Spermatogenesis
Description
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 BiologyT 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.
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Content
Methods For Sperm Concentration Determination.- Methods of Sperm Vitality Assessment.- The Hypo-osmotic Swelling Test for Evaluation of Sperm Membrane Integrity.- Sperm Morphology Classification: A Rational Method For Schemes Adopted By The World Health Organization.- Sperm Morphology Assessment Using Strict (Tygerberg) Criteria.- Methods For Direct And Indirect Antisperm Antibody Testing.- Manual Methods For Sperm Motility Assessment.- Computer-Aided Sperm Analysis (CASA) Of Sperm Motility And Hyperactivation.- The Hemizona Assay For Assessment Of Sperm Function.- The Sperm Penetration Assay for the Assessment of Fertilization Capacity.- Methods For The Assessment Of Sperm Capacitation And Acrosome Reaction Excluding The Sperm Penetration Assay.- Sperm DNA Fragmentation Analysis Using the TUNEL Assay.- Sperm DNA Damage Measured by Comet Assay.- Sperm Chromatin Structure Assay (SCSA®).- Sperm Aneuploidy Testing Using Fluorescence In Situ Hybridization (FISH).- Flow Cytometric Methods For Sperm Assessment.- Human Y Chromosome Microdeletion Analysis By PCR Multiplex Protocols Identifying Only Clinically Relevant AZF Microdeletions.- Sperm Cryopreservation Methods.- Density Gradient Separation Of Sperm For Artificial Insemination.- Recovery, Isolation, Identification and Preparation of Spermatozoa from Human Testis.- Enhancement of Sperm Motility Using Pentoxifylline and Platelet-Activating Factor.- Intracytoplasmic Morphology Selected Sperm Injection.- Sperm Selection For ICSI Using Annexin V.- Sperm Selection for ICSI Using the Hyaluronic Acid Binding Assay.- Sperm Selection Based On Electrostatic Charge.- Sex-Sorting Sperm Using Flow Cytometry/Cell Sorting.- Assessment of Spermatogenesis Through Staging of Seminiferous Tubules.- Immunohistochemical Approaches for the Study of Spermatogenesis.- Ultrastructural Analysis of Testicular Tissue and Semen by Transmission and Scanning Electron Microscopy.- Assessment Of Oxidative Stress In Sperm And Semen.-Improved Chemiluminescence Assay for Measuring Antioxidant Capacity of Seminal Plasma.- Methods of Sperm DNA Extraction for Genetic and Epigenetic Studies.- Isolating mRNA and Small Noncoding RNAs From Human Sperm.- A Review Of Genome-Wide Approaches To Study The Genetic Basis For Spermatogenic Defects.- Methods For The Analysis Of The Sperm Proteome.- Methodology of Aniline Blue Staining Of Chromatin and the Assessment of the Associated Nuclear and Cytoplasmic Attributes in Human Sperm.- Isolation of Sperm Nuclei and Nuclear Matrices from the Mouse, and Other Rodents.- Extraction and Analysis of Human Sperm Protamine 1 / Protamine 2 Ratio Using Acid Gel Electrophoresis.- Analysis of Gene-Specific and Genome-Wide Sperm DNA Methylation.- Evaluating the Localization and DNA Binding Complexity of Histones in Mature Sperm.- In Vitro Spermatogenesis Using An Organ Culture Technique.- Testicular Tissue Grafting and Male Germ Cell Transplantation.- Transgenic Modification of Spermatogonial Stem Cells Using Lentiviral Vectors.- Methods for Sperm-Mediated Gene Transfer.- Phenotypic Assessment of Male Fertility Status in Transgenic Animal Models.