Special technical expertise within this core includes extensive knowledge of skin and hair follicle morphology essential for understanding the genotype/phenotype correlations; state of the art methods for gene expression studies at both mRNA and protein level specifically designed for mouse and human skin; and possession of a battery of original experimental approaches to the study of skin physiology in vivo. The Core will assist Center investigators as well as other researchers at Columbia in evaluating the significance of a phenotypic manifestation or unanticipated skin effects of a genetic mutation, drug, or other experimental manipulation; to understand the possible functions of genes and molecular pathways implicated in skin/hair phenotype development and to analyze skin expression patterns of genes of investigator’s interest at the mRNA and protein level using immunohistochemistry and in situ hybridization. In addition, the Core will provide investigators with proof-reading of and suggestions for Materials & Methods sections for publications and applications for extramural research funding utilizing skin phenotype analysis. The Skin Phenotyping Core also serves as a center of communication between investigators providing the ground for exchange of samples, antibodies, techniques and ideas. The Core brings investigators working in similar projects and techniques together thus reducing the costs and increasing efficiency of research.
Analysis of skin/hair phenotype in mutant, genetically engineered, or drug-treated laboratory rodents using a set of relevant methodological approaches and techniques. This service includes: 1) gross (clinical) analysis of phenotype; 2) skin harvesting, preparation of frozen and paraffin blocks, sectioning and standard H&E histology; 3) a wide variety of special stains, which are available for localization of nerves, microvasculature, lipids, proteins, enzymes and some specific cell types (Langerhans cells, mast cells etc.) in skin. Currently, the most frequently used histochemistry techniques are oil red and alkaline phosphatase which are used for specific detection of lipid-producing cells (e.g., sebocytes) and
mesenchymal components of hair follicles (follicular papilla cells). Phenotype evaluation also includes skin/hair follicle morphometry, statistical analysis of obtained results, and photodocumentation using SPC imaging system.
Immunohistochemistry, immunofluorescence, and in situ hybridization in paraffin-embedded and frozen mouse/rat skin samples. The SPC has established standard protocols for skin immunohistochemistry and antigen retrieval which are extensively used for two major purposes: 1) to assess expression patterns of genes/proteins of interest in normal embryonic, newborn, and adult mouse skin at different stages of hair follicle morphogenesis and cycling; 2) to characterize the nature of defects in the skin of genetically engineered mice using antibodies for specific cytokines, transcription factors, as well as specific markers of proliferation, differentiation, apoptosis, and other cellular processes. The
SPC maintains a large library of specific antibodies, but if necessary, investigators can purchase primary and secondary antibodies in conjunction with the Core to reduce their cost, if another laboratory within the Core is interested in using the antibody.
Samples of normal mouse/rat skin are provided by the SPC which can be used by researchers for any kind of experimental studies (e.g., histology, histochemistry, immunohistochemistry, immunofluorescence, in situ hybridization) outside of the SPC. The SPC maintains a bank of frozen and paraffin embedded mouse and rat skin samples. The bank contains samples encompassing all stages of normal mouse and rat hair follicle morphogenesis
and cycling (both spontaneous and depilation-induced). The SPC will provide samples (blocks and sections) corresponding to any stage of normal hair follicle transformation necessary for expressional studies or as a wild type control for the studies of mutant or genetically engineered skin. SPC also maintains a bank of cDNA isolated from mouse whole skin at different stages of embryogenesis and hair follicle growth. These samples can be used for Northern blot analysis and qPCR.
CONTACT THE SKIN PHENOTYPING CORE
Dr. David Owens- firstname.lastname@example.org
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Last updated 03/30/2009