Journal: Nature

Article Title: Distinct FGFs promote differentiation of excitatory and inhibitory synapses

doi: 10.1038/nature09041

Figure Lengend Snippet: a, fgf22 and fgf7 mRNAs are highly expressed in CA3 pyramidal neurons (arrowheads), but not in CA1 pyramidal neurons. Bottom panels are negative controls using FGFKO sections. b, fgfr2 mRNA is widely expressed throughout the hippocampus. c, FGF22 and FGF7 proteins are localized in CA3 synapse-rich areas. Pictured areas correspond to the boxed area in a. Scale bars, 500 μm (a, b) and 50 μm (c). SR: stratum radiatum, SL: stratum lucidum.

Article Snippet: Expression plasmids for FGF22 and FGF7 were generated by subcloning the full-length mouse fgf22 or fgf7 cDNA into the NheI-XhoI sites of APtag-5 (GenHunter).

Techniques:

Journal: Nature

Article Title: Distinct FGFs promote differentiation of excitatory and inhibitory synapses

doi: 10.1038/nature09041

Figure Lengend Snippet: a, VGLUT1 and VGAT clustering on FGF-transfected WT hippocampal neurons (labeled with GFP; 7 DIV). b, Defects in the clustering of VGLUT1 and VGAT on the dendrites of FGF22KO or FGF7KO CA3 pyramidal neurons (Py-positive; 14 DIV). The number (puncta/mm neurite) and size of puncta were quantified. c, The number (x1,000 puncta/mm2) and size of VGLUT1 and VGAT puncta on non-CA3 pyramidal neurons (Py-negative). d, The presynaptic defects in FGFKO cultures are rescued by expression of the corresponding FGF in postsynaptic CA3 pyramidal neurons. Data are normalized to WT. e, f, FGF22-EGFP localizes to glutamatergic, and FGF7-DsRed to GABAergic synapses (arrowheads). g, Endogenous FGF22 and FGF7 are colocalized with PSD95 or VGAT. h, FGF22-EGFP and FGF7-DsRed exhibit differential dendritic localization. i, j, The number (x1,000 puncta/mm2) and size of VGLUT1 (i) or VGAT (j) puncta after FGF bath application to WT cultures. k, VGLUT1 and VGAT staining intensity of control and FGFR2KO CA3 sections (P8). l, Normalized staining intensity of FGFR2KO sections. Error bars are s.e.m. Data are from 50–290 neurites or 8–66 fields from at least three experiments. Significant difference from control at *P < 0.05; **P < 0.01; ***P < 0.001, t-test (d, k, l) or ANOVA followed by Tukey test; #P=0.073 (i). Scale bars, 10 μm.

Article Snippet: Expression plasmids for FGF22 and FGF7 were generated by subcloning the full-length mouse fgf22 or fgf7 cDNA into the NheI-XhoI sites of APtag-5 (GenHunter).

Techniques: Transfection, Labeling, Expressing, Staining, Control

Journal: Nature

Article Title: Distinct FGFs promote differentiation of excitatory and inhibitory synapses

doi: 10.1038/nature09041

Figure Lengend Snippet: a–d, Representative whole-cell recordings of mEPSCs (a) and mIPSCs (c) from WT and FGFKO hippocampal neurons; frequency and amplitude of mEPSCs (b) and mIPSCs (d) in the indicated groups (18–24 neurons/group). e, Representative time-course of seizure development during kindling experiments. f, Percentage of mice kindled when about half of the control mice are kindled (~21 PTZ injections) from four independent experiments (4–6 mice/experiment). Error bars are s.e.m. Significant difference from control at *P < 0.05 and ***P < 0.001, ANOVA followed by Tukey test. g, Summary model. FGF22 and FGF7 from CA3 pyramidal neurons promote the differentiation of excitatory and inhibitory presynaptic terminals, respectively. FGF22 and FGF7 are localized at corresponding synapses and activate differential signaling pathways for specific presynaptic differentiation.

Article Snippet: Expression plasmids for FGF22 and FGF7 were generated by subcloning the full-length mouse fgf22 or fgf7 cDNA into the NheI-XhoI sites of APtag-5 (GenHunter).

Techniques: Control, Protein-Protein interactions

Journal:

Article Title: Role for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation

doi: 10.1073/pnas.062321799

Figure Lengend Snippet: In vitro effects of FGF7 on the development of rat embryo pancreatic epithelium. Pancreatic epithelia at E13.5 were dissected and cultured for 7 days with or without FGF7 (50 ng/ml). Before culturing and at the end of the culture period, the epithelia were photographed (A a–c) and sectioned. All sections were stained for endocrine markers (insulin + glucagon, in red) or acinar markers (carboxypeptidase-A, in green) (A d–f). (B) Quantification of epithelium volume before and after culturing with or without FGF7. (C) Quantification of the absolute volume occupied by the endocrine cells under the various conditions. **, P < 0.01. n, number of samples analyzed.

Article Snippet: Human recombinant FGF7 (R & D Systems) was diluted in PBS containing 0.1% BSA.

Techniques: In Vitro, Cell Culture, Staining

Journal:

Article Title: Role for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation

doi: 10.1073/pnas.062321799

Figure Lengend Snippet: FGF7 increased the absolute volume occupied by cells that stained negative for endocrine and acinar markers but positive for cytokeratin. (A) Dose-dependent effect of FGF7 on volume of the epithelium, absolute volume occupied by endocrine cells, and absolute surface area occupied by cells that stained negative for endocrine and acinar markers. (B) The cells that stained negative for insulin + glucagon (yellow), carboxypeptidase-A (in green), and vimentin (in blue) stained positive for cytokeratin (in red). These cells were present in increased numbers in epithelia grown with FGF7 at 50 ng/ml (b) as compared with epithelia grown without FGF7 (a) (see A for quantification). n, number of samples analyzed.

Article Snippet: Human recombinant FGF7 (R & D Systems) was diluted in PBS containing 0.1% BSA.

Techniques: Staining

Journal:

Article Title: Role for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation

doi: 10.1073/pnas.062321799

Figure Lengend Snippet: Cells that stained negative for endocrine and acinar markers differentiated into endocrine cells on FGF7 removal. (A) E13.5 rat epithelia were grown for 7 days with (d–f) or without (a–c) FGF7. Cultures were either stopped (a, d) or continued for 1 (b, e) or 2 (c, f) weeks without FGF7. The epithelia were sectioned and stained for insulin + glucagon (in red) and carboxypeptidase-A (in green). (B) Quantification of the absolute volume occupied by the endocrine cells. (C) Quantification of the absolute volume occupied by cells that stained negative for endocrine and acinar markers. Hatched bars, no FGF7 during the culture period. Solid bars, FGF7 was present during the first week of culture. *, P < 0.05; **, P < 0.01.

Article Snippet: Human recombinant FGF7 (R & D Systems) was diluted in PBS containing 0.1% BSA.

Techniques: Staining

Journal:

Article Title: Role for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation

doi: 10.1073/pnas.062321799

Figure Lengend Snippet: Endocrine cells differentiated from cells that proliferated in response to FGF7 exposure. E13.5 rat pancreatic epithelia were grown for 7 days with (b, d, f) or without (a, c, e) FGF7, then pulsed for 1 h with BrdUrd. Cultures were either stopped (a, b) or continued for 1 week without FGF7 (c–f). The epithelia were sectioned and stained for insulin + glucagon (in green) and BrdUrd (in red). e and f are enlargements of c and d, respectively. Arrowheads indicate cells that stained positive for both BrdUrd and endocrine hormones.

Article Snippet: Human recombinant FGF7 (R & D Systems) was diluted in PBS containing 0.1% BSA.

Techniques: Staining