Characterization of Germ Line Development in Zebrafish PDF Download

Author: Arie Koen Braat
Publisher:
ISBN: 9789039320402
Category :
Languages : en
Pages : 121

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Author: Roland Dosch
Publisher:
ISBN: 9781071609705
Category : Cell differentiation
Languages : en
Pages : 383

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Book Description
This volume details a wide range of methods, ranging from beginner through advanced, used to further study zebrafish and fish germline. Chapter guide readers through cultivating and manipulating germ cells, imaging of germline processes and the molecular analysis of their, protein, and RNA. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Germline Development in the Zebrafish: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Author: Roland Dosch
Publisher: Humana
ISBN: 9781071609699
Category : Science
Languages : en
Pages : 383

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Book Description
This volume details a wide range of methods, ranging from beginner through advanced, used to further study zebrafish and fish germline.. Chapter guide readers through cultivating and manipulating germ cells, imaging of germline processes and the molecular analysis of their, protein, and RNA. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Germline Development in the Zebrafish: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Author: David Leu
Publisher:
ISBN: 9781124223117
Category :
Languages : en
Pages :

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Book Description
Genetic analysis of germ cell development in zebrafish has identified several germ cell autonomous genes that are required for different stages of germ cell development, such as nanos1, deadend, and ziwi, the homologue of the drosophila piwi. However, no germ cell non-autonomous factors that are required for germ cell maintenance have so far been described. Recently, the Draper lab has discovered that mutations in the gene encoding the Fibroblast growth factor (Fgf) ligand, Fgf24, results in the majority of animals being sterile. In addition to the sterility, all fgf24 mutant animals develop as phenotypic males, similar to that of ziwi mutants, suggesting that PGC or gonocyte survival may be impaired. The germline phenotype of fgf24 mutants suggests that fgf24 function is required in the gonad during larval development. The goal of the project is to determining if germ cells require an intact Fgf signaling pathway for their survival. To test the hypothesis that germ cells require Fgf signaling for their survival, I wanted to disrupt the Fgf signaling pathway by over expressing a dominant-negative Fgf receptor specifically in germ cells. However, no germ cell-specific promoter that drives gene expression in all germ cells including pre-meiotic germ cells has been described. It was therefore necessary to first identify such a promoter. In chapter 1, I cloned and tested the promoter activity of a 4.8 kb fragment upstream of ziwi, a gene known to be expressed in all germ cells, including pre-meiotic germ cells, I found that this fragment is capable to drive heterologous gene expression in all stages of germ cells. In chapter 2 I intended to determine if germ cells require an intact Fgf signaling pathway for their survival by overexpressing a dnFgfr in germ cells using the ziwi promoter. Unfortunately, because of mistakes made in construction of the dnfgfr transgenes, I have so far been unable to fully test the hypothesis.

Author: Christina Lauren Hansen
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

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In zebrafish (Danio rerio) and many other organisms, the differentiation of primordial germ cells (PGCs) requires the reliable transmission of maternally-derived germline determinants, collectively termed germ plasm. PGCs give rise to the germline; thus, the fidelity of PGC specification and development is intimately tied to the survival and fitness of sexually-reproducing species. In this dissertation, I systematically investigate the formation, structure, and composition of germ plasm aggregates in zebrafish and related fish species in the Danio and Devario genera. I provide a comprehensive overview of preformation (germ plasm-dependent) and induction (non-germ plasm) as mechanisms of PGC specification, including major findings in several model vertebrate species. PGC specification is also discussed in a phylogenetic context, with particular attention to conserved gene families and the ongoing efforts to characterize germline determination in non-model vertebrate species. In Chapters 3 and 4, I use advanced methods of fluorescence in situ hybridization (detailed in Appendix I), microscopy, and image analysis to characterize the formation and structure of zebrafish germ plasm aggregates at a macromolecular-level, from the identification of homotypic germline-determinant RNA particles (Chapter 3) to the description of and proposed mechanism for organized supramolecular assembly of germ plasm in cleavage-stage embryos (Chapter 4). Appendix II presents the creation and initial characterization of mutations in bucky ball 2-like, the only zebrafish paralog of the germ plasm organizing protein Bucky ball. In Chapter 5, I identify germ plasm in three non-zebrafish species within the Danionin clade: Danio kyathit, Danio albolineatus, and Devario aequipinnatus, and find that eleven RNAs, including several not previously reported as germ plasm components in any species beyond zebrafish, exhibit conserved localization patterns within the embryos of all three examined non-zebrafish species. One component of zebrafish germ plasm, ca15b, was observed in the germ plasm of the two non-zebrafish Danio species but lacked specific localization in Devario aequipinnatus embryos. Together, these works present an improved understanding of the molecular components and behavior of maternally-derived germ plasm in zebrafish and related species.

Author: Heide Schatten
Publisher: Springer Science & Business Media
ISBN: 1592597416
Category : Science
Languages : en
Pages : 421

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Book Description
The study of germ cells has undergone enormous advances in recent years and has entered into an explosive phase of new discoveries with the introd- tion of transgenic technologies and nuclear cloning. Basic knowledge and te- niques developed for lower vertebrate and invertebrate systems have facilitated the study of higher vertebrates, including humans. Many experiments that have first been performed on lower vertebrates provided the tools and strategies that could later be applied to other less readily available mammalian systems. The discovery of centrosomes in ascidians and sea urchin eggs now benefits st- ies of fertility and infertility in mammals including humans. External in vitro fertilization, now a common technique in assisted fertilization has only been possible as a result of numerous studies in lower systems in which external fertilization is natural. Egg activation, first explored in sea urchin and asc- ian eggs, now benefits cloning efficiency in farm and domestic animals. Gene manipulations and molecular methods have added to the possibilities of p- ducing live offspring with enormous biomedical, ecological, and economic implications. All sexually reproducing organisms produce primordial germ cells, a small population of cells that differentiate into gametes of either sex that carry to- potency, an ability to develop into an entire new organism. The two volumes on germ cells combine techniques in a variety of different systems and have selected those systems that have provided landmarks in advancing our kno- edge on germ cells.

Author: H. William Detrich III
Publisher: Academic Press
ISBN: 0123751616
Category : Science
Languages : en
Pages : 399

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Due to its prolific reproduction and the external development of the transparent embryo, the zebrafish is the prime model for genetic and developmental studies, as well as research in genomics. While genetically distant from humans, nonetheless the vertebrate zebrafish has comparable organs and tissues that make it the model organism for study of vertebrate development.This book, one of two new volumes in the Reliable Lab Solutions series dealing with zebrafish, brings together a robust and up-to-date collection of time-tested methods presented by the world’s leading scientists. Culled from previously published chapters in Methods in Cell Biology and updated by the original authors where relevant, it provides a comprehensive collection of protocols describing the most widely used techniques relevant to the study of zebrafish genetics and genomics. The methods in this volume were hand-selected by the editors, whose goal was to a provide a handy and cost-effective collection of fail-safe methods, tips, and "tricks of the trade" to both experienced researchers and more junior members in the lab. Provides busy researchers a quick reference for time-tested methods and protocols that really work, updated where possible by the original authors Gives pragmatic wisdom to the non-specialist from experts in the field with years of experience with trial and error

Author: Krasimir Ivanov Slanchev
Publisher:
ISBN:
Category :
Languages : en
Pages : 154

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Author: Uta Wolke
Publisher:
ISBN:
Category :
Languages : en
Pages : 144

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Author: Anne McLaren
Publisher: Cambridge University Press
ISBN: 9780521253291
Category : Science
Languages : en
Pages : 0

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