Kidney development : methods and protocols / edited by Odysse Michos. — New York : Humana Press : Springer, c2012. – (58.17/M592/v.886)

Contents

    Contents
    
    Preface
    Contributors
    PART I ORGAN CULTURE， TISSUE MANIPULATION， AND LIVE IMAGING
    1 Dissecting and Culturing and Imaging the Mouse Urogenital System
    2 In Vitro Culture of Embryonic Kidney Rudiments and Isolated Ureteric Buds 13
    3 In Vitro Induction of Nephrogenesis in Mouse Metanephric Mesenchyme with Lithium Introduction and Ureteric Bud Recombination 23
    4 Live Imaging of the Developing Mouse Mesonephros 31
    5 Organotypic Culture of the Urogenital Tract 45
    6 Live Imaging Kidney Development in Zebrafish 55
    PART II ANALYZING URETERIC BUD BRANCHING AND NEPHROGENESIS
    7 Analysis of 3D Branching Pattern: Hematoxylin and Eosin Method 73
    8 Three-Dimensional Imaging of Fetal Mouse Kidneys 87
    9 Analysis of Native Kidney Structures in Three Dimensions 95
    10 Estimating Nephron Number in the Developing Kidney Using the Physical Disector/Fractionator Combination 109
    11 An Immunofluorescence Method to Analyze the Proliferation Status of Individual Nephron Segments in the Xenopus Pronephric Kidney 121
    PART III RENAL PRIMARY CELL CULTURES AND CELL LINES
    12 Dissociation of Embryonic Kidney Followed by Re-aggregation as a Method for Chimeric Analysis 135
    13 Analysis of Migration in Primary Ureteric Bud Epithelial Cells 147
    14 Investigating Primary Cilia in Cultured Metanephric Mesenchymal Cells
    15 Making Immortalized Cell Lines from Embryonic Mouse Kidney
    16 Engineered Tissues to Quantify Collective Cell Migration During Morphogenesis 173
    PART IV DETECTING GENE/PROTEIN EXPRESSION AND SIGNALING
    17 Access and Use of the GUDMAP Database of Genitourinary Development 185
    18 Isolation of High Quality RNA from Embryonic Kidney and Cells 203
    19 Laser Capture 211
    20 Use of In Situ Hybridization to Examine Gene Expression in the Embryonic， Neonatal， and Adult Urogenital System 223
    21 Detection of β-Galactosidase Activity: X-gal Staining
    22 Fluorescent Immunolabeling of Embryonic Kidney Samples .
    23 Immunohistochemical Staining of dpERK Staining During Early Kidney Development 261
    24 Sensing BMP Pathway Activity by Immune Detection of Phosphorylated R-Smad Proteins in Mouse Embryonic Kidney 267
    25 Analysis of In Vivo Transcription Factor Recruitment by Chromatin Immunoprecipitation of Mouse Embryonic Kidney 275
    PART V MANIPULATING GENE EXPRESSION IN DEVELOPING AND ADULT KIDNEY
    26 siRNA-Mediated RNA Interference in Embryonic Kidney Organ Culture 295
    27 Morpholino-Mediated Gene Knockdown in Mammalian Organ Culture 305
    28 Microinjection into the Lumen of the Ureteric Tree 311
    29 Renal Delivery of Adenovirus and Antisense Oligonucleotides in Rats by Retrograde Renal Vein Injection 321
    PART VI ANALYZING FUNCTIONAL DEFECTS OF THE KIDNEY AND URINARY TRACT
    30 Estimating Total Nephron Number in the Adult Kidney Using the Physical Disector/Fractionator Combination 333
    31 Assessing Urinary Tract Defects in Mice: Methods to Detect the Presence of Vesicoureteric Reflux and Urinary Tract Obstruction 351
    32 Ischemia-Reperfusion Injury of the Mouse Kidney 363
    33 Variable Partial Unilateral Ureteral Obstruction and Its Release in the Neonatal and Adult Mouse 381
    34 Urinary Diversion via Cutaneous Vesicostomy in the Megabladder Mouse 393
    35 Ultrasound Imaging of the Murine Kidney 403
    Index 411