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 ...

Accurate analysis of the three-dimensional (3D) architecture of developing organs is critical to understanding how developmental defects can be linked with structural abnormalities. Here, we describe a 3D reconstruction technique of the developing kidney including the outer k ...

Organ development requires the coordination of proliferation and differentiation of various cell types. This is particularly challenging in the kidney, where up to 26 different cell types with highly specialized functions are present. Moreover, even though the nephron initially d ...

Design-based stereology is considered the gold-standard method for estimating the total number of glomeruli, and thereby nephrons, in the adult kidney. However, until recently, a design-based method for estimating nephron number in the developing kidney was not available. For such a me ...

The zebrafish has emerged as a powerful model to study organ development and regeneration. It has a number of advantages over other vertebrate model systems. The embryo can be kept transparent throughout embryonic development, which allows direct visualization of the developing orga ...

Organotypic culture is an invaluable technique that allows researchers with the tool to analyze a tissue development in an isolated and well-defined environment. This technique also permits one to study the roles of different signaling systems/signaling molecules and to take advan ...

X-gal staining is a rapid and convenient histochemical technique used to detect reporter gene expression. A prerequisite is the creation or acquisition of transgenic reporter mouse lines, in which the bacterial LacZ gene has been knocked into the gene of interest or placed under the control of ...