Cargando…
The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation
BACKGROUND: Flatworms are characterized by an outstanding stem cell system. These stem cells (neoblasts) can give rise to all cell types including germ cells and power the exceptional regenerative capacity of many flatworm species. Macrostomum lignano is an emerging model system to study stem cell b...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2007
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1828727/ https://www.ncbi.nlm.nih.gov/pubmed/17349046 http://dx.doi.org/10.1186/1742-9994-4-9 |
_version_ | 1782132741039456256 |
---|---|
author | Pfister, Daniela De Mulder, Katrien Philipp, Isabelle Kuales, Georg Hrouda, Martina Eichberger, Paul Borgonie, Gaetan Hartenstein, Volker Ladurner, Peter |
author_facet | Pfister, Daniela De Mulder, Katrien Philipp, Isabelle Kuales, Georg Hrouda, Martina Eichberger, Paul Borgonie, Gaetan Hartenstein, Volker Ladurner, Peter |
author_sort | Pfister, Daniela |
collection | PubMed |
description | BACKGROUND: Flatworms are characterized by an outstanding stem cell system. These stem cells (neoblasts) can give rise to all cell types including germ cells and power the exceptional regenerative capacity of many flatworm species. Macrostomum lignano is an emerging model system to study stem cell biology of flatworms. It is complementary to the well-studied planarians because of its small size, transparency, simple culture maintenance, the basal taxonomic position and its less derived embryogenesis that is more closely related to spiralians. The development of cell-, tissue- and organ specific markers is necessary to further characterize the differentiation potential of flatworm stem cells. Large scale in situ hybridization is a suitable tool to identify possible markers. Distinguished genes identified in a large scale screen in combination with manipulation of neoblasts by hydroxyurea or irradiation will advance our understanding of differentiation and regulation of the flatworm stem cell system. RESULTS: We have set up a protocol for high throughput large scale whole mount in situ hybridization for the flatworm Macrostomum lignano. In the pilot screen, a number of cell-, tissue- or organ specific expression patterns were identified. We have selected two stem cell- and germ cell related genes – macvasa and macpiwi – and studied effects of hydroxyurea (HU) treatment or irradiation on gene expression. In addition, we have followed cell proliferation using a mitosis marker and bromodeoxyuridine labeling of S-phase cells after various periods of HU exposure or different irradiation levels. HU mediated depletion of cell proliferation and HU induced reduction of gene expression was used to generate a cDNA library by suppressive subtractive hybridization. 147 differentially expressed genes were sequenced and assigned to different categories. CONCLUSION: We show that Macrostomum lignano is a suitable organism to perform high throughput large scale whole mount in situ hybridization. Genes identified in such screens – together with BrdU/H3 labeling – can be used to obtain information on flatworm neoblasts. |
format | Text |
id | pubmed-1828727 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-18287272007-03-20 The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation Pfister, Daniela De Mulder, Katrien Philipp, Isabelle Kuales, Georg Hrouda, Martina Eichberger, Paul Borgonie, Gaetan Hartenstein, Volker Ladurner, Peter Front Zool Research BACKGROUND: Flatworms are characterized by an outstanding stem cell system. These stem cells (neoblasts) can give rise to all cell types including germ cells and power the exceptional regenerative capacity of many flatworm species. Macrostomum lignano is an emerging model system to study stem cell biology of flatworms. It is complementary to the well-studied planarians because of its small size, transparency, simple culture maintenance, the basal taxonomic position and its less derived embryogenesis that is more closely related to spiralians. The development of cell-, tissue- and organ specific markers is necessary to further characterize the differentiation potential of flatworm stem cells. Large scale in situ hybridization is a suitable tool to identify possible markers. Distinguished genes identified in a large scale screen in combination with manipulation of neoblasts by hydroxyurea or irradiation will advance our understanding of differentiation and regulation of the flatworm stem cell system. RESULTS: We have set up a protocol for high throughput large scale whole mount in situ hybridization for the flatworm Macrostomum lignano. In the pilot screen, a number of cell-, tissue- or organ specific expression patterns were identified. We have selected two stem cell- and germ cell related genes – macvasa and macpiwi – and studied effects of hydroxyurea (HU) treatment or irradiation on gene expression. In addition, we have followed cell proliferation using a mitosis marker and bromodeoxyuridine labeling of S-phase cells after various periods of HU exposure or different irradiation levels. HU mediated depletion of cell proliferation and HU induced reduction of gene expression was used to generate a cDNA library by suppressive subtractive hybridization. 147 differentially expressed genes were sequenced and assigned to different categories. CONCLUSION: We show that Macrostomum lignano is a suitable organism to perform high throughput large scale whole mount in situ hybridization. Genes identified in such screens – together with BrdU/H3 labeling – can be used to obtain information on flatworm neoblasts. BioMed Central 2007-03-09 /pmc/articles/PMC1828727/ /pubmed/17349046 http://dx.doi.org/10.1186/1742-9994-4-9 Text en Copyright © 2007 Pfister et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Pfister, Daniela De Mulder, Katrien Philipp, Isabelle Kuales, Georg Hrouda, Martina Eichberger, Paul Borgonie, Gaetan Hartenstein, Volker Ladurner, Peter The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
title | The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
title_full | The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
title_fullStr | The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
title_full_unstemmed | The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
title_short | The exceptional stem cell system of Macrostomum lignano: Screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
title_sort | exceptional stem cell system of macrostomum lignano: screening for gene expression and studying cell proliferation by hydroxyurea treatment and irradiation |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1828727/ https://www.ncbi.nlm.nih.gov/pubmed/17349046 http://dx.doi.org/10.1186/1742-9994-4-9 |
work_keys_str_mv | AT pfisterdaniela theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT demulderkatrien theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT philippisabelle theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT kualesgeorg theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT hroudamartina theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT eichbergerpaul theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT borgoniegaetan theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT hartensteinvolker theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT ladurnerpeter theexceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT pfisterdaniela exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT demulderkatrien exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT philippisabelle exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT kualesgeorg exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT hroudamartina exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT eichbergerpaul exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT borgoniegaetan exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT hartensteinvolker exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation AT ladurnerpeter exceptionalstemcellsystemofmacrostomumlignanoscreeningforgeneexpressionandstudyingcellproliferationbyhydroxyureatreatmentandirradiation |