cgrp 8 37  (Tocris)

Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: (A) Diagram of experimental design and representative images from shaved TRPV1 activated mice and controls. Mice monitored for hair coat growth for 12 days after the last TRPV1 activation with CNO (postnatal day 63). (B) Enlarged dashed rectangle from panel A depicting hair growth in a TRPV1 activated mouse. (C) HF length quantification of dorsal skin from mice treated as in A. Data combined from 6 to 8 HF per mouse, taken from 6 mice per group (****p<0.0001). (D) Representative immunofluorescent images of dorsal skin from mice treated as in A. Staining with anti-keratin14 (KRT14; green) marking epidermal layer and Ki67 (purple) marking the HF bulb. DAPI stain in blue. Scale bar 100μm. (E) Quantification of normalized Ki67 MFI signal in the peri-follicular area of dorsal skin from mice treated as in A. Data combined from 4 regions of interest (ROI) per mouse taken from 5 TRPV1 activated mice and 9 controls (****p<0.0001). (F) Diagram of experimental design and representative images from TRPV1 activated mice and controls treated with CNO on half of their back skin (marked with dashed rectangle). Anti-Spp1 neutralizing antibody or IgG control were injected intradermally to the same skin area. Data are combined from 4 HFs per mouse collected from 6 controls, 3 TRPV1 activated + IgG treated and 4 TRPV1 activated + anti-Spp1 treated mice (****p<0.0001). (G-H) Percentage of CD9+CD26+ dermal fibroblasts from TRPV1 activated mice and their controls intradermally injected with (G) QWF (SubP antagonist) or vehicle (**p=0.005, ***p=0.0004), (H) CGRP8–37 (CGRP antagonist) or vehicle (***p=0.0001, ****p<0.0001). (I) Diagram of experimental design and representative images from TRPV1 activated mice and controls pre-treated intradermally with CGRP8–37 or vehicle and monitored for hair coat growth for 14 days. Quantification of HF length from 6 to 8 HFs per mouse, taken from 3–6 mice per group (****p<0.0001). (J) Representative immunofluorescent images of dorsal skin from mice treated as in I. Stained with anti-KRT14 (green), Ki67 (pink), and DAPI (blue). Scale bar, 100μm.

Article Snippet: CGRP 8-37 (rat) , Tocris , Cat# 1169.

Techniques: Activation Assay, Staining, Control, Injection

Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: (A) Quantification of TUNEL+ F4/80+ cell number per field of view from dorsal skin of TRPV1 activated mice and controls 3.5 hours after a single intradermal CNO treatment. Data are combined from 6 to 7 fields of view per mouse, taken from 7 mice per group (****p<0.0001). (B) Representative immunofluorescent images of 20μm skin sections from mice treated as in A. Sections stained with anti-F4/80 (red), TUNEL (green) and DAPI (blue). White arrowheads indicate F4/80+ TUNEL+ cells. Scale bar 50μm. (C) Percentage of Annexin V+ dermal macrophages from dorsal skin of TRPV1 activated mice and controls 45 minutes after a single CNO intradermal injection (***p=0.0002). (D) Mice back skin intradermally injected with CGRP on one side and vehicle on the other. (Left) Samples collected 45 minutes later and stained for annexin V as in C (*p-paired=0.017). (Right) samples collected 6 hours later and dermal macrophage frequency assessed (*p-paired=0.015). (E) Representative immunofluorescent images of 150μm dorsal skin sections from TRPV1 activated mice and controls pretreated with CGRP8–37 or vehicle followed by single intradermal CNO injection. Samples collected 6 hours later and stained with anti-F4/80 (red), CD207 (green), and DAPI (blue). Dashed rectangles indicate the lower follicle and peri-follicular area. Scale bar 100μm. (F) Quantification of F4/80 normalized MFI signal in the peri-follicular area from mice treated as in E. Data are combined from 6–8 ROIs per mouse taken from 5–8 mice per group (****p<0.0001). (G) Percentage of dermal macrophages from TRPV1 activated mice and controls treated as in E (*p=0.048,0.012,****p<0.0001). (H) Dermal macrophages frequency in dorsal skin of TRPV1 activated mice and controls reconstituted with Ramp1 KO or wild type BM. Chimeric mice given a single CNO injection and data collected 6 hours later (*p=0.04,***p=0.0009). (I) Dermal macrophages frequency in dorsal skin of TRPV1 activated mice and controls reconstituted with CD64Cre Ramp1+/+ or CD64Cre Ramp1fl/fl BM. Mice treated with CNO as in H (*p=0.043,***p=0.0001).

Article Snippet: CGRP 8-37 (rat) , Tocris , Cat# 1169.

Techniques: TUNEL Assay, Staining, Injection

Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: Key resources table

Article Snippet: CGRP 8-37 (rat) , Tocris , Cat# 1169.

Techniques: Recombinant, RNAscope, Multiplex Assay, In Situ, Software