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N-arachidonoyl glycine, an abundant endogenous lipid, potently drives directed cellular migration through GPR18, the putative abnormal cannabidiol receptor - PubMed

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The data generated from this study supports the hypothesis that GPR18 is the previously unidentified &#x27;Abn-CBD&#x27; receptor. 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data-collections-open-panel-url-hash="#open-collections-panel"> <div class="inner-wrap"> <h3 class="action-panel-heading"> Add to Collections </h3> <form id="collections-action-panel-form" class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors" data-existing-collections-url="/list-existing-collections/" data-add-to-existing-collection-url="/add-to-existing-collection/" data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/" data-get-article-ids-by-search-url="/get-article-ids-by-search/" data-myncbi-max-collection-name-length="100" data-add-to-collection-max-amount="1000" data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"> <input type="hidden" name="csrfmiddlewaretoken" value="6UE9QyRSrkKuEXMZCVs9T4Yd9SaEeMVWhCCQ4JIA6c5IN667p5FBKHC2S7nAAMBj"> <div class="choice-group" role="radiogroup"> <ul class="radio-group-items"> <li> <input type="radio" id="collections-action-panel-new" class="collections-new" name="collections" value="new" data-ga-category="save_share" data-ga-action="collections" data-ga-label="collections_radio_new"> <label for="collections-action-panel-new">Create a new collection</label> </li> <li> <input type="radio" id="collections-action-panel-existing" class="collections-existing" name="collections" value="existing" checked="true" data-ga-category="save_share" data-ga-action="collections" data-ga-label="collections_radio_existing"> <label for="collections-action-panel-existing">Add to an existing collection</label> </li> </ul> </div> <div class="controls-wrapper"> <div class="action-panel-control-wrap new-collections-controls"> <label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk"> Name your collection: </label> <input type="text" name="add-to-new-collection" id="collections-action-panel-add-to-new" class="collections-action-add-to-new" pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/" maxlength="100" data-ga-category="save_share" data-ga-action="create_collection" data-ga-label="non_favorties_collection"> <div class="collections-new-name-too-long usa-input-error-message selection-validation-message"> Name must be less than 100 characters </div> </div> <div class="action-panel-control-wrap existing-collections-controls"> <label for="collections-action-panel-add-to-existing" class="action-panel-label"> Choose a collection: </label> <select id="collections-action-panel-add-to-existing" class="action-panel-selector collections-action-add-to-existing" data-ga-category="save_share" data-ga-action="select_collection" data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'"> </select> <div class="collections-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your collection due to an error<br> <a href="#">Please try again</a> </div> </div> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore data-ga-category="save_share" data-ga-action="collections" data-ga-label="add"> Add </button> <button class="action-panel-cancel" aria-label="Close 'Add to Collections' panel" ref="linksrc=close_collections_panel" aria-controls="collections-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="collections" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="bibliography-action-panel" class="bibliography-action-panel action-panel in-progress-dots-panel" aria-hidden="true" data-bibliography-open-panel-enabled="false" data-bibliography-open-panel-url-hash="#open-bibliography-panel"> <div class="inner-wrap"> <h3 class="action-panel-heading"> Add to My Bibliography </h3> <form id="bibliography-action-panel-form" class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors" data-add-to-bibliography-max-amount="100" data-add-to-bibliography-batch-size="10" data-bibliography-delegates-url="/list-bibliography-delegates/" data-add-to-bibliography-url="/add-to-bibliography/" data-get-article-ids-by-search-url="/get-article-ids-by-search/" data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/"> <input type="hidden" name="csrfmiddlewaretoken" value="6UE9QyRSrkKuEXMZCVs9T4Yd9SaEeMVWhCCQ4JIA6c5IN667p5FBKHC2S7nAAMBj"> <div class="action-panel-control-wrap bibliographies-controls"> <div class="choice-group"> <ul class="bibliographies-action-add radio-group-items"> <li> <input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/> <label for="my-bibliography">My Bibliography</label> </li> </ul> </div> </div> <div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your delegates due to an error<br> <a href="#">Please try again</a> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore> Add </button> <button class="action-panel-cancel" aria-label="Close 'Add to bibliography' panel" ref="linksrc=close_bibliography_panel" aria-controls="bibliography-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="mybib" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true" data-saved-search-open-panel-enabled="false" data-saved-search-open-panel-url-hash="#open-saved-search-panel"> <div class="inner-wrap"> <h2 class="action-panel-heading"> Your saved search </h2> <form id="saved-search-action-panel-form" class="saved-search-action-panel-form action-panel-content action-form" data-create-saved-search-url="/create-saved-search/" data-try-search-terms-url="/try-search-term/" data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/"> <input type="hidden" name="csrfmiddlewaretoken" value="6UE9QyRSrkKuEXMZCVs9T4Yd9SaEeMVWhCCQ4JIA6c5IN667p5FBKHC2S7nAAMBj"> <div class="action-panel-control-wrap"> <label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk"> Name of saved search: </label> <input maxlength="200" type="text" name="saved-search-name" id="saved-search-name" class="saved-search-name" value="" required pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"> </div> <div class="action-panel-control-wrap"> <label for="saved-search-term" class="action-panel-label required-field-asterisk"> Search terms: </label> <textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea> </div> <div class="test-search-term-wrap"> <a href="#" class="try-search-term">Test search terms</a> </div> <div class="choice-group action-panel-extra-margin-top"> <span class="action-panel-label" id="fieldset-label"> Would you like email updates of new search results? </span> <fieldset id="saved-search-alert" aria-describedby="fieldset-label"> <legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend> <ul class="radio-group-items"> <li> <input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked> <label for="saved-search-alert-yes" class="action-panel-label">Yes</label> </li> <li> <input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no"> <label for="saved-search-alert-no" class="action-panel-label">No</label> </li> </ul> </fieldset> </div> <div class="alert-schedule-wrap"> <div class="action-panel-control-wrap"> <label class="action-panel-label"> Email: </label> <span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span> </div> <div class="action-panel-control-wrap action-panel-extra-margin-top"> <label for="saved-search-frequency" class="action-panel-label"> Frequency: </label> <select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency"> <option value="monthly">Monthly</option> <option value="weekly">Weekly</option> <option value="daily">Daily</option> </select> </div> <div class="action-panel-control-wrap saved-search-monthly-additional"> <label for="saved-search-monthly-on-day" class="action-panel-label"> Which day? </label> <select id="saved-search-monthly-on-day" class="no-border-panel-selector"> <option value="Sunday">The first Sunday</option> <option value="Monday">The first Monday</option> <option value="Tuesday">The first Tuesday</option> <option value="Wednesday">The first Wednesday</option> <option value="Thursday">The first Thursday</option> <option value="Friday">The first Friday</option> <option value="Saturday">The first Saturday</option> <option value="day">The first day</option> <option value="weekday">The first weekday</option> </select> </div> <div class="action-panel-control-wrap saved-search-weekly-additional"> <label for="saved-search-weekly-on-day" class="action-panel-label"> Which day? </label> <select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day"> <option value="Sunday">Sunday</option> <option value="Monday">Monday</option> <option value="Tuesday">Tuesday</option> <option value="Wednesday">Wednesday</option> <option value="Thursday">Thursday</option> <option value="Friday">Friday</option> <option value="Saturday">Saturday</option> </select> </div> <div class="action-panel-control-wrap"> <label for="saved-search-report" class="action-panel-label"> Report format: </label> <select id="saved-search-report" class="no-border-panel-selector saved-search-report"> <option value="DocSum">Summary</option> <option value="DocSumText">Summary (text)</option> <option value="Abstract">Abstract</option> <option value="AbstractText">Abstract (text)</option> <option value="MEDLINE">PubMed</option> </select> </div> <div class="action-panel-control-wrap"> <label for="saved-search-amount" class="action-panel-label"> Send at most: </label> <select id="saved-search-amount" class="no-border-panel-selector saved-search-amount"> <option value="1">1 item</option> <option value="5" selected>5 items</option> <option value="10">10 items</option> <option value="20">20 items</option> <option value="50">50 items</option> <option value="100">100 items</option> <option value="200">200 items</option> </select> </div> <div> <input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result"> <label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox"> Send even when there aren't any new results </label> </div> <div class="action-panel-control-wrap option-text-in-email-wrap"> <label for="saved-search-email-text" class="action-panel-label"> Optional text in email: </label> <textarea name="saved-search-email-text" id="saved-search-email-text" class="saved-search-email-text"></textarea> </div> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Saving..." data-ga-category="save_share" data-ga-action="alert" data-ga-label="save"> Save </button> <button class="action-panel-cancel" aria-label="Close 'Your saved search' panel" ref="linksrc=close_saved_search_panel" aria-controls="saved-search-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="alert" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true"> <div class="inner-wrap"> <h2 class="action-panel-heading"> Create a file for external citation management software </h2> <form id="citation-manager-action-panel-form" class="action-panel-content action-form" action="/results-export-ids/" data-by-search-action="/results-export-search-data/" data-by-ids-action="/results-export-ids/" method="post" data-by-search-method="post" data-by-ids-method="post"> <input type="hidden" name="csrfmiddlewaretoken" value="6UE9QyRSrkKuEXMZCVs9T4Yd9SaEeMVWhCCQ4JIA6c5IN667p5FBKHC2S7nAAMBj"> <input name="results-format" type="hidden" value="pubmed"/> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Sending..." data-ga-category="save_share" data-ga-action="citation_manager" data-ga-label="save"> Create file </button> <button class="action-panel-cancel" aria-label="Close 'Send citations to citation manager' panel" ref="linksrc=close_citation_manager_panel" aria-controls="citation-manager-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="citation_manager" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true"> <div class="inner-wrap"> <h2 class="action-panel-heading"> Your RSS Feed </h2> <form id="rss-action-panel-form" class="rss-action-panel-form action-panel-content action-form" data-create-rss-feed-url="/create-rss-feed-url/" data-search-form-term-value=""> <input type="hidden" name="csrfmiddlewaretoken" value="6UE9QyRSrkKuEXMZCVs9T4Yd9SaEeMVWhCCQ4JIA6c5IN667p5FBKHC2S7nAAMBj"> <div class="action-panel-control-wrap"> <label for="rss-name" class="action-panel-label required-field-asterisk"> Name of RSS Feed: </label> <input maxlength="200" placeholder="Name" type="text" name="rss-name" id="rss-name" class="rss-name" value='' required pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"> </div> <div class="rss-limit-wrap"> <div class="action-panel-control-wrap action-panel-extra-margin-top"> <label for="rss-limit" class="action-panel-label"> Number of items displayed: </label> <select id="rss-limit" class="no-border-panel-selector rss-limit"> <option value="5">5</option> <option value="10">10</option> <option value="15" selected="selected">15</option> <option value="20">20</option> <option value="50">50</option> <option value="100">100</option> </select> </div> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Creating..." data-ga-category="save_share" data-ga-action="alert" data-ga-label="save"> Create RSS </button> <button class="action-panel-cancel" aria-label="Close 'Your RSS' panel" ref="linksrc=close_rss_panel" aria-controls="rss-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="alert" data-ga-label="cancel"> Cancel </button> </div> <div class="action-panel-control-wrap rss-link-copy-wrap"> <label for="rss-link" class="usa-sr-only">RSS Link</label> <input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link"> <button type="button" disabled class="rss-link-copy-button disabled" data-ga-category="save_share" data-ga-action="rss" data-ga-label="copy"> Copy </button> </div> </form> </div> </div> </div> </div> <div class="article-page" id="article-page" data-article-pmid="20346144"> <aside class="page-sidebar"> <div class="inner-wrap"> <div class="full-text-links"> <div class="full-view"> <h3 class="title"> Full text links </h3> <div class="full-text-links-list"> <a 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data-myncbi-max-collection-name-length="100" data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="6UE9QyRSrkKuEXMZCVs9T4Yd9SaEeMVWhCCQ4JIA6c5IN667p5FBKHC2S7nAAMBj"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio" id="collections-action-dialog-new" class="collections-new" name="collections" value="new" data-ga-category="collections_button" data-ga-action="click" data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio" id="collections-action-dialog-existing" class="collections-existing" name="collections" value="existing" checked="true" data-ga-category="collections_button" data-ga-action="click" data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk"> Name your collection: </label><input type="text" name="add-to-new-collection" id="collections-action-dialog-add-to-new" class="collections-action-add-to-new" pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/" maxlength="100" data-ga-category="collections_button" data-ga-action="create_collection" data-ga-label="non_favorties_collection"><div class="collections-new-name-too-long usa-input-error-message selection-validation-message"> Name must be less than 100 characters </div></div><div class="action-panel-control-wrap existing-collections-controls"><label for="collections-action-dialog-add-to-existing" class="action-panel-label"> Choose a collection: </label><select id="collections-action-dialog-add-to-existing" class="action-panel-selector collections-action-add-to-existing" data-ga-category="collections_button" data-ga-action="select_collection" data-ga-label="($('.collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'"></select><div class="collections-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your collection due to an error<br><a href="#">Please try again</a></div></div></div><div class="action-panel-actions"><button class="action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore data-ga-category="collections_button" data-ga-action="click" data-ga-label="add"> Add </button><button class="action-panel-cancel" aria-label="Close 'Add to Collections' panel" ref="linksrc=close_collections_panel" aria-controls="collections-action-panel" aria-expanded="false" data-ga-category="collections_button" data-ga-action="click" data-ga-label="cancel"> Cancel </button></div></form></div></div></div></div><button class="more-actions-button more-actions-dialog-trigger" title="Open dialog with more actions to take" ref="linksrc=more_actions_btn"></button><div class="display-options" 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href="/?term=McHugh+D&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Douglas McHugh">Douglas McHugh</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="The Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN 47405, USA." href="#full-view-affiliation-1" ref="linksrc=author_aff"> 1 </a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Hu+SS&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Sherry S J Hu">Sherry S J Hu</a><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Rimmerman+N&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Neta 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N-arachidonoyl glycine, an abundant endogenous lipid, potently drives directed cellular migration through GPR18, the putative abnormal cannabidiol receptor </h1> <div class="short-citation"> <span class="authors-list"> <span class="authors-list-item "><span class="full-name">Douglas McHugh</span><span class="citation-separator">&nbsp;et al.</span></span> </span> <span class="citation-journal"> BMC Neurosci<span class="citation-separator">.</span> </span> <span class="date"> <time class="citation-year" datetime="2010">2010</time><span class="citation-separator">.</span> </span> </div> <div class="more-article-details-section with-display-settings"> <button class="more-article-details" id="toggle-article-details" data-alt-text="Hide details" ref="linksrc=toggle_article_details" aria-controls="full-article-details" aria-expanded="false" data-fetch-url="/20346144/long-authors/" data-id-prefix=""> <span class="title">Show details</span> </button> <div class="display-options" 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data-ga-category="search" data-ga-action="author_link" data-ga-label="Douglas McHugh">Douglas McHugh</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="The Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN 47405, USA." href="#short-view-affiliation-1" ref="linksrc=author_aff"> 1 </a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Hu+SS&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Sherry S J Hu">Sherry S J Hu</a><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Rimmerman+N&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Neta Rimmerman">Neta Rimmerman</a><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Juknat+A&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Ana Juknat">Ana Juknat</a><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Vogel+Z&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Zvi Vogel">Zvi Vogel</a><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Walker+JM&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="J Michael Walker">J Michael Walker</a><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name" href="/?term=Bradshaw+HB&amp;cauthor_id=20346144" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Heather B Bradshaw">Heather B Bradshaw</a></span> </div> </div> <div class="affiliations"> <h3 class="title"> Affiliation </h3> <ul class="item-list"> <li data-affiliation-id="short-view-affiliation-1" id="short-view-affiliation-1" ><sup class="key">1</sup> The Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN 47405, USA.</li> </ul> </div> </div> <ul class="identifiers" id="short-view-identifiers"> <li> <span class="identifier pubmed"> <span class="id-label"> PMID: </span> <strong class="current-id" title="PubMed ID">20346144</strong> </span> </li> <li> <span class="identifier pmc"> <span class="id-label"> PMCID: </span> <a class="id-link" target="_blank" rel="noopener" ref="linksrc=article_id_link&amp;article_id=PMC2865488&amp;id_type=PMC" href="https://pmc.ncbi.nlm.nih.gov/articles/PMC2865488/" data-ga-category="full_text" data-ga-action="PMCID" > PMC2865488 </a> </span> </li> <li> <span class="identifier doi"> <span class="id-label"> 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class="abstract-content selected" id="eng-abstract"> <p> <strong class="sub-title"> Background: </strong> Microglia provide continuous immune surveillance of the CNS and upon activation rapidly change phenotype to express receptors that respond to chemoattractants during CNS damage or infection. These activated microglia undergo directed migration towards affected tissue. Importantly, the molecular species of chemoattractant encountered determines if microglia respond with pro- or anti-inflammatory behaviour, yet the signaling molecules that trigger migration remain poorly understood. The endogenous cannabinoid system regulates microglial migration via CB2 receptors and an as yet unidentified GPCR termed the 'abnormal cannabidiol' (Abn-CBD) receptor. Abn-CBD is a synthetic isomer of the phytocannabinoid cannabidiol (CBD) and is inactive at CB1 or CB2 receptors, but functions as a selective agonist at this Gi/o-coupled GPCR. N-arachidonoyl glycine (NAGly) is an endogenous metabolite of the endocannabinoid anandamide and acts as an efficacious agonist at GPR18. Here, we investigate the relationship between NAGly, Abn-CBD, the unidentified 'Abn-CBD' receptor, GPR18, and BV-2 microglial migration. </p> <p> <strong class="sub-title"> Results: </strong> Using Boyden chamber migration experiments, yellow tetrazolium (MTT) conversion, In-cell Western, qPCR and immunocytochemistry we show that NAGly, at sub-nanomolar concentrations, and Abn-CBD potently drive cellular migration in both BV-2 microglia and HEK293-GPR18 transfected cells, but neither induce migration in HEK-GPR55 or non-transfected HEK293 wildtype cells. Migration effects are blocked or attenuated in both systems by the 'Abn-CBD' receptor antagonist O-1918, and low efficacy agonists N-arachidonoyl-serine and cannabidiol. NAGly promotes proliferation and activation of MAP kinases in BV-2 microglia and HEK293-GPR18 cells at low nanomolar concentrations - cellular responses correlated with microglial migration. Additionally, BV-2 cells show GPR18 immunocytochemical staining and abundant GPR18 mRNA. qPCR demonstrates that primary microglia, likewise, express abundant amounts of GPR18 mRNA. </p> <p> <strong class="sub-title"> Conclusions: </strong> NAGly is the most effective lipid recruiter of BV-2 microglia currently reported and its effects mimic those of Abn-CBD. The data generated from this study supports the hypothesis that GPR18 is the previously unidentified 'Abn-CBD' receptor. The marked potency of NAGly acting on GPR18 to elicit directed migration, proliferation and perhaps other MAPK-dependent phenomena advances our understanding of the lipid-based signaling mechanisms employed by the CNS to actively recruit microglia to sites of interest. It offers a novel research avenue for developing therapeutics to elicit a self-renewing population of neuroregenerative microglia, or alternatively, to prevent the accumulation of misdirected, pro-inflammatory microglia which contribute to and exacerbate neurodegenerative disease. </p> </div> </div> <p class="disclaimer" id="disclaimer"> <a href="/disclaimer/" target="_blank" data-ga-category="literature_resources" data-ga-action="disclaimer_link" data-ga-label="under_abstract">PubMed Disclaimer</a> </p> <div class="figures" id="figures"> <h2 class="title"> Figures </h2> <div class="figures-list" id="slides-container" itemscope itemtype="http://schema.org/ImageGallery"> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="0" data-label-slug="figure-1"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/a0c8adeecedd/1471-2202-11-44-1.jpg" itemprop="contentUrl" aria-describedby="figure-caption-0" role="button" data-image-width="600" data-image-height="943" data-image-alt="Figure 1" data-pmc-id="PMC2865488" data-figure-id="F1"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-0" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/f1708847ff41/1471-2202-11-44-1.gif" alt="Figure 1" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="943"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 1 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced directed BV-2 microglial migration… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 1 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced directed BV-2 microglial migration </b> . (A) BV-2 microglial migration in response to…</div> </div> <figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 1 </strong> <div class="figure-caption-contents"><b>NAGly-induced directed BV-2 microglial migration</b>. (A) BV-2 microglial migration in response to basal conditions; vh (0.1% DMSO); 1 μM fMLP; 1 μM LPA; 0.1 nM - 300 μM NAGly. * = P &lt; 0.05, ** = P &lt; 0.01 compared to 1 μM fMLP; one-way ANOVA; <i>n </i>= 8. Insert is a filter photograph of one random field of view at ×40 magnification indicating the migration produced by 100 nM NAGly. The 10 μm diameter pores can be discerned as the clear unstained circles. (B) BV-2 microglial migration in response to basal conditions; vh (0.1% DMSO); 1 μM fMLP ± concentration gradient; 0.1 nM - 10 μM NAGly ± concentration gradient. ** = P &lt; 0.01 compared to the corresponding concentration gradient; Student's unpaired <i>t</i>-test; <i>n </i>= 3. Insert is a filter photograph of one random field of view at ×40 magnification indicating the migration produced by 100 nM NAGly in the absence of a concentration gradient.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="1" data-label-slug="figure-2"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/086c0347fa10/1471-2202-11-44-2.jpg" itemprop="contentUrl" aria-describedby="figure-caption-1" role="button" data-image-width="600" data-image-height="1012" data-image-alt="Figure 2" data-pmc-id="PMC2865488" data-figure-id="F2"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-1" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/87397d728d69/1471-2202-11-44-2.gif" alt="Figure 2" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="1012"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 2 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced BV-2 microglial migration is… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 2 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced BV-2 microglial migration is concentration- and structure-dependent </b> . (A) BV-2 microglial migration…</div> </div> <figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 2 </strong> <div class="figure-caption-contents"><b>NAGly-induced BV-2 microglial migration is concentration- and structure-dependent</b>. (A) BV-2 microglial migration in response to 0.1 nM - 10 μM concentrations of NAGly; AEA; 2-AG; PEA; PALGly; Abn-CBD; O-1602; LPI; <i>n </i>= 3. (B) Filter photographs of one random field of view at ×40 magnification indicating the migration produced by 10 μM concentrations of NAGly, O-1602, 2-AG, Abn-CBD, AEA and LPI.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="2" data-label-slug="figure-3"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/54bd3f3283e2/1471-2202-11-44-3.jpg" itemprop="contentUrl" aria-describedby="figure-caption-2" role="button" data-image-width="600" data-image-height="1559" data-image-alt="Figure 3" data-pmc-id="PMC2865488" data-figure-id="F3"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-2" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/5eb363b616c9/1471-2202-11-44-3.gif" alt="Figure 3" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="1559"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 3 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced BV-2 cell proliferation and… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 3 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced BV-2 cell proliferation and MAPK enzyme activation </b> . (A) BV-2 microglial proliferation…</div> </div> <figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 3 </strong> <div class="figure-caption-contents"><b>NAGly-induced BV-2 cell proliferation and MAPK enzyme activation</b>. (A) BV-2 microglial proliferation in response to 0.01 nM - 100 μM concentrations of NAGly; AEA; 2-AG; <i>n </i>= 3. (B) p44/42 MAPK activation in BV-2 microglia in response to vh (0.1% DMSO) for 3 hours; 10 nM - 10 μM NAGly for 3 hours; 10 μM Ionomycin for 5 min. ** = P &lt; 0.01 compared to vh; one-way ANOVA; <i>n </i>= 3. (C) p38 MAPK activation in BV-2 microglia in response to vh (0.1% DMSO) for 3 hours; 10 nM - 10 μM NAGly for 3 hours; 10 μM Ionomycin for 5 min. ** = P &lt; 0.01 compared to vh; one-way ANOVA; <i>n </i>= 3. (D) JNK MAPK activation in BV-2 microglia in response to vh (0.1% DMSO) for 3 hours; 10 nM - 10 μM NAGly for 3 hours; 10 μM Ionomycin for 5 min. ** = P &lt; 0.01 compared to vh; one-way ANOVA; <i>n </i>= 3.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="3" data-label-slug="figure-4"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/144de9cee26c/1471-2202-11-44-4.jpg" itemprop="contentUrl" aria-describedby="figure-caption-3" role="button" data-image-width="600" data-image-height="881" data-image-alt="Figure 4" data-pmc-id="PMC2865488" data-figure-id="F4"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-3" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/055ba570017e/1471-2202-11-44-4.gif" alt="Figure 4" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="881"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 4 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced BV-2 microglial migration is… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 4 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced BV-2 microglial migration is G <sub> i/o </sub> -receptor mediated and can be antagonized… </b></div> </div> <figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 4 </strong> <div class="figure-caption-contents"><b>NAGly-induced BV-2 microglial migration is G<sub>i/o</sub>-receptor mediated and can be antagonized</b>. (A) BV-2 microglial migration in response to basal conditions; vh (0.1% DMSO); 1 μM fMLP; 1 μM fMLP + 100 nM SR144528; 1 μM NAGly; 1 μM NAGly + 1 μM SR141716A; 1 μM NAGly + 100 nM SR141716A; 1 μM NAGly + 100 nM SR144528;. ** = P &lt; 0.01 compared to 1 μM NAGly; tt = P &lt; 0.01 compared to 1 μM fMLP; Student's unpaired <i>t</i>-test; <i>n </i>= 3. (B) BV-2 microglial migration in response to 0.1 nM - 10 μM NAGly ± 1 μM O-1918, ± 1 μM ARA-S, or ± 24 h pre-treatment with 1 μg/ml PTX; <i>n </i>= 3.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="4" data-label-slug="figure-5"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/db228c71f0bb/1471-2202-11-44-5.jpg" itemprop="contentUrl" aria-describedby="figure-caption-4" role="button" data-image-width="600" data-image-height="624" data-image-alt="Figure 5" data-pmc-id="PMC2865488" data-figure-id="F5"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-4" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/e3c7f4067954/1471-2202-11-44-5.gif" alt="Figure 5" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="624"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 5 </p> </strong> <div class="figure-caption-contents"><b> BV-2 microglia express GPR18 mRNA… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 5 </p> </strong> <div class="figure-caption-contents"><b> BV-2 microglia express GPR18 mRNA and GPR18 receptors </b> . (A) Gel electrophoresis of…</div> </div> <figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 5 </strong> <div class="figure-caption-contents"><b>BV-2 microglia express GPR18 mRNA and GPR18 receptors</b>. (A) Gel electrophoresis of BV-2 microglia and HEK293-GPR18 RT-qPCR products. RT-qPCR products were collected from the RT-qPCR run, loading buffer was added to the samples, and samples were run on a 2% agarose gel. No template control (NTC) and a control without reverse transcription (NRT) were used as controls. (B) Representative qPCR amplification curves showing the different amounts of mRNAs for GPR18 in primary microglia and BV-2 cells; <i>n </i>= 3.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="5" data-label-slug="figure-6"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/59f00a5824a1/1471-2202-11-44-6.jpg" itemprop="contentUrl" aria-describedby="figure-caption-5" role="button" data-image-width="600" data-image-height="457" data-image-alt="Figure 6" data-pmc-id="PMC2865488" data-figure-id="F6"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-5" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/0beb8c8243d5/1471-2202-11-44-6.gif" alt="Figure 6" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="457"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 6 </p> </strong> <div class="figure-caption-contents"><b> BV-2 microglia and HEK293-GPR18 transfected,… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 6 </p> </strong> <div class="figure-caption-contents"><b> BV-2 microglia and HEK293-GPR18 transfected, but not HEK293 wildtype, cells express GPR18 </b> .…</div> </div> <figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 6 </strong> <div class="figure-caption-contents"><b>BV-2 microglia and HEK293-GPR18 transfected, but not HEK293 wildtype, cells express GPR18</b>. Immunofluorescent confocal microscopy was conducted using an antibody against the GPR18 C-terminus (1:150; green), phalloidin to label actin (1:40; red), and DAPI (1.5 μg/ml) to label the nucleus (blue). <i>a</i>, HEK293 wildtype with DAPI and phalloidin. <i>b</i>, HEK293 wildtype with GPR18 antibody and phalloidin. <i>c</i>, HEK293 wildtype with GPR18 antibody, DAPI, and phalloidin. <i>d</i>, HEK293-GPR18 transfected with DAPI and phalloidin. <i>e</i>, HEK293-GPR18 transfected with GPR18 antibody and phalloidin. <i>f</i>, HEK293-GPR18 transfected with GPR18 antibody, DAPI, and phalloidin. <i>g</i>, BV-2 microglia with DAPI and phalloidin. <i>h</i>, BV-2 microglia with GPR18 antibody and phalloidin. <i>i</i>, BV-2 microglia with GPR18 antibody, DAPI, and phalloidin.</div> </figcaption> </figure> <figure class="figure-item tail" itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="6" data-label-slug="figure-7"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/17f94a8a5a86/1471-2202-11-44-7.jpg" itemprop="contentUrl" aria-describedby="figure-caption-6" role="button" data-image-width="600" data-image-height="600" data-image-alt="Figure 7" data-pmc-id="PMC2865488" data-figure-id="F7"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-6" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/63d9bc33f788/1471-2202-11-44-7.gif" alt="Figure 7" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="600"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 7 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced migration of HEK293-GPR18 and… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 7 </p> </strong> <div class="figure-caption-contents"><b> NAGly-induced migration of HEK293-GPR18 and HEK293-GPR55 cells </b> . (A) HEK293 wildtype and HEK293-GPR18…</div> </div> <figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 7 </strong> <div class="figure-caption-contents"><b>NAGly-induced migration of HEK293-GPR18 and HEK293-GPR55 cells</b>. (A) HEK293 wildtype and HEK293-GPR18 transfected cell migration in response to 0.1 nM - 10 μM NAGly; <i>n </i>= 3. (B) Filter photographs of one random field of view at ×40 magnification indicating the migration produced by 10 μM concentrations of NAGly in HEK293-GPR18 and HEK293 wildtype cells. The 10 μm diameter pores can be discerned as the clear unstained circles. (C) HEK293-GPR18 cell migration in response to Vh (0.1% DMSO); 1 μM NAGly; 1 μM NAGly ± 1 μM O-1918; 1 μM NAGly ± 1 μM ARA-S. *** = P &lt; 0.001 compared to 1 μM NAGly; one-way ANOVA; <i>n </i>= 3. HEK293-GPR18 cell migration in response to 1 μM Abn-CBD; 1 μM Abn-CBD ± 1 μM O-1918; 1 μM Abn-CBD ± 1 μM ARA-S; ttt = P &lt; 0.001 compared to 1 μM Abn-CBD; <i>n </i>= 3. HEK293-GPR18 cell migration in response to 1 μM O-1602; 1 μM O-1602 ± 1 μM O-1918; 1 μM O-1602 ± 1 μM ARA-S. §§§ = P &lt; 0.001 compared to 1 μM O-1602; one-way ANOVA; <i>n </i>= 3. (D) BV-2 microglia and HEK293-GPR18 cell migration in response to Vh (0.1% DMSO); 1 μM NAGly; 1 μM NAGly = 1 μM CBD; *** = P &lt; 0.001 compared to 1 μM NAGly; one-way ANOVA; <i>n </i>= 3. (E) p44/42 MAPK activation in HEK293-GPR18 cells in response to vh (0.1% DMSO) for 3 hours; 10 nM - 10 μM NAGly for 3 hours; 10 μM Ionomycin for 5 min. ** = P &lt; 0.01 compared to vh; one-way ANOVA; <i>n </i>= 3. (F) HEK293-GPR55 cell migration in response to 0.1 nM - 10 μM concentrations of LPI, Abn-CBD, NAGly, O-1602 and Vh (0.1% DMSO); <i>n </i>= 3.</div> </figcaption> </figure> <figure class="figure-item tail" itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="7" data-label-slug="figure-8"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/e70f94d5c6aa/1471-2202-11-44-8.jpg" itemprop="contentUrl" aria-describedby="figure-caption-7" role="button" data-image-width="600" data-image-height="393" data-image-alt="Figure 8" data-pmc-id="PMC2865488" data-figure-id="F8"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-7" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a50e/2865488/6bd9d1376aec/1471-2202-11-44-8.gif" alt="Figure 8" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="600"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="393"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 8 </p> </strong> <div class="figure-caption-contents"><b> GPR18 antibody recognizes hGPR18 receptors… </b></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 8 </p> </strong> <div class="figure-caption-contents"><b> GPR18 antibody recognizes hGPR18 receptors stably expressed in HEK293-GPR18 cells </b> . Immunofluorescent confocal…</div> </div> <figcaption id="figure-caption-7" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 8 </strong> <div class="figure-caption-contents"><b>GPR18 antibody recognizes hGPR18 receptors stably expressed in HEK293-GPR18 cells</b>. Immunofluorescent confocal microscopy was conducted using HEK293 cells stably transfected with HA11-tagged hGPR18 and HA11 (1:500) and GPR18 (1:500) antibodies. <i>a</i>, HA11 antibody (detected with Texas Red secondary; red) staining shows HA11-hGPR18 transfected cells. <i>b</i>, hGPR18 antibody (detected with FITC secondary; green) staining of the same cells identified with the HA11 antibody. <i>c</i>, HA11 antibody (detected with Texas Red secondary; red) staining shows HA11-hGPR18 transfected cells. <i>d</i>, hGPR18 antibody staining (detected with FITC secondary; green) was blocked when the GPR18 antibody was co-incubated with immunizing protein (30 μg/ml) for 1 hour before administration. DAPI (1.5 μg/ml; blue) was used to label the nucleus of all HEK293-GPR18 cells.</div> </figcaption> </figure> </div> <div class="see-all actions-bar"> <button class="show-more" aria-controls="slides-container" data-ga-category="figure" data-ga-action="show_all"> All figures (8) </button> </div> <!-- Root element of PhotoSwipe. Must have class pswp. --> <div class="pswp figures-dialog" id="figures-dialog" tabindex="-1" role="dialog" aria-hidden="true" data-image-gallery-uid="article-figures" aria-label="Figures Dialog"> <!-- Background of PhotoSwipe. It's a separate element as animating opacity is faster than rgba(). --> <div class="pswp__bg"></div> <!-- Slides wrapper with overflow:hidden. --> <div class="pswp__scroll-wrap"> <!-- Container that holds slides. PhotoSwipe keeps only 3 of them in the DOM to save memory. 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protein, human"> Search in MeSH </a></li><li><a class="add-to-search-link dropdown-block-link" ref="linksrc=add_to_search_link" role="button" data-search-term="&quot;GPR18 protein, human&quot;[nm]" data-ga-category="search" data-ga-action="add_to_search" data-ga-label="&quot;GPR18 protein, human&quot;[nm]" href="#"> Add to Search </a></li></ul></div></div></div></li><li><div class="keyword-actions dropdown-block"><button class="keyword-actions-trigger trigger keyword-link" aria-label="Toggle dropdown menu for keyword N-arachidonylglycine" tabindex="0" aria-controls="keyword-actions-substances-5" aria-expanded="false" data-pinger-ignore> N-arachidonylglycine </button><div id="keyword-actions-substances-5" class="keyword-actions-dropdown dropdown dropdown-container" hidden aria-label="Dropdown menu for keyword N-arachidonylglycine"><div class="title">Actions</div><div class="content"><ul class="keyword-actions-links"><li><a class="search-in-pubmed-link dropdown-block-link" 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