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<meta name="description" content="We report that eight heterozygous missense mutations in TUBB3, encoding the neuron-specific beta-tubulin isotype III, result in a spectrum of human nervous system disorders that we now call the TUBB3 syndromes. Each mutation causes the ocular motility disorder CFEOM3, whereas some also result in int …"><meta name="keywords" content="pmid:20074521, PMC3164117, doi:10.1016/j.cell.2009.12.011, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov&#x27;t, Research Support, U.S. Gov&#x27;t, Non-P.H.S., Max A Tischfield, Hagit N Baris, Elizabeth C Engle, Amino Acid Sequence, Animals, Axons / metabolism, Brain / embryology, Brain / metabolism, Cell Survival, Child, Developmental Disabilities, Female, Humans, Kinesins / metabolism, Male, Mice, Mice, Inbred C57BL, Microtubules / metabolism, Models, Molecular, Molecular Sequence Data, Mutation, Missense, Protein Transport, Tubulin / chemistry, Tubulin / genetics, Tubulin / metabolism*, PubMed Abstract, NIH, NLM, NCBI, National Institutes of Health, National Center for Biotechnology Information, National Library of Medicine, MEDLINE"><meta name="robots" content="index,nofollow,noarchive"><meta property="og:title" content="Human TUBB3 mutations perturb microtubule dynamics, kinesin interactions, and axon guidance - PubMed"><meta property="og:url" content="https://pubmed.ncbi.nlm.nih.gov/20074521/"><meta property="og:description" content="We report that eight heterozygous missense mutations in TUBB3, encoding the neuron-specific beta-tubulin isotype III, result in a spectrum of human nervous system disorders that we now call the TUBB3 syndromes. Each mutation causes the ocular motility disorder CFEOM3, whereas some also result in int …"><meta property="og:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:image:secure_url" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:type" content="website"><meta property="og:site_name" content="PubMed"><meta name="twitter:domain" content="pubmed.ncbi.nlm.nih.gov"><meta name="twitter:card" content="summary_large_image"><meta name="twitter:title" content="Human TUBB3 mutations perturb microtubule dynamics, kinesin interactions, and axon guidance - PubMed"><meta name="twitter:url" content="https://pubmed.ncbi.nlm.nih.gov/20074521/"><meta name="twitter:description" content="We report that eight heterozygous missense mutations in TUBB3, encoding the neuron-specific beta-tubulin isotype III, result in a spectrum of human nervous system disorders that we now call the TUBB3 syndromes. Each mutation causes the ocular motility disorder CFEOM3, whereas some also result in int …"><meta name="twitter:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg">
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<input type="email" aria-label="Recipient Email Address" placeholder="email@example.com" id="email-to" class="email-to" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="100" required>
</div>
<div class="action-panel-control-wrap">
<label for="email-from" class="action-panel-label">
From:
</label>
<input type="email" aria-label="Sender Email Address" placeholder="email@example.com" id="email-from" class="email-from" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="256">
</div>
<div class="action-panel-control-wrap">
<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
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="MxAE33XYOkMlYStGNDeFKKwHgjwGhU3c3kkJGTqkoVzPSjzwqWRHeS2lOneSavBw">
<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="MxAE33XYOkMlYStGNDeFKKwHgjwGhU3c3kkJGTqkoVzPSjzwqWRHeS2lOneSavBw">
<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="MxAE33XYOkMlYStGNDeFKKwHgjwGhU3c3kkJGTqkoVzPSjzwqWRHeS2lOneSavBw">
<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="MxAE33XYOkMlYStGNDeFKKwHgjwGhU3c3kkJGTqkoVzPSjzwqWRHeS2lOneSavBw">
<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="MxAE33XYOkMlYStGNDeFKKwHgjwGhU3c3kkJGTqkoVzPSjzwqWRHeS2lOneSavBw">
<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="20074521">
<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 class="link-item
dialog-focus"
href="https://linkinghub.elsevier.com/retrieve/pii/S0092-8674(09)01558-X"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3048&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=20074521&amp;db=pubmed&amp;nlmid=0413066"
title="See full text options at Elsevier Science"
data-ga-category="full_text"
data-ga-action="Elsevier Science"
data-ga-label="20074521"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--linkinghub.elsevier.com-ihub-images-celloa.png" alt="Elsevier Science full text link"><span class="text">
Elsevier Science
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/20074521/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=20074521&amp;db=pubmed&amp;nlmid=0413066"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="20074521"
><span class="text">
Free PMC article
</span></a>
</div>
</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/20074521/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/20074521/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="20074521" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F20074521%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-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-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="MxAE33XYOkMlYStGNDeFKKwHgjwGhU3c3kkJGTqkoVzPSjzwqWRHeS2lOneSavBw"><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"
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><sup class="key">1</sup> Department of Neurology, Children&#x27;s Hospital Boston, Boston, MA 02115, USA.</li>
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We report that eight heterozygous missense mutations in TUBB3, encoding the neuron-specific beta-tubulin isotype III, result in a spectrum of human nervous system disorders that we now call the TUBB3 syndromes. Each mutation causes the ocular motility disorder CFEOM3, whereas some also result in intellectual and behavioral impairments, facial paralysis, and/or later-onset axonal sensorimotor polyneuropathy. Neuroimaging reveals a spectrum of abnormalities including hypoplasia of oculomotor nerves and dysgenesis of the corpus callosum, anterior commissure, and corticospinal tracts. A knock-in disease mouse model reveals axon guidance defects without evidence of cortical cell migration abnormalities. We show that the disease-associated mutations can impair tubulin heterodimer formation in vitro, although folded mutant heterodimers can still polymerize into microtubules. Modeling each mutation in yeast tubulin demonstrates that all alter dynamic instability whereas a subset disrupts the interaction of microtubules with kinesin motors. These findings demonstrate that normal TUBB3 is required for axon guidance and maintenance in mammals.
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
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<p> Figure 1. Clinical spectrum and orbital imaging… </p>
</strong>
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</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 1. Clinical spectrum and orbital imaging of the TUBB3 syndromes </p>
</strong>
<div class="figure-caption-contents"><p> (AI) Study participant photographs.… </p></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. Clinical spectrum and orbital imaging of the TUBB3 syndromes
</strong>
<div class="figure-caption-contents">(AI) Study participant photographs. R262C can cause bilateral ptosis and severe CFEOM3 with the resting position of both eyes infraducted and abducted (A), moderate CFEOM3 that can be unilateral (B), and mild CFEOM3 (not shown). A similar spectrum is seen with D417N; severe CFEOM3 is shown in (E). A302T (C) and R380C (D) cause moderate to severe CFEOM3. Participants in (AE) have full facial movements. The axonal neuropathy in the participant with D417N (E) results in atrophy of the intrinsic foot muscles and a high arch (F). E410K (G) and R262H (H) result in severe CFEOM3 and facial weakness, and R262H also results in congenital ulnar deviation of the hand with joint contractures of the thumbs and fingers (I). (JL) MRI of the brainstem at the level of the oculomotor nerve (J) and orbital contents posterior to the globe (K) in a participant with predominantly left-sided CFEOM3 and a D417N substitution. Note unilateral hypoplasia of the left oculomotor nerve (J, arrow) and the atrophy of the levator palpebrae superioris (LPS), superior rectus (SR), and medial rectus (MR) muscles in (K). The inferior rectus (IR), lateral rectus (LR), and superior oblique (SO) muscles appear normal. (L) Control orbital MRI for comparison. (ON) denotes optic nerve.</div>
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<p> Figure 2. Spectrum of human brain malformations… </p>
</strong>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 2. Spectrum of human brain malformations correlate with specific <i> TUBB3 </i> mutations </p>
</strong>
<div class="figure-caption-contents"><p> (AG) Midline sagittal… </p></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. Spectrum of human brain malformations correlate with specific <i>TUBB3</i> mutations
</strong>
<div class="figure-caption-contents">(AG) Midline sagittal MRI showing the spectrum of corpus callosum (CC) dysgenesis; corresponding amino acid substitutions are noted to the left. R62Q (A) and most R262C (B) participants have normal CC development, whereas D417N subjects have hypoplasia of the posterior body (C, arrow). Subjects with A302T, E410K, and R262H have diffuse CC hypoplasia (DF). (G) Both R380C siblings have CC agenesis, and brainstem (arrow) and mild vermian hypoplasia (asterisk). (HN) Axial MRI from the same patient scans showing the spectrum of anterior commissure (AC) dysgenesis and overall loss of white matter compared to the normal R62Q scan (H, arrow indicates AC). (IL) Subjects have hypoplastic AC. R262H (M) and R380C (N) patient scans show AC agenesis and dysmorphic basal ganglia. The anterior limb of the left internal capsule is hypoplastic in R262H (M, arrow), while there is lack of clear separation between the caudate and putamen and bilateral hypoplasia of the anterior limbs of the internal capsule with R380C (N, arrows). (A-H) Anterior (A-D) and posterior (E-H) coronal sections showing the spectrum of basal ganglia dysmorphisms present in individuals with R262C, R262H, and R380C substitutions (Figure 3A-H). (A-D) Compared to a <i>TUBB3</i><sup>+/+</sup> scan (A), R262C reveals asymmetric basal ganglia with enlargement of the left caudate head and putamen (B arrow). (C) The twin of the R262H patient in (F, M) has dysgenesis of the left and right anterior limbs of the internal capsule (C arrows), apparent fusion of an enlarged left caudate head with the putamen, and dilatation of the left and right anterior horn of the lateral ventricle. (D) The older sibling of the patient scanned in (G, N) harboring an R380C substitution has hypoplasia of the anterior limb of the internal capsule (D arrow) and fusion of the left caudate head and underlying putamen. (E-H) Coronal MRI at the level of the caudate body and lateral ventricles is normal in (E). R262C and R262H subjects have hypoplasia of the left caudate body (F, G, arrows) and tail, and the R262H patient has dilatation of the left lateral ventricle. (H) The R380C patient has bilateral hypoplasia of the caudate body and tail, with Probst bundles of callosal axons that line the bodies of the lateral ventricles (arrow heads).</div>
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<strong class="figure-label">
<p> Figure 3. TUBB3 <sup> R262C/R262C </sup> mice have normal… </p>
</strong>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 3. TUBB3 <sup> R262C/R262C </sup> mice have normal cortical layering but show defects in axon guidance </p>
</strong>
<div class="figure-caption-contents"><p> (AD,… </p></div>
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<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. TUBB3<sup>R262C/R262C</sup> mice have normal cortical layering but show defects in axon guidance
</strong>
<div class="figure-caption-contents">(AD, n=5) WT and TUBB3<sup>R262C/R262C</sup> (EH, n=5) E18.5 coronal sections immunostained with markers specific for cortical layers show that the cortex has developed properly. Mild midline changes result from large Probst bundles (pb), comprised of stalled commissural axons adjacent to the midline. (I and K) Coronal sections from E18.5 WT (I, n=4) and TUBB3<sup>R262C/R262C</sup> (K, n=5) embryos show that the anterior commissure (red arrow) appears broken and fails to cross the midline, while the CC has crossed but is abnormally thick (red arrowhead) in the mutant. (J and L) E18.5 coronal sections from embryos immunostained with the axonal marker L1 show Probst bundles in a TUBB3<sup>R262C/R262C</sup> (L, arrowheads) mutant compared to WT (J). (MP) Whole-mount neurofilament staining of E11.5-12 WT (M, N; n=13) and TUBB3<sup>R262C/R262C</sup> (O, P; n=6) embryos. Mutant oculomotor (III) and trochlear (IV) nerves, as well the maxillary (Vm) and ophthalmic (Vo) divisions of the trigeminal nerve are stalled at E11.5 (O) compared to WT (M). At E12, the mutant oculomotor nerve follows an aberrant course adjacent to the trochlear nerve (P) compared to WT (N, ). CC = corpus callosum; e = eye; asterisk (*)= distal tip of oculomotor nerve.</div>
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<p> Figure 4. TUBB3 <sup> R262C/R262C </sup> mice have low… </p>
</strong>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 4. TUBB3 <sup> R262C/R262C </sup> mice have low TUBB3 protein levels, altered microtubule stability, and decreased… </p>
</strong>
</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. TUBB3<sup>R262C/R262C</sup> mice have low TUBB3 protein levels, altered microtubule stability, and decreased Kif21a interactions
</strong>
<div class="figure-caption-contents">(A) TUBB3 protein levels are reduced in TUBB3<sup>+/R262C</sup> (WT/KI) and TUBB3<sup>R262C/R262C</sup> (KI/KI) vs. WT mice. (B) TUBB3 R262C heterodimers incorporate into microtubules throughout cell bodies, neurites, and growth cones of dissociated cortical neurons as seen by co-localization with tyrosinated -tubulin and actin. Variable reductions in TUBB3 staining intensity are noted between WT and mutant neurons. (C) Levels of de-tyrosinated -tubulin are increased in brain lysates from mutant versus WT mice. (D) Brain lysates from TUBB3<sup>R262C/R262C</sup> mice show increased microtubule polymerization at steady-state levels despite lower levels of β-tubulin. Mutant TUBB3 is detected in the pellets (p). (E) Levels of Kif21a are reduced on TUBB3<sup>R262C/R262C</sup> mutant microtubules polymerized <i>in vitro</i> from brain lysates and incubated with ATP, whereas levels of Kif3a remain constant. (F) <i>In vitro</i> transcription and translation of WT and TUBB3 mutant heterodimers in rabbit reticulocyte lysate. Products analyzed by SDS (top) and non-denaturing (native, bottom) gel electrophoresis and stained with an anti-V5 antibody against the C-terminal tag demonstrate that although transcription and translation are not affected by the mutations, there can be significant and variable decreases in the yield of native heterodimers. (G) Synthesized WT, R262H, E410K, and D417H/N heterodimers cycle with native bovine tubulin at equivalent efficiency; vertical lines denote removed empty lanes. *<i>P</i>&lt;0.05, **<i>P</i>&lt;0.001.</div>
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<p> Figure 5. TUBB3 amino acid substitutions and… </p>
</strong>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 5. TUBB3 amino acid substitutions and phenotype-genotype-function correlations </p>
</strong>
<div class="figure-caption-contents"><p> (A) Schematic of the TUBB3 protein;… </p></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. TUBB3 amino acid substitutions and phenotype-genotype-function correlations
</strong>
<div class="figure-caption-contents">(A) Schematic of the TUBB3 protein; arrows indicate the location of each TUBB3 amino acid substitution. (B) Structure of the αβ-tubulin heterodimer from a rotated side view (pdb: 1JFF). α-tubulin is dark-shaded on right, and the arrows on the left point to each mutant residue, depicted in black, on β-tubulin. (C) Outside view of β-tubulin depicting each mutant residue in black with the exception of R62Q, which cannot be seen. The area proposed for motor protein interactions is depicted as a shaded grey oval. Residues R380, E410 and D417 are found in helices H11 and H12, respectively, on the external surface of β-tubulin. R262, found in the loop between helix H8 and strand 7, and A302, found in the loop following helix H9, are positioned laterally and below to helices H12 and H11, respectively. (D) Side interior view of β-tubulin showing the location of each mutant residue in black, with the exception of R380C, which is occluded by D417. R62 is positioned in the H1-S2 loop (N-loop) that forms lateral contacts with the M-loop of adjacent protofilaments. (E, top) Magnified image of β-tubulin depicting the putative hydrogen bond between the arginine side chain of R262 and the carbonyl oxygen of D417, both of which are shown in a stick representation. (E, middle and bottom) Both the R262C and R262H substitutions are predicted to break the hydrogen bond. (F) Phenotype-genotype and summary of functional data. AA = amino acid substitution; # ped = number of pedigrees; # aff = number of affected individuals; AC = anterior commissure hypoplasia; CC = corpus callosum hypoplasia; BG = basal ganglia dysgenesis; DD = developmental delay; FW = facial weakness; PN = progressive axonal sensorimotor polyneuropathy; CJC = congenital joint contractures; HD = heterodimer formation; HeLa = HeLa cell incorporation; MT dyn = microtubule dynamics (HS = highly stable, S = stable); Kip3p/2p = Kip3p and Kip2p microtubule plus-end accumulation ( ↓ decreased, ↑ increased). + denotes present; - denotes absent; /+ denotes that only a subset of subjects have the feature and/or the findings are mild; variable () denotes rare participants with the substitution without CFEOM3.</div>
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<p> Figure 6. TUBB3 disease amino acid substitutions… </p>
</strong>
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<strong class="figure-label">
<p> Figure 6. TUBB3 disease amino acid substitutions results in changes to microtubule dynamic instability </p>
</strong>
<div class="figure-caption-contents"><p> (A,… </p></div>
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Figure 6. TUBB3 disease amino acid substitutions results in changes to microtubule dynamic instability
</strong>
<div class="figure-caption-contents">(A, B) 4-D life-time plots depicting the lengths of microtubules (Y axis) over time (X axis) in G1 cells from (A) haploid WT and <i>TUB2</i> mutants and (B) heterozygous diploid WT and <i>TUB2</i> mutants demonstrate that Tub2p substitutions perturb microtubule dynamic instability. For WT and each mutation, one microtubule representing data from the collective analysis has been selected and plotted. (C) Summary table of individual dynamic instability parameters. Number of events is listed in parentheses. *<i>P</i>&lt;0.05, **<i>P</i>&lt;0.001, ***<i>P</i>&lt;0.0001.</div>
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<p> Figure 7. Kip3p and Kip2p levels are reduced at the plus-ends of mutant microtubules </p>
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Figure 7. Kip3p and Kip2p levels are reduced at the plus-ends of mutant microtubules
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<div class="figure-caption-contents">Merged z-stack images showing the levels of Kip3p-3YFP (AE, red) and Kip2p-3YFP (KO, red) on WT and mutant microtubules labeled with CFP-Tub1p (α-tubulin, green) in budding yeast. Corresponding Kip3p (FJ) and Kip2p (PT) YFP channels with signal intensities adjusted equally for WT and each <i>TUB2</i> mutant are provided for comparison. In WT cells, Kip3p-3YFP (A, F) and Kip2p-3YFP (K, P) are speckled along the length and accumulate at the plus-ends of growing microtubules labeled by white arrows. In contrast, mutant R262C (B), R262H (C), E410K (D), and D417N (E) cells all have a significant reduction of Kip3p-3YFP along the length and plus-ends of microtubules (white-arrows), and Kip2p-3YFP is speckled along the length but reduced or absent on microtubule plus-ends (white-arrows) in R262C (L), R262H (M), E410K (N), and D417H (O) cells. Graphs depicting the overall mean levels of Kip3p-YFP (U) and Kip2p-3YFP (V) on microtubule plus-ends in WT and mutant cells. *<i>P</i>&lt;0.05, ** <i>P</i>&lt;0.001, *** <i>P</i>&lt;0.0001.</div>
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