Drosophila Oogenesis

This detailed volume compiles numerous methods to explore fly oogenesis. Beginning with updated protocols from isolating and staining the ovary to numerous imaging techniques and genetic protocols for cell-specific assessment and CRISPR-mediated mutagenesis, the volume continues by detailing techniques from assessing cytoskeletal structures to uncovering protein-protein interactions, with closing chapters exploring how Drosophila oogenesis can be used in the classroom and in outreach programs to increase interest in biomedical research, STEM education, and STEM careers. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Drosophila Oogenesis: Methods and Protocols serves as an ideal guide for both new and expert researchers working with this robust and versatile model organism.

The genetic development of eggs in fruit fly (Drosophilia) is the most important model used by developmental biologists in understanding how we go from a fertilized egg to a fully developed organism. Insights into fruit fly genetics have also begun to provide an idea as to how humans develop. This text amasses and organizes information on nearly 400 genes affecting the origin and development of ova. It should aid understanding of the crucial processes in organisms development.

Grauzone and Completion of Meiosis During Drosophila Oogenesis describes the work behind a major, award winning discovery: the establishment of a new pathway that specifically regulates the female meiosis, a process essential for sexual reproduction. This book chronicles a new gene mapping method and the cloning and documentation of several types of genes that were proven to have significant influence on the cell cycle. It is of interest to anyone doing work with fruit flies, both graduate students and principal investigators.

This volume covers the current knowledge base on the role of signaling and environmental pathways that control the normal development of germline stem cells, meiotic progression of oocytes, events of oocyte maturation and fertilization, and the birth of an embryo. Germ cells are uniquely poised to sustain life across generations through the fusion of oocyte and sperm. Because of the central importance of germ cells to life, much work has been dedicated to obtaining a clear understanding of the molecular and signaling events that control their formation and maintenance. Germ cells are set aside from somatic cells in the embryo and go through specialized meiotic cell cycles as the animal matures. These cell cycles are interspersed with long periods of arrest. In human females, meiosis I is initiated in the fetus. At birth, oocytes are arrested in meiosis I; after puberty, every month an oocyte initiates meiosis II – ovulation. Upon sperm availability these cells are fertilized, generate an embryo, and the cycle-of-life continues. During meiotic I progression and arrest, the fitness of oocytes and their progeny are likely influenced by environmental cues and signaling pathways. A lot of recent work has focused on understanding the mechanisms that regulate oocyte fitness and quality in humans and vertebrates. Much of our understanding on the events of meiosis I and germline stem cell populations comes from work in invertebrates, wherein the germline stem cells produce oocytes continuously through adult development. In both inverbrates and vertebrates nutritional and signaling pathways control the regulation of stem cells in such a manner so as to couple production of gametes with the nutritional availability. Additionally, mature oocytes arrest both in meiosis I and meiosis II, and signaling and nutritional pathways have been shown to regulate their formation, and maintenance, such that despite long periods of arrest, the oocyte quality is assured and errors in chromosome segregation and varied cytoplasmic events are minimal.

This volume provides current up-to-date protocols for preparing the ovary for various imaging techniques, genetic protocols for generating mutant clones, mosaic analysis and assessing cell death. Chapters address methods for performing genome wide gene expression analysis and bioinformatics for studies of RNA-protein interactions. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Drosophila Oogenesis: Methods and Protocols aims to ensure successful results in the further study of this vital field.