Endocrine Methods (eBook)

Front Cover 1
Endocrine Methods 4
Copyright Page 5
Contents 6
Contributors 18
Preface 22
Chapter 1. Assay Techniques Available for the Toxicologist 24
Introduction 24
Colorimetric Assays 26
Bioassays 26
Radioreceptor Assays 29
Radioimmunoassay 32
Nonradiometric Assays 55
Fluorescence Assays 67
Chemiluminescent Assays 74
Other Assay Techniques 78
Appendix 1: Selected Hormone Marker Assays 83
Conclusion/Addendum 91
References 92
Chapter 2. Longitudinal Bone Growth: Experimental Methods 104
Principles of Longitudinal Bone Growth 104
Measurement of Longitudinal Bone Growth 105
Growth Plate Infusion 108
References 110
Chapter 3. Measurement of Steroid Receptor Binding to DNA 112
Introduction 112
Mobility Shift Analysis 113
DNase I Footprinting 117
Methylation Interference 120
References 122
Chapter 4. Hypophysical Grafts beneath the Renal Capsule: A Model to Study Endocrine Control Systems 124
History and Utility of Pituitary Grafts 124
Comparison of the Pituitary Allograft Model with Studies Using Pituitary Tissue in Situ or in Vitro 127
Techniques and Considerations for Grafting Pituitary Tissue beneath the Renal Capsule 128
Summary of the Status of the Host and the Allograft Tissue 136
References 136
Chapter 5. Prolactin Secretion from Single Cells: Analysis by Reverse Hemolytic Plaque Assays 138
Introduction 138
Establishment of a Prolactin Plaque Assay 140
Procedures 140
Advantages of Using RHPA Analysis 148
Application of Plaque Assays to Assess Toxic Influence 149
References 151
Chapter 6. Application of Fluorescence Techniques to Bone Biology 154
Introduction 154
Cell Cultures 155
Analyses in Cell Populations 158
Analyses in Single Cells 165
References 173
Chapter 7. Measurement of Thyroid Hormone and Related Molecules 180
Introduction 180
Laboratory Measurements to Assess Thyroid Function 181
Autoimmune Processes That Influence Thyroid Gland Function and Hormone Measurements: Serum Thyroid Autoantibodies 196
Measurement of Serum Thyroglobulin 199
Conclusions 200
References 201
Chapter 8. Uptake Measurements of Thyroid Hormones and Amino Acidds in Cultured Cells 210
Introduction 210
Amino Acid Transport Systems 211
Uptake Measurements in Cultured Cells 214
Plasma Membrane Vesicles Derived from Cultured Cells for Uptake Measurements 219
Conclusion 222
References 223
Chapter 9. Two-Dimensional Gel Electrophoresis of Proteins 226
Introduction 226
Sample Preparation 227
The First Dimension: The ISO System 228
Alternate First Dimension: The IPG System 230
The Second Dimension: The DALT System 230
Detection of Protein Spots in 2D Gels 232
Protein Blotting 235
Analysis of 2D Protein Profiles by Computer- Based Image Analysis Systems 240
Summary 241
References 242
Chapter 10. Effects of in Vivo Exposure to Glucocorticoids on Pituitary, Serum, and Messenger Ribonucleic Acid Levels of the Gonadotropin Hormones FSH and LH in Male and Female Rats 244
Introduction 244
Methodologies 246
Experimental Protocols 251
Results 252
Summary 258
References 261
Chapter 11. Glutamate Receptor Autoradiography and in Situ Hybridization 262
Introduction 262
Methods for Studying Glutamate Receptor Expression 267
The Glutamate Receptor Binding Assay 272
Identification of Glutamate Receptor Subtypes by In Situ Hybridization Analysis 283
References 294
Chapter 12. Chromatographic Methods for Analyzing ß-Endorphin and Related Peptides 304
Introduction 304
Procedures 308
Conclusions 318
References 319
Chapter 13. Detection of Autoantibodies to the Thyrotropin Receptor 322
The Thyrotropin Receptor and Normal Thyroid Function 322
The Function of the Thyrotropin Receptor 323
The Thyrotropin Receptor and Autoimmune Thyroid Diseases 324
Methods to Detect Autoantibodies to the Thyrotropin Receptor 326
Future Directions for Detection of Thyrotropin Receptor Antibodies 334
References 338
Chapter 14. Measurement of Intracellular Glucocorticoid and Mineralocorticoid Receptors 342
Introduction 342
Measurement of Intracellular GR and MR 350
Autoregulation of Rat Colonic GR and MR 363
References 373
Chapter 15. Use of Transfection Techniques for Studying the Function of the D2 Dopamine Receptors 378
Introduction 378
Transfection of Clonal Pituitary Cell Lines 380
Conclusions 389
References 391
Chapter 16. Quantitation of Metallothionein 394
The Physiology and Toxicology of Metallothionein 394
Quantitation of Metallothionein 400
Analysis of Metallothionein mRNA 404
Use of Metallothionein Analysis in Endocrine Toxicology 405
Summary and Conclusions 406
References 408
Chapter 17. Primary Culture of Bovine and Human Adult Adrenocortical Cells 416
Introduction 416
Materials and Reagents 417
Cell Culture of Bovine Adrenocortical Cells 419
Cell Culture of Human Adrenocortical Cells 424
Diagnostic Evaluation of Adrenocortical Cell Cultures 428
Conclusions 437
References 438
Chapter 18. Assays for Vasopressin 440
Background 440
Methodology 443
Application of RIA for Vasopressin 456
References 463
Index 470
Color Plate Section 482