mouse recombinant bmp5 protein  (Novus Biologicals)

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Four and a half domain 2 (FHL2) scaffolding protein is a marker of connective tissues of developing digits and regulates fibrogenic differentiation of limb mesodermal progenitors.

doi: 10.1002/term.2637

Figure Lengend Snippet: FIGURE 5 (a) Regulation of Fhl2 gene expression in micromass cultures 6 h after treatments with Wnt3A and Wnt5A (a), Tgfbeta2, and inhibitor of Smad3, SIS3 (b), and all‐trans‐retinoic acid (c). (d, e) Regulation of Fhl2 in micromass cultures 24 hr after treatment with different bone morphogenetic protein (BMP) ligands (BMP4, BMP5, BMP7, and GDF5; d) and BMP antagonists (noggin and chordin‐like 2) or the BMP inhibitor DMH1 (e). ***p < .001; **p < .01; *p < .05 versus control. ###p < .001 between the two treatments

Article Snippet: Fhl2 gene regulation was investigated after the following treatments: human recombinant TGFβ2 (10 ng/ml; R&D Systems); the specific inhibitor of Smad3, SIS3 (10 μM, Calbiochem); human recombinant BMP4, BMP5, BMP7, and GDF5 all at 200 ng/ml (R&D Systems); human recombinant NOGGIN at 200 ng/ml (R&D Systems); mouse recombinant chordin‐like 2 at 1200 ng/ml (R&D Systems); BMP inhibitor II, DMH1 at 4 mM (Calbiochem); human recombinant Wnt3a and Wnt5a at 150 ng/ml (R&D Systems); and all‐trans‐retinoic acid (RA) at 50 ng/ml (Sigma).

Techniques: Gene Expression, Control

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: Developmental biology

Article Title: A new role for BMP5 during limb development acting through the synergic activation of Smad and MAPK pathways.

doi: 10.1016/j.ydbio.2004.04.015

Figure Lengend Snippet: Fig. 2. (A–H) Expression of Bmp5 in the developing leg bud at days 3.5 (A), 4 (B), 5 (C, dorsal view; D, lateral view), 6 (E), 7.5 (F), 8 (G) and 9 (H). At early stages, transcripts are observed in the undifferentiated distal mesenchyme (A, B). Next in development in addition of being expressed in the distal mesoderm, transcripts are also observed in the dorsal and ventral muscle masses (arrows in C and D). As digits become identifiable, well-defined domains are progressively observed in the interdigital regions (E–G) which become displaced towards the tip of the digits in coincidence with the elimination of the most distal mesenchyme of the limb (H). Note the presence of additional domains of expression in the developing tendons (F, G) and in the perichondrium of the differentiating phalanxes (arrows, H).

Article Snippet: The function of BMP5 in the control of cell death and in chondrogenesis was studied by analyzing the effects of local administration into the limb mesoderm of recombinant BMP5 (R&D Systems) at concentrations ranging from 10 Ag/ml using as carriers heparin acrylic beads (Sigma).

Techniques: Expressing

Journal: Developmental biology

Article Title: A new role for BMP5 during limb development acting through the synergic activation of Smad and MAPK pathways.

doi: 10.1016/j.ydbio.2004.04.015

Figure Lengend Snippet: Fig. 3. Confocal images of squashed interdigital mesodermal cells showing the expression of bmp5 transcripts at day 6.5 of incubation (A). (B) Immunolabeling for BMP5 (red) combined with TUNEL (green) at day 7 of incubation. Note the intense cytoplasmic labeling both in living and in TUNEL- positive cells. (C–E) Apoptotic mesodermal cell after TUNEL and BMP5 immunolabeling. D and E show independent channels for BMP5 and TUNEL, respectively. (F, G) Double immunolabeling for ubiquitin (red) and BMP5 (green). Note the lack of co-localization between the two proteins both in the rounded apoptotic cell (F) and in the healthy stellate cell (G).

Article Snippet: The function of BMP5 in the control of cell death and in chondrogenesis was studied by analyzing the effects of local administration into the limb mesoderm of recombinant BMP5 (R&D Systems) at concentrations ranging from 10 Ag/ml using as carriers heparin acrylic beads (Sigma).

Techniques: Expressing, Incubation, Immunolabeling, TUNEL Assay, Labeling, Ubiquitin Proteomics

Journal: Developmental biology

Article Title: A new role for BMP5 during limb development acting through the synergic activation of Smad and MAPK pathways.

doi: 10.1016/j.ydbio.2004.04.015

Figure Lengend Snippet: Fig. 4. (A–I) Effects of local application of beads bearing BMP5 into the limb bud. (A, D) TUNEL-positive apoptosis induced by BMP5 beads (*) implanted in the anterior margin mesoderm of the wing bud at day 3.5 of incubation (A) and in the interdigital mesoderm before the onset of physiological apoptosis (D). (B, C) Experimental limbs vital stained with neutral red to show the area of cell death induced after implanting a BMP5 bead (*; B) and its inhibition by co- implantation of a Noggin bead (white *; C). E–F show similar results obtained when a BMP5 bead alone (E) or in combination with a Noggin bead (F) is implanted into the third interdigit. (G) Skeletal pattern of the wing bud lacking the radius (arrow) consequence of the experiment illustrated in B. (H, I) Neutral red staining of cell death (H) and skeletal staining with Alcian green (I) of limbs after implantation of a BMP5 bead at the tip of digit 3. Note that cell death is restricted to the most distal mesenchyme of the tip of the digit (H) while the digit cartilage exhibits intense overgrowth (*, I).

Article Snippet: The function of BMP5 in the control of cell death and in chondrogenesis was studied by analyzing the effects of local administration into the limb mesoderm of recombinant BMP5 (R&D Systems) at concentrations ranging from 10 Ag/ml using as carriers heparin acrylic beads (Sigma).

Techniques: TUNEL Assay, Incubation, Staining, Inhibition

Journal: Developmental biology

Article Title: A new role for BMP5 during limb development acting through the synergic activation of Smad and MAPK pathways.

doi: 10.1016/j.ydbio.2004.04.015

Figure Lengend Snippet: Fig. 5. (A–E) In situ hybridizations for BAMBI (A), Msx2 (B), Dkk (C), Snail (D) and Fgfr3 (E) 8 h after the application of a BMP5 bead in the third interdigit showing upregulation of all the genes. (F–J) Effect of beads soaked in SB203580 co-implanted with BMP5 beads in the expression of these genes. Note that BMP-mediated upregulation of Dkk (H), Snail (I) and Fgfr3 (J) is inhibited by the application of the p38 inhibitor SB203580, while the upregulation of BAMBI (F) and Msx2 (G) remains unchanged.

Article Snippet: The function of BMP5 in the control of cell death and in chondrogenesis was studied by analyzing the effects of local administration into the limb mesoderm of recombinant BMP5 (R&D Systems) at concentrations ranging from 10 Ag/ml using as carriers heparin acrylic beads (Sigma).

Techniques: In Situ, Expressing