Spermatogenesis: Methods and Protocols

ISBN-13: 9781627030373 / Angielski / Twarda / 2012 / 554 str.

Douglas T. Carrell; Kenneth I. Aston

Opis

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.

Spis treści

Section I. Semen Analysis Methods

 

1. Methods For Sperm Concentration Determination

            Lars Björndahl

 

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®)

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 Martínez-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. Montaño

 

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

Blurb

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™ 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.