Endocrine Methods
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Table of contents
- Cover
- Contentsv
- Contributorsxvii
- Prefacexxi
- Chapter 1. Assay Techniques Available for the Toxicologist1
- Introduction1
- Colorimetric Assays3
- Bioassays3
- Radioreceptor Assays6
- Radioimmunoassay9
- Nonradiometric Assays32
- Fluorescence Assays44
- Chemiluminescent Assays51
- Other Assay Techniques55
- Appendix 1: Selected Hormone Marker Assays60
- Conclusion/Addendum68
- References69
- Chapter 2. Longitudinal Bone Growth: Experimental Methods81
- Principles of Longitudinal Bone Growth81
- Measurement of Longitudinal Bone Growth82
- Growth Plate Infusion85
- References87
- Chapter 3. Measurement of Steroid Receptor Binding to DNA89
- Introduction89
- Mobility Shift Analysis90
- DNase I Footprinting94
- Methylation Interference97
- References99
- Chapter 4. Hypophysical Grafts beneath the Renal Capsule: A Model to Study Endocrine Control Systems101
- History and Utility of Pituitary Grafts101
- Comparison of the Pituitary Allograft Model with Studies Using Pituitary Tissue in Situ or in Vitro104
- Techniques and Considerations for Grafting Pituitary Tissue beneath the Renal Capsule105
- Summary of the Status of the Host and the Allograft Tissue113
- References113
- Chapter 5. Prolactin Secretion from Single Cells: Analysis by Reverse Hemolytic Plaque Assays115
- Introduction115
- Establishment of a Prolactin Plaque Assay117
- Procedures117
- Advantages of Using RHPA Analysis125
- Application of Plaque Assays to Assess Toxic Influence126
- References128
- Chapter 6. Application of Fluorescence Techniques to Bone Biology131
- Introduction131
- Cell Cultures132
- Analyses in Cell Populations135
- Analyses in Single Cells142
- References150
- Chapter 7. Measurement of Thyroid Hormone and Related Molecules157
- Introduction157
- Laboratory Measurements to Assess Thyroid Function158
- Autoimmune Processes That Influence Thyroid Gland Function and Hormone Measurements: Serum Thyroid A173
- Measurement of Serum Thyroglobulin176
- Conclusions177
- References178
- Chapter 8. Uptake Measurements of Thyroid Hormones and Amino Acidds in Cultured Cells187
- Introduction187
- Amino Acid Transport Systems188
- Uptake Measurements in Cultured Cells191
- Plasma Membrane Vesicles Derived from Cultured Cells for Uptake Measurements196
- Conclusion199
- References200
- Chapter 9. Two-Dimensional Gel Electrophoresis of Proteins203
- Introduction203
- Sample Preparation204
- The First Dimension: The ISO System205
- Alternate First Dimension: The IPG System207
- The Second Dimension: The DALT System207
- Detection of Protein Spots in 2D Gels209
- Protein Blotting212
- Analysis of 2D Protein Profiles by Computer- Based Image Analysis Systems217
- Summary218
- References219
- Chapter 10. Effects of in Vivo Exposure to Glucocorticoids on Pituitary, Serum, and Messenger Ribonu221
- Introduction221
- Methodologies223
- Experimental Protocols228
- Results229
- Summary235
- References238
- Chapter 11. Glutamate Receptor Autoradiography and in Situ Hybridization239
- Introduction239
- Methods for Studying Glutamate Receptor Expression244
- The Glutamate Receptor Binding Assay249
- Identification of Glutamate Receptor Subtypes by In Situ Hybridization Analysis260
- References271
- Chapter 12. Chromatographic Methods for Analyzing β-Endorphin and Related Peptides281
- Introduction281
- Procedures285
- Conclusions295
- References296
- Chapter 13. Detection of Autoantibodies to the Thyrotropin Receptor299
- The Thyrotropin Receptor and Normal Thyroid Function299
- The Function of the Thyrotropin Receptor300
- The Thyrotropin Receptor and Autoimmune Thyroid Diseases301
- Methods to Detect Autoantibodies to the Thyrotropin Receptor303
- Future Directions for Detection of Thyrotropin Receptor Antibodies311
- References315
- Chapter 14. Measurement of Intracellular Glucocorticoid and Mineralocorticoid Receptors319
- Introduction319
- Measurement of Intracellular GR and MR327
- Autoregulation of Rat Colonic GR and MR340
- References350
- Chapter 15. Use of Transfection Techniques for Studying the Function of the D2 Dopamine Receptors355
- Introduction355
- Transfection of Clonal Pituitary Cell Lines357
- Conclusions366
- References368
- Chapter 16. Quantitation of Metallothionein371
- The Physiology and Toxicology of Metallothionein371
- Quantitation of Metallothionein377
- Analysis of Metallothionein mRNA381
- Use of Metallothionein Analysis in Endocrine Toxicology382
- Summary and Conclusions383
- References385
- Chapter 17. Primary Culture of Bovine and Human Adult Adrenocortical Cells393
- Introduction393
- Materials and Reagents394
- Cell Culture of Bovine Adrenocortical Cells396
- Cell Culture of Human Adrenocortical Cells401
- Diagnostic Evaluation of Adrenocortical Cell Cultures405
- Conclusions414
- References415
- Chapter 18. Assays for Vasopressin417
- Background417
- Methodology420
- Application of RIA for Vasopressin433
- References440
- Index447
- Color Plate Section460
Endocrine Methods contains descriptions of contemporary and cutting-edge methodologies in various areas of endocrinology, including receptor theory and immunologic techniques for endocrine research. The book presents step-by-step procedures easily available to study the endocrine system and includes experts in their respective fields as contributors. The book presents step-by-step procedures for many important areas of endocrine target organs.
Endocrine Methods serves as a valuable methodological resource for investigators using endocrine methods.
Includes comprehensive, yet rapid methodical procedures
Offers a wide spectrum of assays including both in vivo and in vitro systems important to the several areas of hormone research
Describes several techniques for studying receptors, examining osteoblast activity, and measuring parathyroid hormones
Encompasses a host of important research tools that can be utilized by the toxicologist and other biomedical scientists
Endocrine Methods serves as a valuable methodological resource for investigators using endocrine methods.
Includes comprehensive, yet rapid methodical procedures
Offers a wide spectrum of assays including both in vivo and in vitro systems important to the several areas of hormone research
Describes several techniques for studying receptors, examining osteoblast activity, and measuring parathyroid hormones
Encompasses a host of important research tools that can be utilized by the toxicologist and other biomedical scientists