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In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes
Most researchers who study unicellular eukaryotes work with an extremely limited number of laboratory-adapted isolates that were obtained from the field decades ago, but the effects of passage in laboratory rodents, and adaptation to in vitro culture, have been little studied. For example, the vast...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8459984/ https://www.ncbi.nlm.nih.gov/pubmed/34516555 http://dx.doi.org/10.1371/journal.pntd.0009738 |
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author | Mulindwa, Julius Ssentamu, Geofrey Matovu, Enock Kamanyi Marucha, Kevin Aresta-Branco, Francisco Helbig, Claudia Clayton, Christine |
author_facet | Mulindwa, Julius Ssentamu, Geofrey Matovu, Enock Kamanyi Marucha, Kevin Aresta-Branco, Francisco Helbig, Claudia Clayton, Christine |
author_sort | Mulindwa, Julius |
collection | PubMed |
description | Most researchers who study unicellular eukaryotes work with an extremely limited number of laboratory-adapted isolates that were obtained from the field decades ago, but the effects of passage in laboratory rodents, and adaptation to in vitro culture, have been little studied. For example, the vast majority of studies of Trypanosoma brucei biology have concentrated on just two strains, Lister 427 and EATRO1125, which were taken from the field over half a century ago and have since have undergone innumerable passages in rodents and culture. We here describe two new Trypanosoma brucei brucei strains. MAK65 and MAK98, which have undergone only 3 rodent passages since isolation from Ugandan cattle. High-coverage sequencing revealed that adaptation of the parasites to culture was accompanied by changes in gene copy numbers. T. brucei has so far been considered to be uniformly diploid, but we also found trisomy of chromosome 5 not only in one Lister 427 culture, but also in the MAK98 field isolate. Trisomy of chromosome 6, and increased copies of other chromosome segments, were also seen in established cultured lines. The two new T. brucei strains should be useful to researchers interested in trypanosome differentiation and pathogenicity. Initial results suggested that the two strains have differing infection patterns in rodents. MAK65 is uniformly diploid and grew more reproducibly in bloodstream-form culture than MAK98. |
format | Online Article Text |
id | pubmed-8459984 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-84599842021-09-24 In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes Mulindwa, Julius Ssentamu, Geofrey Matovu, Enock Kamanyi Marucha, Kevin Aresta-Branco, Francisco Helbig, Claudia Clayton, Christine PLoS Negl Trop Dis Research Article Most researchers who study unicellular eukaryotes work with an extremely limited number of laboratory-adapted isolates that were obtained from the field decades ago, but the effects of passage in laboratory rodents, and adaptation to in vitro culture, have been little studied. For example, the vast majority of studies of Trypanosoma brucei biology have concentrated on just two strains, Lister 427 and EATRO1125, which were taken from the field over half a century ago and have since have undergone innumerable passages in rodents and culture. We here describe two new Trypanosoma brucei brucei strains. MAK65 and MAK98, which have undergone only 3 rodent passages since isolation from Ugandan cattle. High-coverage sequencing revealed that adaptation of the parasites to culture was accompanied by changes in gene copy numbers. T. brucei has so far been considered to be uniformly diploid, but we also found trisomy of chromosome 5 not only in one Lister 427 culture, but also in the MAK98 field isolate. Trisomy of chromosome 6, and increased copies of other chromosome segments, were also seen in established cultured lines. The two new T. brucei strains should be useful to researchers interested in trypanosome differentiation and pathogenicity. Initial results suggested that the two strains have differing infection patterns in rodents. MAK65 is uniformly diploid and grew more reproducibly in bloodstream-form culture than MAK98. Public Library of Science 2021-09-13 /pmc/articles/PMC8459984/ /pubmed/34516555 http://dx.doi.org/10.1371/journal.pntd.0009738 Text en © 2021 Mulindwa et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Mulindwa, Julius Ssentamu, Geofrey Matovu, Enock Kamanyi Marucha, Kevin Aresta-Branco, Francisco Helbig, Claudia Clayton, Christine In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
title | In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
title_full | In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
title_fullStr | In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
title_full_unstemmed | In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
title_short | In vitro culture of freshly isolated Trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
title_sort | in vitro culture of freshly isolated trypanosoma brucei brucei bloodstream forms results in gene copy-number changes |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8459984/ https://www.ncbi.nlm.nih.gov/pubmed/34516555 http://dx.doi.org/10.1371/journal.pntd.0009738 |
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