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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="BC0p7yxliQQSYA0uhvK2Xyokyq7NTOiESpKuKo0HSrDmS16kUOn4rGUY6uPZMpQY">
<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="BC0p7yxliQQSYA0uhvK2Xyokyq7NTOiESpKuKo0HSrDmS16kUOn4rGUY6uPZMpQY">
<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="BC0p7yxliQQSYA0uhvK2Xyokyq7NTOiESpKuKo0HSrDmS16kUOn4rGUY6uPZMpQY">
<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="BC0p7yxliQQSYA0uhvK2Xyokyq7NTOiESpKuKo0HSrDmS16kUOn4rGUY6uPZMpQY">
<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="BC0p7yxliQQSYA0uhvK2Xyokyq7NTOiESpKuKo0HSrDmS16kUOn4rGUY6uPZMpQY">
<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="12805295">
<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=12805295"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3051&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=12805295&amp;db=pubmed&amp;nlmid=8102140"
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pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/12805295/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=12805295&amp;db=pubmed&amp;nlmid=8102140"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
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Free PMC article
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</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="/12805295/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/12805295/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="12805295" 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%2F12805295%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="BC0p7yxliQQSYA0uhvK2Xyokyq7NTOiESpKuKo0HSrDmS16kUOn4rGUY6uPZMpQY"><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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><sup class="key">1</sup> Institut für Genetik, Universität Bonn, D-53117 Bonn, Germany.</li>
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<p>
To further characterize the recently described gap junction gene connexin 47 (Cx47), we generated Cx47-null mice by replacing the Cx47 coding DNA with an enhanced green fluorescent protein (EGFP) reporter gene, which was thus placed under control of the endogenous Cx47 promoter. Homozygous mutant mice were fertile and showed no obvious morphological or behavioral abnormalities. Colocalization of EGFP fluorescence and immunofluorescence of cell marker proteins revealed that Cx47 was mainly expressed in oligodendrocytes in highly myelinated CNS tissues and in few calcium-binding protein S100beta subunit-positive cells but not in neurons or peripheral sciatic nerve. This corrects our previous conclusion that Cx47 mRNA is expressed in brain and spinal cord neurons (Teubner et al., 2001). Cx47 protein was detected by Western blot analysis after immunoprecipitation in CNS tissues of wild-type mice but not in heart or Cx47-deficient tissues. Electron microscopic analysis of CNS white matter in Cx47-deficient mice revealed a conspicuous vacuolation of nerve fibers, particularly at the site of the optic nerve where axons are first contacted by oligodendrocytes and myelination starts. Initial analyses of Cx32/Cx47-double-deficient mice showed that these mice developed an action tremor and died on average at 51 d after birth. The central white matter of these double-deficient mice exhibited much more abundant vacuolation in nerve fibers than mice deficient only in Cx47.
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<a class="figure-link"
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itemprop="contentUrl"
aria-describedby="figure-caption-0"
role="button"
data-image-width="720"
data-image-height="658"
data-image-alt="Figure 1."
data-pmc-id="PMC6740816"
data-figure-id="FIG1">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/695f61484e89/ns1137669001.gif"
alt="Figure 1." />
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<strong class="figure-label">
<p> Figure 1. </p>
</strong>
<div class="figure-caption-contents"><p> Generation of Cx47-deficient mice with… </p></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"><p> Generation of Cx47-deficient mice with the EGFP reporter gene (Cx47 <sup> EGFP </sup> ). <i> a… </i> </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.
</strong>
<div class="figure-caption-contents">Generation of Cx47-deficient mice with the EGFP reporter gene (Cx47 <sup>EGFP</sup>). <i>a</i>, Targeting scheme. WT, Wild-type allele; KO, knock-out allele. The coding region of the wild-type Cx47 gene (black arrow) is located on one exon and includes 1314 bp. After electrotrans-fecting HM-1 cells with the linearized targeting vector (KO-vektor), homologous recombination occurred in 10% of all clones as determined by PCR using the external sense primer (P2) and the EGFP-specific antisense primer (P4). In the mutated Cx47 allele, the coding region of Cx47 after the first seven codons was replaced by the EGFP cDNA and the PGK-HPRT minigene (gray arrows). The translational start codon of EGFP was cloned in-frame with the Cx47 coding region. The PGK-HPRT minigene was transcribed in the opposite direction. The backbone of the targeting vector consisted of pBlueskript SK II(+) (pBSK II; Stratagene). The 5 homologous region (5hom) spanned 2 kb; the 3 homologous region (3hom) included 5.5 kb. <i>b</i>, Southern blot analysis was performed to confirm homologous recombination. <i>Kpn</i>I-digested mouse kidney DNA was probed with a 1 kb external <i>Eco</i>RV<i>Kpn</i>I fragment (probe). The 7.0 kb band was derived from the wild-type allele; the larger 12.2 kb band resulted from the knock-out allele. <i>c</i>, Diagnostic PCR of tail-tip DNA was performed to screen pedigree mice for occurrence of wild-type and knock-out Cx47 <sup>EGFP</sup> alleles. A multiplex PCR was established using a Cx47 intron-specific sense primer (P1) and a Cx47 exon-specific antisense primer (P3), producing a 530 bp wild-type amplicon and an EGFP-specific antisense primer (P4), resulting in a 340 bp amplicon of the knock-out allele.</div>
</figcaption>
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itemprop="contentUrl"
aria-describedby="figure-caption-1"
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data-image-width="775"
data-image-height="1199"
data-image-alt="Figure 2."
data-pmc-id="PMC6740816"
data-figure-id="FIG2">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-1"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/6ca732337375/ns1137669002.gif"
alt="Figure 2." />
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<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 2. </p>
</strong>
<div class="figure-caption-contents"><p> Northern blot analysis of Cx47… </p></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"><p> Northern blot analysis of Cx47 expression ( <i> a </i> ) and EGFP expression (… </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.
</strong>
<div class="figure-caption-contents">Northern blot analysis of Cx47 expression (<i>a</i>) and EGFP expression (<i>b</i>) in adult mouse tissues. Equal amounts of total RNA (20 μg) from wild-type (+/+), heterozygous (+/-), and homozygous (-/-) Cx47 <sup>EGFP</sup> mice (littermates) were applied as demonstrated by staining of 28S and 18S ribosomal RNA with ethidium bromide (top). The blotted RNA (bottom) was probed with a 1.6 kb <i>Xba</i>I fragment containing part of the Cx47 coding region and 3-untranslated region (<i>a</i>) and a 1.0 kb <i>Sal</i>I<i>Afl</i>II fragment of the pEGFP-1 vector containing the EGFP-coding region and the SV40 poly(A) signal (b). The size of the corresponding transcripts is indicated in kilobases on the left.</div>
</figcaption>
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<figure class="figure-item "
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data-image-width="664"
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data-image-alt="Figure 3."
data-pmc-id="PMC6740816"
data-figure-id="FIG3">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-2"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/6cab699e5687/ns1137669003.gif"
alt="Figure 3." />
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<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"><p> Western blot analysis of protein… </p></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"><p> Western blot analysis of protein lysates from different HeLa connexin transfectants ( <i> a… </i> </p></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">Western blot analysis of protein lysates from different HeLa connexin transfectants (<i>a</i>) and after immunoprecipitation of protein lysates from Cx47 wild-type (+/+) and Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mouse tissues (<i>b</i>). Western blot analysis with Cx47 antibodies detected by <sup>125</sup>I-labeled protein A showed specific bands, at 50 kDa in HeLa Cx47 lysates and 75 kDa in HeLaCx47-EGFPlysates. Nonspecific bands at 30 and 100 kDa also appeared in control lysates of HeLa wild-type cells and Cx45 transfectants (<i>a</i>). Molecular masses are indicated in kilodaltons on the left. By Western blot analysis (ECL) with biotinylated Cx47 antibodies after immunoprecipitation with Cx47 antibodies in lysates of mouse tissue (<i>b</i>), Cx47 was found in the cerebellum and spinal cord of wild-type mice; in cerebrum, the signal was very weak. In heart (control tissue) and lysates from Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> littermates, no bands were detected. Numbers under each lane represent micrograms of total protein per microliter of lysate of the tissue indicated.</div>
</figcaption>
</figure>
<figure class="figure-item "
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href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/3dbc6a1d4bc7/ns1137669004.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="567"
data-image-height="853"
data-image-alt="Figure 4."
data-pmc-id="PMC6740816"
data-figure-id="FIG4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/f967fe8a74be/ns1137669004.gif"
alt="Figure 4." />
</a>
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<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 4. </p>
</strong>
<div class="figure-caption-contents"><p> Immunostaining (red) of Cx47 in… </p></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"><p> Immunostaining (red) of Cx47 in HeLa-Cx47 ( <i> a </i> ), HeLa wild-type ( <i> b… </i> </p></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">Immunostaining (red) of Cx47 in HeLa-Cx47 (<i>a</i>), HeLa wild-type (<i>b</i>), and HeLa-Cx47-EGFP (<i>c</i>) cells. The EGFP fluorescent signals (<i>d</i>, green) corresponding to Cx47-EGFP fusion proteins were primarily colocalized with Cx47 immunofluorescent signals (<i>c</i>). Typical punctate gap junction staining was detected on contact membranes of Cx47-transfected cells (arrows), whereas no such staining could be detected in HeLa wild-type controls. Nuclei in HeLa-Cx47 and HeLa wild-type cells were stained with TOTO-1 (blue, false color); nuclei in HeLa-Cx47-EGFP cells were not stained but are visible because of higher background staining. Scale bars, 20 μm.</div>
</figcaption>
</figure>
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href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/e4bf84bb9c93/ns1137669005.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="707"
data-image-height="960"
data-image-alt="Figure 5."
data-pmc-id="PMC6740816"
data-figure-id="FIG5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/457a0f490b0a/ns1137669005.gif"
alt="Figure 5." />
</a>
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<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"><p> EGFP signal (green) as reporter… </p></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"><p> EGFP signal (green) as reporter for Cx47-expressing cells and immunostaining (red) for MBP… </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.
</strong>
<div class="figure-caption-contents">EGFP signal (green) as reporter for Cx47-expressing cells and immunostaining (red) for MBP in wild-type (+<i>/</i>+) and homozygous Cx47 <sup>EGFP</sup> (-<i>/</i>-) mice. Shown are double (red, green) confocal images of cerebellar white matter (wm; <i>a, b</i>), corpus callosum (cc; <i>c, d</i>), the funiculus dorsalis (fd) in a transverse section of the spinal cord (<i>e</i>), a longitudinal section of the optic nerve (<i>f</i>), and a longitudinal section of the sciatic nerve (<i>g</i>). All corresponding micrographs (<i>ag</i>) were recorded with the same microscope settings. No EGFP signal was observed in wild-type tissues (<i>a, c</i>) and Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> sciatic nerve (<i>g</i>). Scale bars: <i>a, b, eg</i>, 50 μm; <i>c, d</i>, 100 μm.</div>
</figcaption>
</figure>
<figure class="figure-item "
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data-label-slug="figure-6">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/25920e60d6ec/ns1137669006.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="579"
data-image-height="580"
data-image-alt="Figure 6."
data-pmc-id="PMC6740816"
data-figure-id="FIG6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/01a1064a437d/ns1137669006.gif"
alt="Figure 6." />
</a>
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<strong class="figure-label">
<p> Figure 6. </p>
</strong>
<div class="figure-caption-contents"><p> EGFP signal (green) as reporter… </p></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"><p> EGFP signal (green) as reporter for Cx47-expressing cells and immunostaining (red) for oligodendrocytic… </p></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">EGFP signal (green) as reporter for Cx47-expressing cells and immunostaining (red) for oligodendrocytic CNPase (<i>a</i>), NeuN (<i>b</i>), GFAP (<i>c</i>), and S100β in longitudinal sections of the spinal cord of Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mice. Double (red, green) confocal images are shown. All corresponding images (<i>ad</i>) were recorded with the same microscope settings. CNPase colocalized (yellow) with EGFP fluorescence in the pericaryon (<i>a</i>). Cells immunolabeled for NeuN (<i>b</i>) were EGFP-negative. Likewise, GFAP-positive cells (<i>c</i>) did not express EGFP. A few S100β-positive cells (<i>d</i>) expressed EGFP (yellow); others did not (red), but most EGFP-expressing cells were negative for S100β (green). Scale bars, 50 μm. fl, Funiculus lateralis; gm, gray matter.</div>
</figcaption>
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itemprop="contentUrl"
aria-describedby="figure-caption-6"
role="button"
data-image-width="622"
data-image-height="900"
data-image-alt="Figure 7."
data-pmc-id="PMC6740816"
data-figure-id="FIG7">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-6"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/560a/6740816/ddc8a6197740/ns1137669007.gif"
alt="Figure 7." />
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<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 7. </p>
</strong>
<div class="figure-caption-contents"><p> Electrophysiological properties of GFP-labeled cells… </p></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"><p> Electrophysiological properties of GFP-labeled cells in Cx47 <sup> EGFP( </sup> <sup> + </sup> <sup> / </sup> <sup> - </sup> <sup> ) </sup> and… </p></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">Electrophysiological properties of GFP-labeled cells in Cx47 <sup>EGFP(</sup><sup>+</sup><sup>/</sup><sup>-</sup><sup>)</sup> and PLP-GFP mice. Membrane currents were activated between -160 and +20 mV (50 msec; 10 mV increment; holding potential, -70 mV). <i>a, b</i>, Right, <i>IV</i> plots when taking amplitudes 2 msec (triangles) and 50 msec (diamonds) after the onset of the voltage steps. <i>a</i>, In a fluorescent cell, presumed an oligodendrocyte of the corpus callosum of a Cx47 <sup>EGFP(</sup><sup>+</sup><sup>/</sup><sup>-</sup><sup>)</sup> mouse (resting potential, -60 mV), currents reversed at -67 and -59 mV, respectively. <i>b</i>, A GFP-positive oligodendrocyte in the corpus callosum of a PLP-GFP mouse (resting potential, -70 mV) displayed reversal potentials of -67 and -64 mV. In both cells, currents decayed with similar time constants as indicated (V, +10 mV; a single exponential was fit to the data recorded between 3 and 25 msec after onset of the voltage steps ; time window marked by dashed vertical bars). <i>c</i>, Subsequent to functional characterization, the cytoplasm of the cells shown in <i>a</i> and <i>b</i> was harvested for transcript analysis. PCR products specific to Cx47 (334 bp) were identified by gel electrophoresis (lanes #1, #2 correspond to the cells in <i>a, b</i>, respectively). Cx47 cDNA was completely digested by the restriction endonuclease <i>Mbo</i>I, yielding fragments of 239 and 95 bp (lanes #1c, #2c). <i>d</i>, A GFP-negative CA1 pyramidal neuron (lane #3; PLP-GFP mouse) and an EGFP-negative astrocyte (lane #4; CA1 stratum oriens, Cx47 <sup>EGFP(</sup><sup>+</sup><sup>/</sup><sup>-</sup><sup>)</sup> mouse) were analyzed as described above. Both cells lacked Cx47 transcripts, although mRNA of the housekeeping gene β-actin (238 bp) was found (right). As a length marker, <i>Hinc</i>II-digested DNA from φX174 phages was used.</div>
</figcaption>
</figure>
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id="article-image-7"
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<strong class="figure-label">
<p> Figure 8. </p>
</strong>
<div class="figure-caption-contents"><p> Longitudinal semithin sections of the… </p></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"><p> Longitudinal semithin sections of the optic nerve, close to the eye. The partly… </p></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">Longitudinal semithin sections of the optic nerve, close to the eye. The partly myelinated transition zone of the optic nerve starts 250 μm behind the eye bulb (dashed line). In contrast to the wild-type tissue (<i>a</i>), the myelinated region of the optic nerve from Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mice showed many vacuolated nerve fibers (<i>b</i>). Confocal microscopic analysis of EGFP fluorescence (light gray) in the same region of Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mice (<i>c</i>) demonstrated that Cx47 expression did not occur in front of this transition zone. Scale bars, 100 μm.</div>
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id="article-image-8"
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
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<strong class="figure-label">
<p> Figure 9. </p>
</strong>
<div class="figure-caption-contents"><p> Transverse sections through the partly… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 9. </p>
</strong>
<div class="figure-caption-contents"><p> Transverse sections through the partly myelinated transition zone of the optic nerve. In… </p></div>
</div>
<figcaption id="figure-caption-8" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 9.
</strong>
<div class="figure-caption-contents">Transverse sections through the partly myelinated transition zone of the optic nerve. In wild-type tissue (<i>a</i>) as well as in Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> tissue (<i>b</i>), thin myelin sheets of the first internodes and nonmyelinated nerve fibers were seen. In this transition zone, the vacuolated nerve fibers of the Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> animal were very prominent (asterisks). <i>A</i>, Astrocytes. Scale bars, 3.6 μm.</div>
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data-label-slug="figure-10">
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data-image-width="684"
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-9"
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<strong class="figure-label">
<p> Figure 10. </p>
</strong>
<div class="figure-caption-contents"><p> Transverse sections through the partly… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 10. </p>
</strong>
<div class="figure-caption-contents"><p> Transverse sections through the partly myelinated transition zone of the optic nerve of… </p></div>
</div>
<figcaption id="figure-caption-9" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 10.
</strong>
<div class="figure-caption-contents">Transverse sections through the partly myelinated transition zone of the optic nerve of Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mice as shown in Figure 9. Two types of vacuolated nerve fibers are shown. <i>a</i>, The two nerve fibers exhibit vacuoles (asterisks) between the inner lamellae of the oligodendrocyte (arrows) and the compact lamellae of the myelin sheet. <i>b</i>, The vacuole (asterisk) of this nerve fiber is located between compact myelin sheets. Scale bars, 1.2 μm.</div>
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<strong class="figure-label">
<p> Figure 11. </p>
</strong>
<div class="figure-caption-contents"><p> Transverse semithin sections through the… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 11. </p>
</strong>
<div class="figure-caption-contents"><p> Transverse semithin sections through the intracranial part of the optic nerve immediately in… </p></div>
</div>
<figcaption id="figure-caption-10" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 11.
</strong>
<div class="figure-caption-contents">Transverse semithin sections through the intracranial part of the optic nerve immediately in front of the optic chiasm of 4-week-old Cx32 <sup>(y/</sup><sup>+</sup><sup>)</sup>/Cx47 <sup>(</sup><sup>+</sup><sup>/</sup><sup>+</sup><sup>)</sup> wild-type (wt; <i>a</i>), 5-week-old Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> single-deficient (<i>b</i>), and a Cx32 <sup>(y/</sup><sup>-</sup><sup>)</sup>/Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> double-deficient (<i>c</i>) mice (<i>b, c</i> are littermates). The double-deficient mice exhibited strongly enhanced vacuolation of nerve fibers all over the CNS, whereas the Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mice showed very rare vacuolation (asterisks). Cx47 <sup>EGFP(</sup><sup>-</sup><sup>/</sup><sup>-</sup><sup>)</sup> mice exhibited increased vacuolation only in the transition zone of the optic nerve (Fig. 9<i>b</i>). KO, Knock-out. Scale bars, 16 μm.</div>
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Pathologic and phenotypic alterations in a mouse expressing a connexin47 missense mutation that causes Pelizaeus-Merzbacher-like disease in humans.
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Altevogt BM, Kleopa KA, Postma FR, Scherer SS, Paul DL ( 2002) Connexin29 is uniquely distributed within myelinating glial cells of the central and peripheral nervous systems. J Neurosci 22: 64586470.
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