Kidney Development -

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Résumé :
Over the last decade the development of new molecular biology tools, advanced microscopy, live imaging and systems biology approaches have revolutionized our conception of how embryonic development proceeds.? One fundamental aspect of development biology is the concept of morphogenesis: understanding how a group of multipotent cells organize and differentiate into a complex organ.? In Kidney Development: Methods and Protocols, expert researchers in the field detail different approaches to tackle kidney development. These approaches include culture and live imaging aspects of kidney development, analyzing the 3-dimensional aspects of branching morphogenesis as well as nephrogenesis, manipulation of the gene/protein expression during kidney development as well as in the adult kidney, and how to assess kidney malformation and disease. Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. ? Authoritative and practical, Kidney Development: Methods and Protocols seeks to aid scientists in the further study of the process of morphogenesis which is fundamental important not only for studying developmental biology but also for regenerative medicine.

Sommaire:

?Dissecting and Culturing and Imaging the Mouse Urogenital System.-In Vitro Culture of Embryonic Kidney Rudiments and Isolated Ureteric Buds.-In vitro Induction of Nephrogenesis in Mouse Metanephric Mesenchyme with Lithium Introduction and Ureteric Bud Recombination.-Live Imaging of the Developing Mouse Mesonephros.-Organotypic Culture of the Urogenital Tract.-Live Imaging Kidney Development in Zebrafish.-Analysis of 3D Branching Pattern: Hematoxylin and Eosin Method.-Three-Dimensional Imaging of Fetal Mouse Kidneys.-Analysis of Native Kidney Structures in Three Dimensions.-Estimating Nephron Number in the Developing Kidney using the Physical Disector/Fractionator Combination.-An Immunofluorescence Method to Analyze the Proliferation Status of Individual Nephron Segments in the Xenopus Pronephric Kidney.-Dissociation of Embryonic Kidney Followed by Re-aggregation as a Method for Chimeric Analysis.-Analysis of Migration in Primary Ureteric Bud Epithelial Cells.-Investigating Primary Cilia in Cultured Metanephric Mesenchymal Cells.-Making Immortalized Cell Lines from Embryonic Mouse Kidney.-Engineered Tissues to Quantify Collective Cell Migration During Morphogenesis.-Access and use of the GUDMAP Database of Genitourinary Development.- Isolation of High Quality RNA from Embryonic Kidney and Cells.-Laser Capture.-Use of in Situ Hybridization to Examine Gene Expression in the Embryonic, Neonatal and Adult Urogenital System.-Detection of ?-galactosidase Activity: X-gal Staining.-Fluorescent Immunolabeling of Embryonic Kidney Samples.-Immunohistochemical Staining of dpERK Staining During Early Kidney Development.-SensingBMP Pathway Activity by Immune Detection of Phosphorylated R-Smad Proteins in Mouse Embryonic Kidney.-Analysis of in vivo Transcription Factor Recruitment by Chromatin Immunoprecipitation (ChIP) of Mouse Embryonic Kidney.-siRNA-mediated RNA Interference in Embryonic Kidney Organ Culture.-Morpholino-mediated Gene Knockdown in Mammalian Organ Culture.-Microinjection into the Lumen of the Ureteric Tree.-Renal Delivery of Adenovirus and Antisense Oligonucleotides in Rats by Retrograde Renal Vein Injection.-Estimating Total Nephron Number in the Adult Kidney using the Physical Dissector/Fractionator Combination.-Assessing Urinary Tract Defects in Mice: Methods to Detect the Presence of Vesico-ureteric Reflux and Urinary Tract Obstruction.-Ischemia-reperfusion Injury of the Mouse Kidney.-Variable Partial Unilateral Ureteral Obstruction and its Release in the Neonatal and Adult Mouse.-Urinary Diversion Via Cutaneous Vesicostomy in the Megabladder Mouse.-Ultrasound Imaging of the Murine Kidney.

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