Journal: Development (Cambridge, England)

Article Title: Bone morphogenetic proteins specify the retinal pigment epithelium in the chick embryo.

doi: 10.1242/dev.02884

Figure Lengend Snippet: Fig. 2. Comparison of the Sox10, Mitf and Bmp expression patterns during the initial stages of chick eye development. (A) Schematic illustrating the location of the section in H, showing the nasal part of the optic vesicle at stage 10/11. (B) Bmp4 expression in the surface ectoderm overlying the optic vesicle following whole-mount in situ hybridisation (13 somites). The optic primordium is shown at stage 8 (6 somites; C,F,I,L,O), at stage 9 (7-9 somites; D,G,J,M,P) and stage 10 (10-12 somites; E,H,K,N,Q). (C) At stage 8, Sox10-positive neural crest cells are observed in the dorsal neural folds of the prosencephalon (arrows). (D) Sox10-expressing neural crest cells are detected in the dorsal-most region of the prosencephalon (arrows) and no transcripts are detected distally. (E) Sox10 expression is detected in neural crest cells overlying the dorsal part of the optic vesicle (arrows). (F) At stage 8, Mitf expression is not observed in the optic primordium (arrow). (G) Mitf expression is strongest in the distal part of the optic vesicle at stage 9 (arrows). The arrowheads indicate neural crest cells dorsally. (H) At stage 10, Mitf expression is observed in the presumptive RPE (dorsal optic vesicle) and in the distal part (arrows). See A for the location of this section; see also R,S,T. (I) Bmp7 expression is observed in the ectoderm overlying the optic primordium at stage 8 (arrows), and diffuse expression is detected in the neural folds (arrowheads). (J) Parallel section of the embryo shown in G. At stage 9, strong Bmp7 expression is observed in the overlying ectoderm (arrows). Bmp7 transcripts are also detected in the dorsal ectoderm that covers the neural crest cells (arrowheads). (K) Bmp7 expression in the ectoderm overlying the distal region of the optic vesicle at stage 10 (arrows). Transcripts are still observed in the ectoderm overlying the mesenchyme (arrowhead). (L) At stage 8, Bmp4 transcripts are detected in the overlying ectoderm (arrow) and in the neural folds (arrowheads). (M) Strong Bmp4 expression is detected in the ectoderm overlying the distal portion of the optic vesicle at stage 9 (arrows). Weak or no expression is observed in the dorsal-most ectoderm overlying the mesenchymal cells (arrowheads). (N) At stage 10, Bmp4 expression is still strong in the ectoderm overlying the distal portion of the optic vesicle (arrows), whereas weak expression is observed in the ectoderm overlying the surrounding mesenchyme (arrowhead). Note that Bmp4 expression appears to be stronger in the ectoderm overlying the dorsal portion of the optic vesicle. (O) At stage 8, Bmp5 expression is strong in the dorsal midline, the neural folds (arrowheads). Transcripts appear to be absent from the overlying ectoderm. (P) Bmp5 expression weakens in the dorsal midline at stage 9 (arrowhead). (Q) No Bmp5 transcripts are detected in the neuroepithelium of the chick optic vesicle and surrounding tissues (arrow) at stage 10. (R) Nasal region of the optic vesicle at stage 10. Mitf expression in the distal and dorsal part of the optic vesicle (arrowheads). (S) Higher magnification of the Mitf expression pattern in the more-temporal region of the optic vesicle shown in H. Mitf expression is detected in both the dorsal and distal region of the optic vesicle (arrowheads), although expression weakens ventrally (arrow). (T) In the most-temporal region of the optic vesicle, Mitf expression is downregulated in the disto-ventral region (arrow).

Article Snippet: A 2 l drop of recombinant mouse BMP5 or BMP4 (0.7 mg/ml or 1 mg/ml; R&D Systems) was placed in a Petri dish and about eight drops (10 l each) of distilled water were placed around it to keep it from evaporating.

Techniques: Comparison, Expressing, In Situ, Hybridization

Journal: Development (Cambridge, England)

Article Title: Bone morphogenetic proteins specify the retinal pigment epithelium in the chick embryo.

doi: 10.1242/dev.02884

Figure Lengend Snippet: Fig. 3. Effects of BMP and noggin application on the Wnt2b expression pattern during early stages of eye development. (A) Schematic of a stage 10/11 chick embryo showing the implantation site of the BMP5-soaked bead in E. (B) At stage 13, Wnt2b transcripts are restricted to the presumptive RPE (arrow). The arrowhead indicates Wnt2b expression in the ectoderm. (C) Wnt2b transcripts are detected in the RPE (arrows) and surface ectoderm (arrowhead) on the contralateral side of the BMP5-treated eye shown in E. (D) Wnt2b expression in the contralateral, untreated eye following implantation of noggin-expressing cells. Wnt2b transcripts are restricted to the RPE (arrowhead) and no transcripts are detected within the NR. The arrow shows Wnt2b expression within the ectoderm and anterior lens. (E) Following BMP5 application (asterisk), Wnt2b expression is also detected in the distal region of the optic vesicle, the presumptive NR (arrows). The arrowhead indicates Wnt2b expression in the ectoderm. (F) Parallel section of the noggin-treated eye shown in Fig. 5B,D. Wnt2b expression is downregulated in the entire outer optic cup (arrowheads). In the surface ectoderm (arrow) and anterior lens, Wnt2b expression is still detected.

Article Snippet: A 2 l drop of recombinant mouse BMP5 or BMP4 (0.7 mg/ml or 1 mg/ml; R&D Systems) was placed in a Petri dish and about eight drops (10 l each) of distilled water were placed around it to keep it from evaporating.

Techniques: Expressing

Journal: Development (Cambridge, England)

Article Title: Bone morphogenetic proteins specify the retinal pigment epithelium in the chick embryo.

doi: 10.1242/dev.02884

Figure Lengend Snippet: Fig. 4. Effects of BMP5 application on the distribution of genes expressed within the NR and RPE at optic cup stages. (A) Schematic illustrating the location of the BMP5-soaked bead following implantation at stage 10/11 as shown in E-I; B-D are PBS-soaked bead controls (B) In control embryos, Mitf expression is weakly detected in the RPE at stage 15 (arrowheads). (C) The RPE-specific marker MMP115 is restricted to the RPE at this stage (arrowheads). (D) Strong Rx expression is detected in the NR at this stage (arrowhead). (E) Following implantation of a BMP5-soaked bead (asterisk), optic cup formation is not observed and Mitf expression is detected in the distal optic vesicle (arrowheads). (F) Parallel section of the embryo shown in E and G. MMP115 expression is induced in the presumptive NR (arrowhead) and the optic stalk region following BMP5 exposure. (G) Implantation of a BMP5-soaked bead (asterisk) leads to downregulation of Rx expression in the presumptive NR. (H) BMP5 application downregulates Chx10 expression in the distal optic vesicle/cup, the presumptive NR (right, arrowheads). By contrast, Chx10 expression is strongly observed within the presumptive NR of the contralateral eye (left, arrow). (I) Parallel section of the embryo shown in H. MMP115 expression is induced by BMP5 in the presumptive NR (arrowheads), whereas in the contralateral, unoperated eye, MMP115 transcripts are absent from the NR (left, arrow). L, Lens.

Article Snippet: A 2 l drop of recombinant mouse BMP5 or BMP4 (0.7 mg/ml or 1 mg/ml; R&D Systems) was placed in a Petri dish and about eight drops (10 l each) of distilled water were placed around it to keep it from evaporating.

Techniques: Control, Expressing, Marker

Journal: Nature Communications

Article Title: Bone marrow-derived epithelial cells and hair follicle stem cells contribute to development of chronic cutaneous neoplasms

doi: 10.1038/s41467-018-07688-8

Figure Lengend Snippet: CD34 − , CD44 + BMCs express keratin after BMC/KC co-culture and BMP5 treatment. a , b All adherent BMCs are CD34-negative and CD44-positive. c , e A sub set of adherent BMCs are pan-keratin-immunoreactive (arrowheads) after 7days of BMC/KC co-culture (keratin-positive BMCs in white boxes are magnified and merged with phase image). d Pan-keratin-immunoreactive BMC (arrowhead) identified 10 days after BMP5 treatment. f K14-immunoreactive cells (arrowheads) 10 days after BMP5 treatment (white box area is magnified). g Histogram of number of keratin-expressing BMCs; BMCs without treatment, BMC/KC co-culture (pan-keratin-positive BMCs, gray bar) and BMP5 treatment (K14-positive BMCs, black bar), pan-keratin- and K14-immunoreactive BMCs are detected in KC co-cultured and BMP5-treated BMCs, but no keratin-positive cells are detected in treatment controls ( n = 3, 3 different culture groups, 3 male and 3 female mice in each group; P < 0.017 as determined by Student’s t -test, mean ± s.d.) h Q-RT-PCR results show the relative level of K14 expression detected from positive (primary KCs and Tg.AC, a KC cancer cell-line) and negative (Swiss mouse 3T3 cell-line and primary BMCs) control groups, and experimental (BMC/KC co-culture and BMP5 treatment) groups. The expression levels were normalized to GAPDH expression levels, and expression mean values were converted into fold-change gene expression. Final values were normalized with log 2 transformation. ( n = 3, P < 4.81 × 10E−12 as determined by Fisher’s ANOVA method, mean ± s.d.). *White scale bar, 50 µm; Red scale bar, 200 µm

Article Snippet: For BMP5 (R&D systems, Minneapolis, MN; Cat# 6176-BM/CF) treatment, mesenchymal stem cell medium was changed every three days for 10 days, and BMP5 (50 ng/ml) was added immediately after changing medium (Supplementary Figure ).

Techniques: Co-Culture Assay, Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Control, Gene Expression, Transformation Assay