Project description:Viable mouse dermal macrophage (CD45+F4/80+SiglecF-) subtypes were identified by antigen expression combination of Ly6C, FRb and CD163 and sorted from dorsal skin of 7 week-old C57BL/6 female mice by FACS. Samples were paired within individual mice (5 mice each). RNA from 500 cells were isolated and profile for transcriptome by sequencing.
Project description:We analyzed dermal macrophage repertoire derived from PDGFRa-lineage or non-PDGFRa-lineage origin in atopic dermatitis (AD) with single-cell RNA sequencing. PDGFRa-lineage tracing mice aged between 7- and 14-week old were used to label cells of PDGFRa-lineage with tdTomato. AD was induced by topic application of MC903 bidaily or tridaily to skin of female mice for six times. Inflamed skin was excised on day 14 and single cells were prepared. Target cells were sorted as single cells for sequencing. We identified multiple subtypes in both PDGFRa-lineage or non-PDGFRa-lineage macrophage and the proportion differed by PDGFRa-lineage origin in AD skin.
Project description:To decipher macrophage heterogeneity and kinetics during atopic dermatitis, dorsal skin of C57BL/6 female mice aged between 7-12 week-old received MC903 stimulation bi- or tri-daily. Dermal leukocyte-enriched single cells were prepared and analysed by single-cell RNA-seq in time course.
Project description:Triplicate samples of donor matched ex vivo generated macrophage subtypes M1, M2a, M2c compared against the M0 control analysed using TMT-based 11plex quantitative proteomics.
Project description:Triplicate samples of donor matched ex vivo generated macrophage subtypes M1, M2a, M2c compared against the M0 control analysed using TMT-based 11plex quantitative proteomics.
Project description:Pain, detected by nociceptors, is an integral part of injury, yet whether and how it can impact tissue physiology and recovery remain understudied. Here we applied chemogenetics in mice to locally activate dermal TRPV1 innervations in naïve skin and found it triggered new regenerative cycling by dormant hair follicles (HFs).This was preceded by rapid apoptosis of dermal macrophages, mediated by the neuropeptide calcitonin gene-related peptide (CGRP). TRPV1 activation also triggered a macrophage-dependent induction of Spp1-expressing dermal fibroblasts. The neuropeptide CGRP andSpp1were essential for the nociceptor-triggered hair growth. Finally, we show that epidermal abrasion injury induced Spp1-expressing dermal fibroblasts and hair growth via a TRPV1 neuron and CGRP dependent mechanism.Collectively, these data demonstrate a role for TRPV1 nociceptors in orchestrating a macrophage and fibroblast-supported mechanism to promote hair growth, and enabling the efficient restoration of this mechano- and thermo-protective barrier after wounding.
Project description:Pain, detected by nociceptors, is an integral part of injury, yet whether and how it can impact tissue physiology and recovery remain understudied. Here we applied chemogenetics in mice to locally activate dermal TRPV1 innervations in naïve skin and found it triggered new regenerative cycling by dormant hair follicles (HFs).This was preceded by rapid apoptosis of dermal macrophages, mediated by the neuropeptide calcitonin gene-related peptide (CGRP). TRPV1 activation also triggered a macrophage-dependent induction of Spp1-expressing dermal fibroblasts. The neuropeptide CGRP andSpp1were essential for the nociceptor-triggered hair growth. Finally, we show that epidermal abrasion injury induced Spp1-expressing dermal fibroblasts and hair growth via a TRPV1 neuron and CGRP dependent mechanism.Collectively, these data demonstrate a role for TRPV1 nociceptors in orchestrating a macrophage and fibroblast-supported mechanism to promote hair growth, and enabling the efficient restoration of this mechano- and thermo-protective barrier after wounding.
Project description:A time course of the macrophage response to Salmonella exposure analyzing the effects of input cell number as a control for single cell studies Mouse macrophages were exposed to Salmonella enterica for different lengths of time. Libraries were constructed using either approximately 500,00 macrophages lysed directly on a tissue culture dish (bulk) or using only 150 cells isolated using FACS (sorted). All libraries were constructed in duplicate (bulk) or triplicate (sorted). All replicates are biological replicates