Human/Mouse FGF‑8 Antibody

Detection of Human/Mouse FGF-8 by Immunohistochemistry

Brefeldin A (BFA) treatment inhibits ERK1/2 activity & modulates differentially Fgf8 negative feed-back regulators. A, B & C) are 12 mM cryostat transversal sections of mouse ONTCs to the isthmic constriction. A & B) are control example before BFA administration to the culture medium showing an ISH for Fgf8 (A) & an Immunostaining for anti-FGF8 (B). Note different domains of expression of the transcript (solid line & arrows) & of the protein FGF8 (red arrows). FGF8 immunodetection is detected both at basal & ventricular sides of the ONTCs (black arrows in B). After 4 hours of BFA incubation (C-J) mRNA of Fgf8 was maintained at IsO (D) while the FGF8 protein profile changed dramatically being accumulated only at the ventricular side (D) as small vesicle-like, (arrows in fig. C & the magnified insert). Moreover, ERK1/2 activity disappears in Isthmic cells & nearby cells (E). Inside this negative gap, FGF8b beads still exerts polarizing ERK1/2 effects (F). Also inside this gap, genes such as Mkp3 (G) & Sef (H) disappear in mesencephalon while Sprouty family genes are maintained (I,J). K) represents experiments & model by which FGF8 planar induction activity coming from mouse FGF8-related secondary organizers (IsO & anr) exerts a different tissue preferential signaling effects (based on the activation of ERK1/2). The direction of polarized ERK1/2 activity depends on location of FGF8-related secondary organizers & the establishment of positional information signaling is dependent mainly on FGF8 negative modulator system, particularly Sprouty2 (blue gradient). FGF8 morphogenetic planar instruction signals coming from rostral (anr) & caudal (IsO) diminish & lose their polarization effect at the diencephalic region (zli) resulting in an equilibrium state. Scale bars is 100 mm. Image collected & cropped by CiteAb from the following publication (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3391221/), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Human/Mouse FGF-8 Antibody by Immunohistochemistry

Phosphorylation patterns of ERK1/2 (dpERK) enzymes in the anterior neural tube.Whole mount in situ hybridization (ISH) of E9.5 mouse embryo (A,B,B’) and corresponding organotypic neural tissue cultures of mouse E9.5 anterior neural tube (A’,B”,C-E”; ONTCs; [42]) where it shows the maintenance of gene expression profiles such us Fgf8, Tcf4 (A,A’) (used to delimit main brain subdivisions such the diencephalon (D; Tcf4 positive) and the mesencephalic anlage (M; negative staining), Pax6 D-D” (delimiting di-mesencephalic boundary and rhombomere 1–2 limits) and En2 E-E”. In B-B”) are photomicrographs of E9.5 mouse embryo with anti-dpERK Immunohistochemistry (IHC) taken from lateral (B) and caudal (B’) sides and the corresponding IHC in ONTCs. (C) double staining procedure: ISH (in blue) for Fgf8 and IHC for dpERK (dark brown) to localize inside the dpERK domain the position of the IsO, marked by the solid red line. (D-E”) photomicrographs of same ONTCs in which first a whole mount ISH for Pax6 (D) or Tcf4/En2 (E) were made and afterwards IHC against dpERK. (D’,E’ respectively). Dashed lines mark the main transversal (in black) and longitudinal (in red) brain subdivisions. These ONTCs were cut into transversal sections to the isthmic constriction (D”,E”) to proof that indeed dpERK expression reaches diencephalic anlage (D”; see asterisk; rostral is left) and has a wider expansion than En2 expression (E”; see asterisk; rostral is left). anr is anterior neural ridge secondary organizer; ba is branchial arch; IsO is isthmic organizer; os is optic stalk; ov is otic vesicle; r is rhombomere; T is telencephalon, D, diencephalon, M, mesencephalon. Scale bars are 0,5 mm except in D”, E” they are 100 µm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22792203), licensed under a CC-BY license. Not internally tested by R&D Systems.