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The Effect of In Vitro Cultivation on the Transcriptome of Adult Brugia malayi

Identifieur interne : 007D08 ( Ncbi/Merge ); précédent : 007D07; suivant : 007D09

The Effect of In Vitro Cultivation on the Transcriptome of Adult Brugia malayi

Auteurs : Cristina Ballesteros [Canada] ; Lucienne Tritten [Canada] ; Maeghan O Eill [Canada] ; Erica Burkman [États-Unis] ; Weam I. Zaky [États-Unis] ; Jianguo Xia [Canada] ; Andrew Moorhead [États-Unis] ; Steven A. Williams [États-Unis] ; Timothy G. Geary [Canada]

Source :

RBID : PMC:4699822

Abstract

Background

Filarial nematodes cause serious and debilitating infections in human populations of tropical countries, contributing to an entrenched cycle of poverty. Only one human filarial parasite, Brugia malayi, can be maintained in rodents in the laboratory setting. It has been a widely used model organism in experiments that employ culture systems, the impact of which on the worms is unknown.

Methodology/Principal Findings

Using Illumina RNA sequencing, we characterized changes in gene expression upon in vitro maintenance of adult B. malayi female worms at four time points: immediately upon removal from the host, immediately after receipt following shipment, and after 48 h and 5 days in liquid culture media. The dramatic environmental change and the 24 h time lapse between removal from the host and establishment in culture caused a globally dysregulated gene expression profile. We found a maximum of 562 differentially expressed genes based on pairwise comparison between time points. After an initial shock upon removal from the host and shipping, a few stress fingerprints remained after 48 h in culture and until the experiment was stopped. This was best illustrated by a strong and persistent up-regulation of several genes encoding cuticle collagens, as well as serpins.

Conclusions/Significance

These findings suggest that B. malayi can be maintained in culture as a valid system for pharmacological and biological studies, at least for several days after removal from the host and adaptation to the new environment. However, genes encoding several stress indicators remained dysregulated until the experiment was stopped.


Url:
DOI: 10.1371/journal.pntd.0004311
PubMed: 26727204
PubMed Central: 4699822

Links toward previous steps (curation, corpus...)


Links to Exploration step

PMC:4699822

Le document en format XML

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<p>Filarial nematodes cause serious and debilitating infections in human populations of tropical countries, contributing to an entrenched cycle of poverty. Only one human filarial parasite,
<italic>Brugia malayi</italic>
, can be maintained in rodents in the laboratory setting. It has been a widely used model organism in experiments that employ culture systems, the impact of which on the worms is unknown.</p>
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<p>Using Illumina RNA sequencing, we characterized changes in gene expression upon
<italic>in vitro</italic>
maintenance of adult
<italic>B</italic>
.
<italic>malayi</italic>
female worms at four time points: immediately upon removal from the host, immediately after receipt following shipment, and after 48 h and 5 days in liquid culture media. The dramatic environmental change and the 24 h time lapse between removal from the host and establishment in culture caused a globally dysregulated gene expression profile. We found a maximum of 562 differentially expressed genes based on pairwise comparison between time points. After an initial shock upon removal from the host and shipping, a few stress fingerprints remained after 48 h in culture and until the experiment was stopped. This was best illustrated by a strong and persistent up-regulation of several genes encoding cuticle collagens, as well as serpins.</p>
</sec>
<sec id="sec003">
<title>Conclusions/Significance</title>
<p>These findings suggest that
<italic>B</italic>
.
<italic>malayi</italic>
can be maintained in culture as a valid system for pharmacological and biological studies, at least for several days after removal from the host and adaptation to the new environment. However, genes encoding several stress indicators remained dysregulated until the experiment was stopped.</p>
</sec>
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</TEI>
<pmc article-type="research-article">
<pmc-dir>properties open_access</pmc-dir>
<front>
<journal-meta>
<journal-id journal-id-type="nlm-ta">PLoS Negl Trop Dis</journal-id>
<journal-id journal-id-type="iso-abbrev">PLoS Negl Trop Dis</journal-id>
<journal-id journal-id-type="publisher-id">plos</journal-id>
<journal-id journal-id-type="pmc">plosntds</journal-id>
<journal-title-group>
<journal-title>PLoS Neglected Tropical Diseases</journal-title>
</journal-title-group>
<issn pub-type="ppub">1935-2727</issn>
<issn pub-type="epub">1935-2735</issn>
<publisher>
<publisher-name>Public Library of Science</publisher-name>
<publisher-loc>San Francisco, CA USA</publisher-loc>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="pmid">26727204</article-id>
<article-id pub-id-type="pmc">4699822</article-id>
<article-id pub-id-type="doi">10.1371/journal.pntd.0004311</article-id>
<article-id pub-id-type="publisher-id">PNTD-D-15-01544</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Research Article</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>The Effect of
<italic>In Vitro</italic>
Cultivation on the Transcriptome of Adult
<italic>Brugia malayi</italic>
</article-title>
<alt-title alt-title-type="running-head">Transcriptome of
<italic>In Vitro</italic>
Cultured
<italic>Brugia malayi</italic>
</alt-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" equal-contrib="yes">
<name>
<surname>Ballesteros</surname>
<given-names>Cristina</given-names>
</name>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author" equal-contrib="yes">
<name>
<surname>Tritten</surname>
<given-names>Lucienne</given-names>
</name>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>O’Neill</surname>
<given-names>Maeghan</given-names>
</name>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Burkman</surname>
<given-names>Erica</given-names>
</name>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Zaky</surname>
<given-names>Weam I.</given-names>
</name>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
<xref ref-type="aff" rid="aff004">
<sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Xia</surname>
<given-names>Jianguo</given-names>
</name>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Moorhead</surname>
<given-names>Andrew</given-names>
</name>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Williams</surname>
<given-names>Steven A.</given-names>
</name>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
<xref ref-type="aff" rid="aff004">
<sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Geary</surname>
<given-names>Timothy G.</given-names>
</name>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref rid="cor001" ref-type="corresp">*</xref>
</contrib>
</contrib-group>
<aff id="aff001">
<label>1</label>
<addr-line>Institute of Parasitology, Centre for Host-Parasite Interactions, McGill University, Sainte-Anne-de-Bellevue, Quebec, Canada</addr-line>
</aff>
<aff id="aff002">
<label>2</label>
<addr-line>Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America</addr-line>
</aff>
<aff id="aff003">
<label>3</label>
<addr-line>Filariasis Research Reagent Resource Center, Northampton, Massachusetts, United States of America</addr-line>
</aff>
<aff id="aff004">
<label>4</label>
<addr-line>Department of Biological Sciences, Smith College, Northampton, Massachusetts, United States of America</addr-line>
</aff>
<contrib-group>
<contrib contrib-type="editor">
<name>
<surname>Ghedin</surname>
<given-names>Elodie</given-names>
</name>
<role>Editor</role>
<xref ref-type="aff" rid="edit1"></xref>
</contrib>
</contrib-group>
<aff id="edit1">
<addr-line>New York University, UNITED STATES</addr-line>
</aff>
<author-notes>
<fn fn-type="conflict" id="coi001">
<p>The authors have declared that no competing interests exist.</p>
</fn>
<fn fn-type="con" id="contrib001">
<p>Conceived and designed the experiments: CB LT MO TGG. Performed the experiments: CB LT MO EB WIZ. Analyzed the data: CB LT MO EB JX. Contributed reagents/materials/analysis tools: EB SAW AM. Wrote the paper: CB LT JX TGG AM.</p>
</fn>
<corresp id="cor001">* E-mail:
<email>timothy.g.geary@mcgill.ca</email>
</corresp>
</author-notes>
<pub-date pub-type="epub">
<day>4</day>
<month>1</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="collection">
<month>1</month>
<year>2016</year>
</pub-date>
<volume>10</volume>
<issue>1</issue>
<elocation-id>e0004311</elocation-id>
<history>
<date date-type="received">
<day>1</day>
<month>9</month>
<year>2015</year>
</date>
<date date-type="accepted">
<day>29</day>
<month>11</month>
<year>2015</year>
</date>
</history>
<permissions>
<copyright-statement>© 2016 Ballesteros et al</copyright-statement>
<copyright-year>2016</copyright-year>
<copyright-holder>Ballesteros et al</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<license-p>This is an open access article distributed under the terms of the
<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>
, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited</license-p>
</license>
</permissions>
<self-uri content-type="pdf" xlink:type="simple" xlink:href="pntd.0004311.pdf"></self-uri>
<abstract>
<sec id="sec001">
<title>Background</title>
<p>Filarial nematodes cause serious and debilitating infections in human populations of tropical countries, contributing to an entrenched cycle of poverty. Only one human filarial parasite,
<italic>Brugia malayi</italic>
, can be maintained in rodents in the laboratory setting. It has been a widely used model organism in experiments that employ culture systems, the impact of which on the worms is unknown.</p>
</sec>
<sec id="sec002">
<title>Methodology/Principal Findings</title>
<p>Using Illumina RNA sequencing, we characterized changes in gene expression upon
<italic>in vitro</italic>
maintenance of adult
<italic>B</italic>
.
<italic>malayi</italic>
female worms at four time points: immediately upon removal from the host, immediately after receipt following shipment, and after 48 h and 5 days in liquid culture media. The dramatic environmental change and the 24 h time lapse between removal from the host and establishment in culture caused a globally dysregulated gene expression profile. We found a maximum of 562 differentially expressed genes based on pairwise comparison between time points. After an initial shock upon removal from the host and shipping, a few stress fingerprints remained after 48 h in culture and until the experiment was stopped. This was best illustrated by a strong and persistent up-regulation of several genes encoding cuticle collagens, as well as serpins.</p>
</sec>
<sec id="sec003">
<title>Conclusions/Significance</title>
<p>These findings suggest that
<italic>B</italic>
.
<italic>malayi</italic>
can be maintained in culture as a valid system for pharmacological and biological studies, at least for several days after removal from the host and adaptation to the new environment. However, genes encoding several stress indicators remained dysregulated until the experiment was stopped.</p>
</sec>
</abstract>
<abstract abstract-type="summary">
<title>Author Summary</title>
<p>Infections with filarial worms cause serious physical impairment and affect tens of millions of people in tropical and subtropical countries. To better understand the biology and pharmacology of these parasites,
<italic>Brugia malayi</italic>
is often used as a model. This parasite can be maintained in the laboratory in Mongolian jirds, enabling researchers to test drugs
<italic>in vivo</italic>
and
<italic>in vitro</italic>
, among other studies. The effects of removing worms from their hosts and culturing them may affect many aspects of their physiology, including response to drugs, but the extent to which the worms undergo changes during culture has remained unknown. Using deep RNA sequencing and bioinformatics tools, we examined the global transcriptomic profile of
<italic>B</italic>
.
<italic>malayi</italic>
females at four different time points over 5 days in culture. Focusing on genes that are differentially expressed at various time points, we observed a general perturbation of the expression profile between dissection from the host and receipt after shipment. The expression of several genes remained changed at the end of the experiment, after 5 days under controlled conditions; in particular, genes encoding cuticle collagens were prominently represented and strongly overexpressed.</p>
</abstract>
<funding-group>
<funding-statement>LT received a Swiss National Science Foundation Postdoctoral Fellowship and a Canadian Institutes of Health Research Postdoctoral Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement>
</funding-group>
<counts>
<fig-count count="5"></fig-count>
<table-count count="3"></table-count>
<page-count count="20"></page-count>
</counts>
<custom-meta-group>
<custom-meta id="data-availability">
<meta-name>Data Availability</meta-name>
<meta-value>All relevant data are within the paper and its Supporting Information files, except for the data deposited in the GEO database at the following link:
<ext-link ext-link-type="uri" xlink:href="http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=exuheqgyrhepnev&acc=GSE72592">http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=exuheqgyrhepnev&acc=GSE72592</ext-link>
</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
<notes>
<title>Data Availability</title>
<p>All relevant data are within the paper and its Supporting Information files, except for the data deposited in the GEO database at the following link:
<ext-link ext-link-type="uri" xlink:href="http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=exuheqgyrhepnev&acc=GSE72592">http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=exuheqgyrhepnev&acc=GSE72592</ext-link>
</p>
</notes>
</front>
</pmc>
<affiliations>
<list>
<country>
<li>Canada</li>
<li>États-Unis</li>
</country>
<region>
<li>Géorgie (États-Unis)</li>
<li>Massachusetts</li>
<li>Québec</li>
</region>
<settlement>
<li>Montréal</li>
</settlement>
<orgName>
<li>Université McGill</li>
</orgName>
</list>
<tree>
<country name="Canada">
<region name="Québec">
<name sortKey="Ballesteros, Cristina" sort="Ballesteros, Cristina" uniqKey="Ballesteros C" first="Cristina" last="Ballesteros">Cristina Ballesteros</name>
</region>
<name sortKey="Geary, Timothy G" sort="Geary, Timothy G" uniqKey="Geary T" first="Timothy G." last="Geary">Timothy G. Geary</name>
<name sortKey="O Eill, Maeghan" sort="O Eill, Maeghan" uniqKey="O Eill M" first="Maeghan" last="O Eill">Maeghan O Eill</name>
<name sortKey="Tritten, Lucienne" sort="Tritten, Lucienne" uniqKey="Tritten L" first="Lucienne" last="Tritten">Lucienne Tritten</name>
<name sortKey="Xia, Jianguo" sort="Xia, Jianguo" uniqKey="Xia J" first="Jianguo" last="Xia">Jianguo Xia</name>
</country>
<country name="États-Unis">
<region name="Géorgie (États-Unis)">
<name sortKey="Burkman, Erica" sort="Burkman, Erica" uniqKey="Burkman E" first="Erica" last="Burkman">Erica Burkman</name>
</region>
<name sortKey="Burkman, Erica" sort="Burkman, Erica" uniqKey="Burkman E" first="Erica" last="Burkman">Erica Burkman</name>
<name sortKey="Moorhead, Andrew" sort="Moorhead, Andrew" uniqKey="Moorhead A" first="Andrew" last="Moorhead">Andrew Moorhead</name>
<name sortKey="Moorhead, Andrew" sort="Moorhead, Andrew" uniqKey="Moorhead A" first="Andrew" last="Moorhead">Andrew Moorhead</name>
<name sortKey="Williams, Steven A" sort="Williams, Steven A" uniqKey="Williams S" first="Steven A." last="Williams">Steven A. Williams</name>
<name sortKey="Williams, Steven A" sort="Williams, Steven A" uniqKey="Williams S" first="Steven A." last="Williams">Steven A. Williams</name>
<name sortKey="Zaky, Weam I" sort="Zaky, Weam I" uniqKey="Zaky W" first="Weam I." last="Zaky">Weam I. Zaky</name>
<name sortKey="Zaky, Weam I" sort="Zaky, Weam I" uniqKey="Zaky W" first="Weam I." last="Zaky">Weam I. Zaky</name>
</country>
</tree>
</affiliations>
</record>

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