Detection of in vitro protein–DNA interaction is one of many investigational analyses for transcription factor regulation of gene promoters. The electrophoretic mobility shift assay (EMSA) has proven widely popular in this respect by integrating individual techniques (prot ...
Protein–DNA interactions, such as those that are necessary for transcription, are critical in regulating cellular function and behavior. The identification of DNA sequences that interact with transcriptional regulatory proteins is an important step necessary to better under ...
DNA microarray technology is a powerful tool for genome-wide gene expression analysis of biological samples including skin and epidermal keratinocytes. Here we review the methodology for expression profiling analysis of skin tissue or purified keratinocytes from mice. We expla ...
Epithelial tissues exhibit optimal conditions for studying cellular differentiation since the differentiation status of a single cell can be determined by its distance to the basal membrane. For that reason Laser Capture Microdissection (LCM) may serve as a perfect tool to compare the c ...
Because of its accessibility, skin has been among the first organs analyzed using DNA microarrays; psoriasis, melanomas, carcinomas, chronic wound biopsies, and epidermal keratinocytes in culture have been intensely investigated. Skin has everything: stem cells, differentia ...
The mouse mammary gland is a unique organ since although the mammary gland primordium forms during embryogenesis, the majority of development occurs postnatally upon hormonal stimulation at puberty and full functionality (i.e. lactation) is not achieved until after parturition. S ...
Although recent molecular biology advances provide a very effective way in determining localized gene expression, the visualization of the expression by in situ hybridization of whole-mount embryos or paraffin-embedded tissue sections continues to be an excellent method to det ...
Mouse skin has been used extensively as a model system to study the development of cancer. Recent emphasis has been focused on elucidating the molecular mechanisms by which chemical carcinogens or tumor promoters cause the growth of cutaneous malignancies. In this regard, many transgenic ...
The skin as a surface organ is uniquely accessible for whole embryo/foetal analyses of developmental changes, such as gene induction, protein expression, formation of epidermal-derived appendages such as hair follicles and formation of the protective barrier. Such analyses have em ...
The functional and molecular characterization of mouse epidermal stem cells (SC) requires in many circumstances arduous technologies and elaboration of complicated transgenic mouse models. To avoid these difficulties, we have developed a simplified protocol to obtain prima ...
Three-dimensional (3D) organotypic models are increasingly being used to study aspects of epidermal organisation and cutaneous wound-healing events. These are largely dependent on laborious histological analysis and immunohistochemical approaches. Here we outline a me ...
The ability to specifically manipulate gene expression in vivo using mouse models has been one of the most important advances in understanding gene function over the last few decades. Methods used to control gene activities in the mouse include gene targeting and transgenic approaches. Wh ...
The epidermis is a complex tissue composed principally of differentiated keratinocytes that form a keratinized stratified squamous epithelium. The gap junction proteins, connexins (Cx), are differentially expressed throughout the stratified layers of the epidermis and the ...
Prolonged mechanical loading of soft tissues may result in degeneration of these tissues, resulting in formation of pressure ulcers. The risk assessment of individuals might be improved by including measurements of the tissue response to mechanical loading. Cytokines, which are rel ...
Situations in which epidermal mutant mice display early lethality after birth are rather frequent. This condition precludes any kind of analysis of adult or even newborn mice tissues. We propose the in vivo embryonic skin transplantation as an alternative to solve this problem. This method ...
Human skin equivalents (HSEs) are in vitro tissues in which a fully differentiated, stratified squamous epithelium is grown at an air–liquid interface on a Type I collagen gel harboring human dermal fibroblasts. HSEs now provide experimental human tissue models to study factors that dire ...
Progress in the identification of skin stem cells and the improvement of culture methods open the possibility to use stem cells in regenerative medicine. Based on their quiescent nature, the development of label retention assays allowed the localization of skin stem cells in the bulge region ...
This protocol describes a technically simple in vivo assay of long-term skin regeneration in human skin, providing a reliable method for epidermal tissue reconstitution using small numbers of several types of epithelial cells, from epithelial cell lines to primary epithelial stem ce ...
The hair follicle is dynamic, cycling between growth (anagen), regression (catagen), and resting (telogen) phases throughout life. We have demonstrated that nestin-expressing hair follicle stem cells give rise to follicle structures during early anagen or growth phase of the hair fol ...
The basal layer of human interfollicular epidermis is thought to contain a minor compartment of quiescent or slowly cycling epithelial stem cells. These primitive keratinocytes give rise to the progenitors, which are the proliferating keratinocytes and which can be defined as early to l ...