| Establishment of a high content assay for the identification and characterisation of bioactivities in crude bacterial extracts that interfere with the eukaryotic cell cycle. | |
| | |
MedLine Citation:
|
PMID: 19111838 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
High content microscopy as a screening tool to identify bioactive agents has provided researchers with the ability to characterise biological activities at the level of single cells. Here, we describe the development and the application of a high content screening assay for the identification and characterisation of cytostatic bioactivities from Myxobacteria extracts. In an automated microscopy assay Sf9 insect cells were visualised utilising the stains bisbenzimide Hoechst 33342, calcein AM, and propidium iodide. Imaging data were processed by the ScanR Analysis-software to determine the ploidy and vitality of each cell and to quantify cell populations. More than 98% of the Sf9 cells were viable and the culture consisted of diploid ( approximately 30%), tetraploid ( approximately 60%), polyploidic (<10%) and apoptotic (<5%) cells. Treatment with the reference substances blasticidin, colchicine, paclitaxel, and cytochalasin D induced changes in ploidy and vitality, which were characteristic for the respective bioactive substance. Furthermore, crude extracts from the chivosazole producing Myxobacterium Sorangium cellulosum So ce56 induced an increase of polyploid cells and a decrease in total cell count, while a mutant producing nearly no chivosazole triggered none of these effects. Purified chivosazole induced the same effects as the wild type extract. Similar effects have been observed for the reference compound cytochalasin D. On the basis of this assay, crude extracts of ten different Myxobacteria cultures were screened. Three extracts exhibited strong cytotoxic activities, further five extracts induced weak changes in the ploidy distribution, and two extracts showed no detectable effect within the assay. Therefore, this robust assay provides the ability to discover and characterise cytotoxic and cytostatic bioactivities in crude bacterial extracts. |
| | |
Authors:
|
Nickels A Jensen; Klaus Gerth; Tim Grotjohann; Dieter Kapp; Matthias Keck; Karsten Niehaus |
Related Documents
:
|
17994258 - Acrylamide synthesis using agar entrapped cells of rhodococcus rhodochrous pa-34 in a p... 10653768 - Survival of bifidobacterium longum immobilized in calcium alginate beads in simulated g... 2295608 - Pyridoxine-derived b6 vitamers and pyridoxal 5'-phosphate-binding proteins in cytosolic... 7217008 - Metabolism of resorcinylic compounds by bacteria: new pathway for resorcinol catabolism... 1254928 - Studies of cellular differentiation by automated cell separation. two model systems: fr... 1463598 - Functional and biochemical characterization of osteoclast-like cells derived from giant... |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-12-09 |
Journal Detail:
|
Title: Journal of biotechnology Volume: 140 ISSN: 1873-4863 ISO Abbreviation: J. Biotechnol. Publication Date: 2009 Mar |
Date Detail:
|
Created Date: 2009-03-16 Completed Date: 2009-08-24 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 8411927 Medline TA: J Biotechnol Country: Netherlands |
Other Details:
|
Languages: eng Pagination: 124-34 Citation Subset: IM |
Affiliation:
|
Department of Proteome and Metabolome Research, Faculty of Biology, Bielefeld University, Bielefeld, Germany. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Cell Cycle / drug effects Cell Line Cell Proliferation / drug effects Colchicine / pharmacology Coloring Agents / metabolism Complex Mixtures / pharmacology* Cytochalasin D / pharmacology Cytostatic Agents / pharmacology* Drug Discovery Fluorescent Dyes / metabolism Image Processing, Computer-Assisted / methods* Macrolides / pharmacology Microscopy / methods* Myxococcales / metabolism* Paclitaxel / pharmacology Ploidies Pyrrolidinones / pharmacology |
| Chemical | |
Reg. No./Substance:
|
0/Coloring Agents; 0/Complex Mixtures; 0/Cytostatic Agents; 0/Fluorescent Dyes; 0/Macrolides; 0/Pyrrolidinones; 0/blasticidin A; 0/chivosazol A; 22144-77-0/Cytochalasin D; 33069-62-4/Paclitaxel; 64-86-8/Colchicine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Activity of the AtMRP3 promoter in transgenic Arabidopsis thaliana and Nicotiana tabacum plants is i...
Next Document: Pathological, immunohistochemical and bacteriological study of tissues and milk of cows and fetuses ...