CiNii Books - Spermatogenesis : methods and protocols

Deficiencies in sperm function are usually the result of spermatogenic defects. Spermatogenesis is a biologically complex and essential process during which spermatogonia undergo meiotic recombination, reduction of the genome to a haploid state, and extensive cellular modifications that result in a motile cell capable of traversing the female reproductive tract, withstanding various potential assaults to viability, and finally successfully fertilizing a mature oocyte to give rise to an embryo. Defects in any step of spermatogenesis or spermatogenesis can lead to male infertility, a disease that affects approximately 5-7% of the population. Spermiogenesis and Spermatogenesis: Methods and Protocols details protocols used in the study of spermatogenesis, clinical analytical protocols, and basic techniques used in clinical andrology laboratories, such as obtaining accurate results for a sperm count, and advanced procedures, such as genome-wide genetic study tools and evaluation of nuclear proteins. Written in the successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Spermiogenesis and Spermatogenesis: Methods and Protocols is unique in its breadth, and will be a useful reference for clinicians and researchers alike.

Table of Contents

Section I. Semen Analysis Methods 1. Methods For Sperm Concentration Determination Lars Bjoerndahl 2. Methods of Sperm Vitality Assessment Sergey I. Moskovtsev and Clifford L. Librach 3. The Hypo-osmotic Swelling Test for Evaluation of Sperm Membrane Integrity Sivakumar Ramu and Rajasingam S. Jeyendran 4. Sperm Morphology Classification: A Rational Method For Schemes Adopted By The World Health Organization Susan A. Rothmann, Anna-Marie Bort, John Quigley, and Robin Pillow 5. Sperm Morphology Assessment Using Strict (Tygerberg) Criteria Roelof Menkveld 6. Methods For Direct And Indirect Antisperm Antibody Testing Hiroaki Shibahara and Junko Koriyama 7. Manual Methods For Sperm Motility Assessment David Mortimer and Sharon T. Mortimer 8. Computer-Aided Sperm Analysis (CASA) Of Sperm Motility And Hyperactivation David Mortimer and Sharon T. Mortimer Section II. Tests of Sperm Function 9. The Hemizona Assay For Assessment Of Sperm Function Sergio Oehninger, Mahmood Morshedi, and Daniel Franken 10. The Sperm Penetration Assay for the Assessment of Fertilization Capacity Kathleen Hwang and Dolores J. Lamb 11. Methods For The Assessment Of Sperm Capacitation And Acrosome Reaction Excluding The Sperm Penetration Assay Christopher J. De Jonge and Christopher L. R. Barratt Section III. Sperm DNA Damage Testing 12. Sperm DNA Fragmentation Analysis Using the TUNEL Assay Rakesh Sharma, Jayson Masaki, and Ashok Agarwal 13. Sperm DNA Damage Measured by Comet Assay Luke Simon and Douglas T. Carrell 14. Sperm Chromatin Structure Assay (SCSA (R)) Donald P. Evenson Section IV. Advanced Clinical Testing 15. Sperm Aneuploidy Testing Using Fluorescence In Situ Hybridization (FISH) Benjamin R. Emery 16. Flow Cytometric Methods For Sperm Assessment Vanesa Robles and Felipe Martinez-Pastor 17. Human Y Chromosome Microdeletion Analysis By PCR Multiplex Protocols Identifying Only Clinically Relevant AZF Microdeletions Peter H. Vogt and Ulrike Bender Section V. Sperm Preparation and Selection Techniques 18. Sperm Cryopreservation Methods Tiffany Justice and Greg Christensen 19. Density Gradient Separation Of Sperm For Artificial Insemination David Mortimer and Sharon T. Mortimer 20. Recovery, Isolation, Identification and Preparation of Spermatozoa from Human Testis Charles H. Muller and Erin R. Pagel 21. Enhancement of Sperm Motility Using Pentoxifylline and Platelet-Activating Factor Shilo L. Archer and William E. Roudebush 22. Intracytoplasmic Morphology Selected Sperm Injection Luke Simon, Aaron Wilcox, and Douglas T. Carrell 23. Sperm Selection For ICSI Using Annexin V Sonja Grunewald and Uwe Paasch 24. Sperm Selection for ICSI Using the Hyaluronic Acid Binding Assay Mohammad Hossein Nasr-Esfahani and Tavalaee Marziyeh 25. Sperm Selection Based On Electrostatic Charge Luke Simon, Shao-qin Ge, and Douglas T. Carrell 26. Sex-Sorting Sperm Using Flow Cytometry/Cell Sorting Duane L. Garner, K. Michael Evans, and George E. Seidel, Jr. Section VI. Staging and Histology Techniques 27. Assessment of Spermatogenesis Through Staging of Seminiferous Tubules Marvin L. Meistrich and Rex A. Hess 28. Immunohistochemical Approaches for the Study of Spermatogenesis Cathryn A. Hogarth and Michael D. Griswold 29. Ultrastructural Analysis of Testicular Tissue and Semen by Transmission and Scanning Electron Microscopy Hector E. Chemes Section VII. Oxidative Stress Testing 30. Assessment Of Oxidative Stress In Sperm And Semen Anthony H. Kashou, Rakesh Sharma, and Ashok Agarwal 31. Improved Chemiluminescence Assay for Measuring Antioxidant Capacity of Seminal Plasma Charles H. Muller, Tiffany K. Y. Tang, and Michalina A. Montano Section VIII. Analytical Tools and Methods 32. Methods of Sperm DNA Extraction for Genetic and Epigenetic Studies Jeanine Griffin 33. Isolating mRNA and Small Noncoding RNAs From Human Sperm Robert J. Goodric, Ester Anton, and Stephen A. Krawetz 34. A Review Of Genome-Wide Approaches To Study The Genetic Basis For Spermatogenic Defects Kenneth I. Aston and Donald F. Conrad 35. Methods For The Analysis Of The Sperm Proteome Sara de Mateo, Josep Maria Estanyol, and Rafael Oliva Section IX. Methodologies to Evaluate Sperm Epigenetics 36. Methodology of Aniline Blue Staining Of Chromatin and the Assessment of the Associated Nuclear and Cytoplasmic Attributes in Human Sperm Leyla Sati and Gabor Huszar 37. Isolation of Sperm Nuclei and Nuclear Matrices from the Mouse, and Other Rodents W. Steven Ward 38. Extraction and Analysis of Human Sperm Protamine 1 / Protamine 2 Ratio Using Acid Gel Electrophoresis Lihua Liu, Kenneth I. Aston, and Douglas T. Carrell 39. Analysis of Gene-Specific and Genome-Wide Sperm DNA Methylation Saher Sue Hammoud, Bradley R. Cairns, and Douglas T. Carrell 40. Evaluating the Localization and DNA Binding Complexity of Histones in Mature Sperm David Miller and Agnieszka Paradowska Section X. In Vitro and Ex Vivo Spermatogenesis Models 41. In Vitro Spermatogenesis Using An Organ Culture Technique Tetsuhiro Yokonishi, Takuya Sato, Kumiko Katagiri, and Takehiko Ogawa 42. Testicular Tissue Grafting and Male Germ Cell Transplantation Jose R. Rodriguez-Sosa, Lin Tang, and Ina Dobrinski Section XI. Transgenic Techniques 43. Transgenic Modification of Spermatogonial Stem Cells Using Lentiviral Vectors Christina Tenenhaus Dann 44. Methods for Sperm-Mediated Gene Transfer Marialuisa Lavitrano, Roberto Giovannoni, and Maria Grazia Cerrito. 45. Phenotypic Assessment of Male Fertility Status in Transgenic Animal Models David M. de Kretser and Liza O'Donnell

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