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Disease models represent powerful tools used by biomedical researchers to address various questions related to the pathogenesis and the possible therapeutic targets in different diseases. To get a complete picture of the process one needs complementary animal and ex vivo disease mo ...

Patients with pemphigus have circulating anti-desmoglein (Dsg) 1 and/or Dsg3 autoantibodies that induce blister formation on the skin and mucous membrane. We describe here two assays that measure the pathogenic strength of autoantibodies in blister formation: an in vitro dissocia ...

The characterization of the quality and quantity of cell death has gained substantial interest over the past decades. More recently necroptosis as a programmed form of necrosis has been identified as an important additional form of cell death with relevance in the skin. Understanding how to a ...

The terminal differentiation of epidermal keratinocytes requires transcriptional and posttranscriptional regulatory mechanisms. MicroRNAs (miRNAs) are small noncoding RNAs that play key roles during differentiation processes by regulating protein expression at ...

Physiological functions of skin cells are often altered in diseases. Since the underlying molecular mechanisms are generally executed by proteins, it is of interest to assess protein dynamics in normal and pathologically altered cells. These can be readily analyzed in relevant cell cu ...

Current knowledge of the morphological and molecular events driving branching morphogenesis of the ureteric bud (UB) during development of the metanephric kidney has been greatly facilitated by the ability to explant this organ to culture. The UB can be further isolated from the mesench ...

In vitro culture of embryonic kidney rudiments has been utilized to study a variety of cellular processes and developmental mechanisms. Here, we describe two-dimensional (2D) culture of embryonic kidney rudiments on Transwell filters and three-dimensional (3D) cultures in collag ...

Embryonic development is a highly dynamic process involving complex tissue interactions and movements. Recent progress in cell labeling, image acquisition, and image processing technologies has brought the study of embryo morphogenesis to another level. It is now possible to visu ...

The organ culture setup of embryonic kidney has served as a model of nephrogenesis for several decades. In vitro culture of the mouse metanephric mesenchyme enables easy manipulation and analysis of the tissue and provides information of cellular interactions, morphogenesis, cell di ...

Studies into the molecular basis of morphogenesis frequently begin with investigations into gene expression across time and cell type in that organ. One of the most anatomically informative approaches to such studies is the use of in situ hybridization, either of intact or histologically ...

This chapter describes detailed methods used for laser capture microdissection (LCM) of discrete subpopulations of cells. Topics covered include preparing tissue blocks, cryostat sectioning, processing slides, performing the LCM, and purification of RNA from LCM samples. Not ...

All the mRNAs within a cell and their relative levels are indicative of gene expression within that cell, which is essential for its structure and function in its differentiated state. Therefore, methods for the identification of the specific mRNAs and the quantitation of their levels are inv ...

The Genitourinary Development Molecular Atlas Project (GUDMAP) aims to document gene expression across time and space in the developing urogenital system of the mouse, and to provide access to a variety of relevant practical and educational resources. Data come from microarray gene ex ...

Renal development is a complex process involving the dynamic interplay of over 25 different cell types. One distinct step in this process is the formation of the ureteric tree, which develops from the repeated branching of the ureteric bud. During branching of the ureteric bud, cells migrate col ...

Immortalized cell lines derived from embryonic mouse kidneys are useful tools for exploring signaling pathways, morphogenetic mechanisms, and gene function in renal development: they also provide a means for efficient first-round screening of panels of small molecules intended ...

Primary cilia are present in most vertebrate cells. They have complex structures that are required for signal transduction in developing tissues. The embryonic kidney consists of two major cell lineages, ureteric and metanephric mesenchyme. Here, we describe a method to isolate metan ...

Kidney development has been widely used as a model system to study molecular control of inductive tissue interactions and mechanisms through which branching organs form. Due to lacking or poor methods, less focus has been in understanding details of cellular events that accomplish examp ...

This chapter presents three methods for re-constructing mouse foetal kidney tissue from simple suspensions of cells. These techniques are very useful for a number of purposes: (1) they allow the production of fine-grained chimaeras in which cell autonomy of mutations can be tested, (2) they p ...

Optical Projection Tomography (OPT) is an imaging technique, which has proven to be ideally suited to the observation and quantification of kidney development in rodents. Unlike confocal microscopy systems, OPT is capable of imaging the organ in toto across a long window of embryonic devel ...

Three-dimensional imaging is a valuable tool for analyzing kidney growth and development. This technique provides information about spatial relationships between the branching ureteric bud, nephrons, and other structures within the kidney. Availability of user-friendly v ...