemokine secretion in craniofacial tissue cultures. (A) Schematic illustrating craniofacial tissue culture and Luminex multiplex analysis. (B-O) Cytokine/chemokine analysis of E13.5 craniofacial tissue culture media. n =9 per sex/treatment from three dams. Quantifications represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques: Luminex, Multiplex Assay

Journal: Development (Cambridge, England)

Article Title: CSF1R + macrophage and osteoclast depletion impairs neural crest proliferation and craniofacial morphogenesis

doi: 10.1242/dev.205423

Figure Lengend Snippet: Gestational exposure to PLX5622 decreases neural crest proliferation. (A) Schematic illustrating sphere assay. (B-M) E12.5 Wnt1 Cre ; Rosa26 tdTomato neural crest-derived spheres express PDGFRα (B,C,H,I), SOX2 (D,E,J,K) and SOX10 (F,G,L,M). (N) Primary sphere quantification ( n =6-9 embryos per sex/treatment from two or three dams). (O-Z) Immunofluorescence images of E15.5 Wnt1 Cre -driven tdTomato + signal (O-P′,U-V′) and Ki67 staining (R-S′,X-Y′) in the premaxilla (O-T) and mandible (U-Z). Quantification of tdTomato + (Q,W) and Ki67 + (T,Z) cells. White lines mark the tissue regions used for quantifications ( n =3 embryos per sex/treatment from three dams). DIV, days in vitro ; m.c., Meckel's cartilage; pmx, premaxilla. Counts represent mean±s.e.m. and were analyzed by a two-way ANOVA with Tukey's post-hoc test.

Article Snippet: Depletion of CSF1R + cells was achieved by administering the Plexxikon CSF1R inhibitor PLX5622 (1200 PPM added to chow AIN-76A, Research Diets) to pregnant female mice starting at E3.5.

Techniques: Derivative Assay, Immunofluorescence, Staining, In Vitro