Affiliation:
1. John Curtin School of Medical Research Australian National University Canberra Australia
2. Université Grenoble Alpes, CNRS UMR 5309, INSERM U1209 Institute for Advanced Biosciences Grenoble France
Abstract
AbstractHuman fertility is declining in Western countries, and it is becoming increasingly clear that male infertility plays a pivotal role in the overall fertility decline. To understand the process that drives successful male germ cell maturation, the study of spermatogenesis of model organisms, such as mice, is essential. Residual bodies (RBs) play an important role in the last stages of spermatogenesis. They are formed at the time when post‐meiotic spermatids undergo sequential differentiation steps so that the acrosome and flagellum are developed, the nucleus is markedly condensed, and the cytoplasm is lost. The masses of lost cytoplasm become RBs. Our recent work has shown that RB dynamics are highly sensitive to even small fertility defects. It was also noted that the transcriptome and proteome of RBs changes in response to spermatogenic defects. Thus, RBs represent an excellent and highly sensitive entity for studying male fertility. Previously published protocols for RB purification had some major limitations: they produced an RB fraction that was heavily contaminated with spermatozoa and erythrocytes or required tens of grams of starting material. In addition, most of the available protocols were developed for purification of RBs from rat testes. Here, we present a protocol that allows the isolation of 2.5‐3 × 106 RBs from mouse testes with a purity of 98% from only 1 g of starting material. The purified material can be used for various downstream applications to study male fertility, such as transcriptome and proteome analyses, super‐resolution microscopy, and electron and cryo‐electron microscopy, amongst many others. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol: An improved method for purification of the residual bodies from the seminiferous tubules of mice
Subject
Medical Laboratory Technology,Health Informatics,General Pharmacology, Toxicology and Pharmaceutics,General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Neuroscience
Reference14 articles.
1. Isolation of residual bodies and spermatid cytoplasm from the testis of the adult rat;An Der Lan‐Hochbrunn B. C.;Micron,1974
2. Apoptosis Is Physiologically Restricted to a Specialized Cytoplasmic Compartment in Rat Spermatids