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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="KCUyYnaV4TnqyU8fy5hI3hTPUZCna9A81pEDBdDhEuaUsle5boUKxppts3kz3K8s">
<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="KCUyYnaV4TnqyU8fy5hI3hTPUZCna9A81pEDBdDhEuaUsle5boUKxppts3kz3K8s">
<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="KCUyYnaV4TnqyU8fy5hI3hTPUZCna9A81pEDBdDhEuaUsle5boUKxppts3kz3K8s">
<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="KCUyYnaV4TnqyU8fy5hI3hTPUZCna9A81pEDBdDhEuaUsle5boUKxppts3kz3K8s">
<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="KCUyYnaV4TnqyU8fy5hI3hTPUZCna9A81pEDBdDhEuaUsle5boUKxppts3kz3K8s">
<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="17567819">
<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="http://www.jneurosci.org/cgi/pmidlookup?view=long&amp;pmid=17567819"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3051&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=17567819&amp;db=pubmed&amp;nlmid=8102140"
title="See full text options at HighWire"
data-ga-category="full_text"
data-ga-action="HighWire"
data-ga-label="17567819"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--highwire.stanford.edu-icons-externalservices-pubmed-standard-jneuro_final_free.gif" alt="HighWire full text link"><span class="text">
HighWire
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/17567819/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=17567819&amp;db=pubmed&amp;nlmid=8102140"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="17567819"
><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="/17567819/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/17567819/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="17567819" 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%2F17567819%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="KCUyYnaV4TnqyU8fy5hI3hTPUZCna9A81pEDBdDhEuaUsle5boUKxppts3kz3K8s"><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"
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ClC-2 is a broadly expressed plasma membrane chloride channel that is modulated by voltage, cell swelling, and pH. A human mutation leading to a heterozygous loss of ClC-2 has previously been reported to be associated with epilepsy, whereas the disruption of Clcn2 in mice led to testicular and retinal degeneration. We now show that the white matter of the brain and spinal cord of ClC-2 knock-out mice developed widespread vacuolation that progressed with age. Fluid-filled spaces appeared between myelin sheaths of the central but not the peripheral nervous system. Neuronal morphology, in contrast, seemed normal. Except for the previously reported blindness, neurological deficits were mild and included a decreased conduction velocity in neurons of the central auditory pathway. The heterozygous loss of ClC-2 had no detectable functional or morphological consequences. Neither heterozygous nor homozygous ClC-2 knock-out mice had lowered seizure thresholds. Sequencing of a large collection of human DNA and electrophysiological analysis showed that several ClC-2 sequence abnormalities previously found in patients with epilepsy most likely represent innocuous polymorphisms.
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<p> Figure 1. </p>
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<div class="figure-caption-contents"><p> Spongiform vacuolation in white matter… </p></div>
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<p> Figure 1. </p>
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<div class="figure-caption-contents"><p> Spongiform vacuolation in white matter tracts of ClC-2 KO mice. <b> <i> A </i> </b> , Hematoxylineosin-stained… </p></div>
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Figure 1.
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<div class="figure-caption-contents">Spongiform vacuolation in white matter tracts of ClC-2 KO mice. <b><i>A</i></b>, Hematoxylineosin-stained brain section of a 22-month-old <i>Clcn2</i>
<sup>/</sup> mouse shows vacuolation in white matter tracts of the cerebellum (a), corpus callosum (b), internal capsule (c), and brainstem (d). <b><i>B</i></b>, An age-matched WT brain shows much less vacuolation, which is mostly attributable to capillaries. <b><i>CF</i></b>, Progressive vacuolation in <i>Clcn2</i>
<sup>/</sup> cerebellum from P14 (<b><i>C</i></b>) (no vacuolation) to P28 (<b><i>D</i></b>), 4 months (<b><i>E</i></b>), and 22 months (<b><i>F</i></b>). <b><i>G</i></b>, Semithin sections of the middle cerebellar peduncle of 2-, 5-, and 14-month-old WT and ClC-2 KO mice used to quantify the area of vacuolation. Abundant vacuoles (asterisks) were visible in cerebellar white matter of 5- and 14-month-old KO but not WT animals, in which capillaries (c) are indicated. <b><i>H</i></b>, The white area of randomly taken sections was determined by automated image analysis and shown as the percentage of the total area [5.2 ± 0.6% (KO) vs 1.4 ± 1.3% (WT) at 2 months, 17.8 ± 0.9% (KO) vs 2.1 ± 0.5% (WT) at 5 months, and 21.3 ± 1.2% (KO) vs 3.3 ± 1.2% (WT) at 14 months; 10 sections per mouse, 3 mice each]. Error bars indicate SEM. <b><i>I</i></b>, Prominent vacuolation in the white matter of the spinal cord of a 14-month-old KO but not WT mouse. Scale bars: <b><i>A</i></b>, <b><i>B</i></b>, 1 mm; <b><i>CF</i></b>, 0.25 mm; <b><i>G</i></b>, 20 μm; <b><i>I</i></b>, 0.5 mm.</div>
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<p> Figure 2. </p>
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<div class="figure-caption-contents"><p> Ultrastructural analysis of vacuoles and… </p></div>
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<p> Figure 2. </p>
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<div class="figure-caption-contents"><p> Ultrastructural analysis of vacuoles and myelin. <b> <i> A </i> </b> , Semithin sections of the sciatic… </p></div>
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Figure 2.
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<div class="figure-caption-contents">Ultrastructural analysis of vacuoles and myelin. <b><i>A</i></b>, Semithin sections of the sciatic nerve show no difference between the genotypes at 14 months of age (14m). <b><i>B</i></b>, Western blot analysis detects the ClC-2 protein in brain but not in sciatic nerve (sciatic n.). KO lysates showed the specificity of the ClC-2 antibody. Actin served as a loading control. <b><i>CE</i></b>, Electron micrographs showing typical vacuoles in white matter tracts of ClC-2 KO cerebellum. These vacuoles (asterisks) contained aberrant myelin sheets (arrows) and were surrounded by thin myelin sheaths (arrowheads). a, Axon. <b><i>D</i></b>, Myelinated axon surrounded by a vacuole and a normal-appearing oligodendrocyte (n). <b><i>F</i></b>, Part of WT white matter cerebellum. <b><i>G</i></b>, The structure of myelin sheaths in the area of best compaction revealed no difference in myelin compaction. Distances between MDLs were unchanged in the KO [10.7 ± 0.1 nm (WT) vs 10.8 ± 0.1 nm (KO); <i>n</i> &lt; 90, 3 animals each]. 7w, 7 weeks old. <b><i>H</i></b>, Semithin sections of the optic nerve of 7-week-old (7w) WT and <i>Clcn2</i>
<sup>/</sup> mice revealed no vacuolation in the KO (asterisks indicate capillary). Higher magnification is shown in supplemental Fig. S3 (available at <ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://www.jneurosci.org">www.jneurosci.org</ext-link> as supplemental material). Scale bars: <b><i>A</i></b>, 20 μm; <b><i>CF</i></b>, 1 μm; <b><i>G</i></b>, 20 nm; <b><i>H</i></b>, 10 μm.</div>
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<p> Figure 3. </p>
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<div class="figure-caption-contents"><p> Inflammation and impaired bloodbrain barrier… </p></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
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<p> Figure 3. </p>
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<div class="figure-caption-contents"><p> Inflammation and impaired bloodbrain barrier in ClC-2 KO mice and functional tests of… </p></div>
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Figure 3.
</strong>
<div class="figure-caption-contents">Inflammation and impaired bloodbrain barrier in ClC-2 KO mice and functional tests of motor coordination and central nerve conductance. <b><i>A</i></b>, Quantitative PCR (qPCR) analysis of selected transcripts from cerebella of 5-week-old (<i>n</i> = 6, each genotype) and 24-week-old (<i>n</i> = 3, each genotype) animals. Expression levels are given as a percentage of wild type. Genes analyzed by qPCR are as follows: 1, cathepsin-S (accession #NM021281); 2, osteopontin (NM009263); 3, lysozyme (NM013590); 4, F4/80 (NM010130); 5, hexosaminidase (NM010422); 6, leukocyte-derived chemotaxin (NM010701); 7, complement component 1 (NM007572); 8, glutathion peroxidase (NM008161); 9, GFAP (NM010277). Except for GFAP and F4/80, these genes were also found to be upregulated in expression profiling (supplemental Table 1, available at <ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://www.jneurosci.org">www.jneurosci.org</ext-link> as supplemental material). Rel., Relative. <b><i>B</i></b>, <b><i>C</i></b>, Red staining by labeled GSA showed microglia activation in 8-month-old <i>Clcn2</i>
<sup>/</sup> (<b><i>C</i></b>) but not WT (<b><i>B</i></b>) cerebella. <b><i>D</i></b>, <b><i>E</i></b>, Diaminobenzidine staining (brown) for HRP that was previously injected intravenously into 8-month-old (8m) <i>Clcn2</i>
<sup>/</sup> (<b><i>E</i></b>) or WT (<b><i>D</i></b>) mice revealed extravasation (arrows), indicative of a disrupted bloodbrain barrier only in the KO. Nuclei were counterstained with methylgreen. <b><i>F</i></b>, In rotarod tests, ClC-2 KO mice (<i>n</i> = 5) performed as well as WT (<i>n</i> = 4) mice. Motor skills improved with the number of trials in both groups. <b><i>G</i></b>, IPIs between waves I and III of auditory brain stem responses were significantly increased in older animals, indicating a slowed nerve conductance in the KO [5 weeks (5w), ***<i>p</i> &lt; 0.1%; 12 weeks (12w), **<i>p</i> &lt; 1%; <i>n</i> = 518 mice, same as in supplemental Fig. S7, available at <ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://www.jneurosci.org">www.jneurosci.org</ext-link> as supplemental material]. 3w, 3 weeks. Error bars indicate SEM. Scale bars: <b><i>B</i></b>, <b><i>C</i></b>, 0.05 mm; <b><i>D</i></b>, <b><i>E</i></b>, 0.1 mm.</div>
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<div class="figure-caption-contents"><p> Subcellular localization of ClC-2 in… </p></div>
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<p> Figure 4. </p>
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<div class="figure-caption-contents"><p> Subcellular localization of ClC-2 in brain. Confocal images of brain sections of 5-week-old… </p></div>
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Figure 4.
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<div class="figure-caption-contents">Subcellular localization of ClC-2 in brain. Confocal images of brain sections of 5-week-old WT and ClC-2 KO mice double stained for ClC-2 (green) and marker proteins (red) including GFAP (<b><i>AC</i></b>), NeuN (<b><i>D</i></b>), CC1 (<b><i>E</i></b>), and Cx47 (<b><i>F</i></b>). <b><i>A</i></b>, In the hippocampus, ClC-2 colocalized with GFAP in astrocytic endfeet (arrows) surrounding blood vessels (asterisks). <b><i>B</i></b>, <b><i>C</i></b>, In the cerebellum, ClC-2 was found in GFAP-positive Bergman glia of the molecular layer (<b><i>B</i></b>) and in myelinated fiber tracts (<b><i>C</i></b>). <b><i>C</i></b>, <b><i>D</i></b>, In the latter region, ClC-2 did not significantly overlap with either GFAP (<b><i>C</i></b>) or NeuN (<b><i>D</i></b>). <b><i>E</i></b>, Cells with numerous ClC-2 puncta around their cell bodies were identified as oligodendrocytes by staining for CC1. <b><i>F</i></b>, Many of these ClC-2-positive puncta also stained for Cx47. Scale bars, 20 μm. Blue indicates TOTO staining of nuclei.</div>
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<p> Figure 5. </p>
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<div class="figure-caption-contents"><p> Potential role of ClC-2 in… </p></div>
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<p> Figure 5. </p>
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<div class="figure-caption-contents"><p> Potential role of ClC-2 in epilepsy. <b> <i> A </i> </b> , Two-electrode voltage-clamp analysis of <i> Xenopus… </i> </p></div>
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Figure 5.
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<div class="figure-caption-contents">Potential role of ClC-2 in epilepsy. <b><i>A</i></b>, Two-electrode voltage-clamp analysis of <i>Xenopus</i> oocytes previously injected with 10 ng of WT ClC-2 cRNA (■) or with a construct modeled on the truncating Gins597 mutation (○) from a family with epilepsy (Haug et al., 2003). Injecting half the amount (5 ng) of WT ClC-2 cRNA gave about half the current amplitude (□). Coinjection of 5 ng of WT and 5 ng of Gins597 cRNA (•) yielded similar current amplitudes, indicating that the mutant lacks a dominant-negative effect. Currents were obtained from &gt;15 oocytes from three batches, averaged, and normalized to wild type. <b><i>B</i></b>, Two-electrode voltage-clamp analysis of two sequence variants identified in epileptic patients (D'Agostino et al., 2004) in the present screen of patients with leukodystrophy and by Stogmann et al. (2006). Currents from ClC-2_E718D (▾; <i>n</i> = 21 oocytes) and ClC-2_R688Q (○; <i>n</i> = 23) were indistinguishable from WT ClC-2 currents (■; <i>n</i> = 19). Averaged and normalized currents from three batches of oocytes are shown. <b><i>C</i></b>, <b><i>D</i></b>, Seizure susceptibility of WT, <i>Clcn2</i>
<sup>+/</sup> (HET), and <i>Clcn2</i>
<sup>/</sup> (KO) mice as determined by exposure to fluorethyl (applied at 10 ml/min to the chamber; <b><i>C</i></b>) or pentylenetetrazol (PTZ; injected at 50 mg/kg body weight; <b><i>D</i></b>). <b><i>C</i></b>, The apparent slight decrease in threshold in <i>Clcn2</i>
<sup>/</sup> mice (2.34 ± 0.13 min; <i>n</i> = 8 mice) was not statistically different (<i>p</i> = 0.072) from those of WT (2.95 ± 0.25; <i>n</i> = 12) or <i>Clcn2</i>
<sup>+/</sup> mice (2.91 ± 0.21 min; <i>n</i> = 3). <b><i>D</i></b>, There was no difference between seizure thresholds of WT and KO animals on exposure to PTZ (3.70 ± 1.41 vs 4.04 ± 1.25 min; <i>n</i> = 6 mice each). Error bars indicate SEM.</div>
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Related information
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Gene
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OMIM (cited)
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