Ludwig Eichinger Francisco Rivero

Slug Phototaxis, Thermotaxis, and Spontaneous Turning Behavior Paul R. Fisher and Sarah J. Annesley Summary Dictyostelium slug phototaxis and thermotaxis are readily assayed phenotypes that reflect with great sensitivity and specificity the interactions of environmental stimuli with morphogenetic signaling systems controlling the collective movement of slug cells. Methods are described for conducting and recording phototaxis, thermotaxis, and spontaneous turning experiments, although it is...

Info

Result for searching ddv41j23* using the Genome Browser's Landmark or Region box. The window shows that two clones were found, ddv41j23.3 and ddv41j23.5, in adjacent locations on chromosome 5. Clicking on the diamond next to the clone name or on the coordinates leads to exactly that region on the chromosome in the Genome Browser window. In a similar fashion, this tool can be used to search genes with similar name symbols such as gpa* or cpn*. http dictyBase.org db The Genome Browser...

Humana Press Totowa New Jersey

2006 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. Methods in Molecular Biology is a trademark of The Humana Press Inc. All papers, comments, opinions, conclusions, or recommendations are those of the author(s),...

Investigating Gene Expression

In Situ Hybridization and Reporter Genes Ricardo Escalante and Leandro Sastre Summary Coordinated cell type differentiation is essential for morphogenesis during Dictyostelium development. The specification of different cell types and the regulation of temporal and spatial patterns of expression of cell type-specific genes are important problems currently being addressed in many laboratories. Besides, determination of gene expression patterns provides significant information in the...

Under Agarose Chemotaxis of Dictyostelium discoideum

In the vegetative state, Dictyostelium amoebae are chemotactic toward pterins released by bacteria, whereas during multicellular development, they become chemotac-tic to endogenously produced cAMP. A variety of assays have been used to visualize and quantify chemotactic movement. Under-agarose chemotaxis provides a simple and flexible assay that permits high-resolution imaging and quantification of the motility behavior of individual cells and populations by both transmitted light and...

The Secret Lives of Dictyostelium

Dictyostelium Discoideum

Research on Dictyostelium has a long history, the trend of which has been to add cell biology, natural products, and evolutionary biology research to the traditional studies of development. The methods presented here and the recent publication of the genome present vast new opportunities. Key Words Cellular slime mold natural products soil ameba amoeboid genome. In 1991, the archives of the Soviet Union and its satellites began to open and to inform us in the West about events that we had...

Optimized Fixation and Immunofluorescence Staining Methods for Dictyostelium Cells

Draw The Setup For Fixation

Neuhaus, and Thierry Soldati Summary Recent years have seen a powerful revival of fluorescence microscopy techniques, both to observe live cells and fixed objects. The limits of sensitivity, simultaneous detection of multiple chromophores, and spatial resolution have all been pushed to the extreme. Therefore, it is essential to improve in parallel the quality of the structural and antigenic preservation during fixation and immunostaining. Chemical fixations are broadly...

Application of 2D and 3D DIAS to Motion Analysis of Live Cells in Transmission and Confocal Microscopy Imaging

Deborah Wessels, Spencer Kuhl, and David R. Soll Summary The chemotactic signal in Dictyostelium is a cAMP wave that is relayed over relatively large distances through a cell population during aggregation. Cells exhibit unique behaviors in response to the different spatial, temporal, and concentration components of the cAMP wave, suggesting that distinct signal transduction pathways are evoked in each of the various phases of the wave. For this reason, we designed a set of experimental...

Generation of Multiple Knockout Mutants Using the CreloxP System

Streak Plate Gene Knocouts

Dictyostelium discoideum is an exceptionally powerful system by which to study various aspects of modern cell and developmental biology. The completion of the genome sequencing project together with a high-efficiency of targeted gene disruption have enabled researchers to characterize many specific gene functions. However, as a result of many gene products with overlapping functions, there is great need to produce mutants carrying mutations in multiple genes. We have, therefore, developed a...

Restriction Enzyme Mediated Integration REMI Mutagenesis

A method for the integration of linear DNA into the Dictyostelium genome is described. Restriction enzyme-mediated integration, or REMI, involves the transformation of cells with a mixture of plasmid DNA, linearized with a restriction enzyme, along with a restriction enzyme that is capable of generating compatible cohesive ends in the genome. The enzyme stimulates integration of the DNA into cognate restriction sites in the chromosomes, usually as a single-copy insertion event and with little...

Proteomic Analysis of Dictyostelium discoideum

Udo Roth, Stefan M ller, and Franz-Georg Hanisch The social amoeba Dictyostelium discoideum is already known as a model organism for a variety of cellular and molecular studies. Now that the genome sequencing project has been completed and different tools with which to overexpress or knock out genes are available, this species has also moved into the spotlight of functional genomics studies. Consequently, this genomic sequence information can now be exploited to realize D. discoideum proteomics...