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Serial block-face scanning electron microscopy reveals neuronal-epithelial cell fusion in the mouse cornea

Identifieur interne : 000392 ( Pmc/Checkpoint ); précédent : 000391; suivant : 000393

Serial block-face scanning electron microscopy reveals neuronal-epithelial cell fusion in the mouse cornea

Auteurs : Justin A. Courson [États-Unis] ; Ian Smith [États-Unis] ; Thao Do [États-Unis] ; Paul T. Landry [États-Unis] ; Aubrey Hargrave [États-Unis] ; Ali R. Behzad [Arabie saoudite] ; Sam D. Hanlon [États-Unis] ; Rolando E. Rumbaut [États-Unis] ; C. Wayne Smith [États-Unis] ; Alan R. Burns [États-Unis]

Source :

RBID : PMC:6853292

Abstract

The cornea is the most highly innervated tissue in the body. It is generally accepted that corneal stromal nerves penetrate the epithelial basal lamina giving rise to intra-epithelial nerves. During the course of a study wherein we imaged corneal nerves in mice, we observed a novel neuronal-epithelial cell interaction whereby nerves approaching the epithelium in the cornea fused with basal epithelial cells, such that their plasma membranes were continuous and the neuronal axoplasm freely abutted the epithelial cytoplasm. In this study we sought to determine the frequency, distribution, and morphological profile of neuronal-epithelial cell fusion events within the cornea. Serial electron microscopy images were obtained from the anterior stroma in the paralimbus and central cornea of 8–10 week old C57BL/6J mice. We found evidence of a novel alternative behavior involving a neuronal-epithelial interaction whereby 42.8% of central corneal nerve bundles approaching the epithelium contain axons that fuse with basal epithelial cells. The average surface-to-volume ratio of a penetrating nerve was 3.32, while the average fusing nerve was smaller at 1.39 (p ≤ 0.0001). Despite this, both neuronal-epithelial cell interactions involve similarly sized discontinuities in the basal lamina. In order to verify the plasma membrane continuity between fused neurons and epithelial cells we used the lipophilic membrane tracer DiI. The majority of corneal nerves were labeled with DiI after application to the trigeminal ganglion and, consistent with our ultrastructural observations, fusion sites recognized as DiI-labeled basal epithelial cells were located at points of stromal nerve termination. These studies provide evidence that neuronal-epithelial cell fusion is a cell-cell interaction that occurs primarily in the central cornea, and fusing nerve bundles are morphologically distinct from penetrating nerve bundles. This is, to our knowledge, the first description of neuronal-epithelial cell fusion in the literature adding a new level of complexity to the current understanding of corneal innervation.


Url:
DOI: 10.1371/journal.pone.0224434
PubMed: 31721785
PubMed Central: 6853292


Affiliations:


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PMC:6853292

Le document en format XML

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<p>The cornea is the most highly innervated tissue in the body. It is generally accepted that corneal stromal nerves penetrate the epithelial basal lamina giving rise to intra-epithelial nerves. During the course of a study wherein we imaged corneal nerves in mice, we observed a novel neuronal-epithelial cell interaction whereby nerves approaching the epithelium in the cornea fused with basal epithelial cells, such that their plasma membranes were continuous and the neuronal axoplasm freely abutted the epithelial cytoplasm. In this study we sought to determine the frequency, distribution, and morphological profile of neuronal-epithelial cell fusion events within the cornea. Serial electron microscopy images were obtained from the anterior stroma in the paralimbus and central cornea of 8–10 week old C57BL/6J mice. We found evidence of a novel alternative behavior involving a neuronal-epithelial interaction whereby 42.8% of central corneal nerve bundles approaching the epithelium contain axons that fuse with basal epithelial cells. The average surface-to-volume ratio of a penetrating nerve was 3.32, while the average fusing nerve was smaller at 1.39 (p ≤ 0.0001). Despite this, both neuronal-epithelial cell interactions involve similarly sized discontinuities in the basal lamina. In order to verify the plasma membrane continuity between fused neurons and epithelial cells we used the lipophilic membrane tracer DiI. The majority of corneal nerves were labeled with DiI after application to the trigeminal ganglion and, consistent with our ultrastructural observations, fusion sites recognized as DiI-labeled basal epithelial cells were located at points of stromal nerve termination. These studies provide evidence that neuronal-epithelial cell fusion is a cell-cell interaction that occurs primarily in the central cornea, and fusing nerve bundles are morphologically distinct from penetrating nerve bundles. This is, to our knowledge, the first description of neuronal-epithelial cell fusion in the literature adding a new level of complexity to the current understanding of corneal innervation.</p>
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<name>
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<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="corresp" rid="cor001">*</xref>
</contrib>
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<name>
<surname>Smith</surname>
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</name>
<role content-type="http://credit.casrai.org/">Visualization</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
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<name>
<surname>Do</surname>
<given-names>Thao</given-names>
</name>
<role content-type="http://credit.casrai.org/">Visualization</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
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<name>
<surname>Landry</surname>
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</name>
<role content-type="http://credit.casrai.org/">Methodology</role>
<role content-type="http://credit.casrai.org/">Visualization</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
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<name>
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<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
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<name>
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<role content-type="http://credit.casrai.org/">Validation</role>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
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<contrib contrib-type="author">
<name>
<surname>Hanlon</surname>
<given-names>Sam D.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Methodology</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Rumbaut</surname>
<given-names>Rolando E.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Funding acquisition</role>
<role content-type="http://credit.casrai.org/">Resources</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
<xref ref-type="aff" rid="aff004">
<sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Smith</surname>
<given-names>C. Wayne</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Funding acquisition</role>
<role content-type="http://credit.casrai.org/">Resources</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Burns</surname>
<given-names>Alan R.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Funding acquisition</role>
<role content-type="http://credit.casrai.org/">Project administration</role>
<role content-type="http://credit.casrai.org/">Resources</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
</contrib>
</contrib-group>
<aff id="aff001">
<label>1</label>
<addr-line>University of Houston, College of Optometry, Houston, TX, United States of America</addr-line>
</aff>
<aff id="aff002">
<label>2</label>
<addr-line>King Abdullah University of Science and Technology (KAUST), Core Labs, Thuwal, Saudi Arabia</addr-line>
</aff>
<aff id="aff003">
<label>3</label>
<addr-line>Baylor College of Medicine, Children’s Nutrition Center, Houston, TX, United States of America</addr-line>
</aff>
<aff id="aff004">
<label>4</label>
<addr-line>Center for Translational Research on Inflammatory Diseases (CTRID), Michael E. DeBakey Veterans Affairs Medical Center, Houston, TX, United States of America</addr-line>
</aff>
<contrib-group>
<contrib contrib-type="editor">
<name>
<surname>Lewin</surname>
<given-names>Alfred S.</given-names>
</name>
<role>Editor</role>
<xref ref-type="aff" rid="edit1"></xref>
</contrib>
</contrib-group>
<aff id="edit1">
<addr-line>University of Florida, UNITED STATES</addr-line>
</aff>
<author-notes>
<fn fn-type="COI-statement" id="coi001">
<p>
<bold>Competing Interests: </bold>
The authors have declared that no competing interests exist.</p>
</fn>
<corresp id="cor001">* E-mail:
<email>JACourso@central.uh.edu</email>
</corresp>
</author-notes>
<pub-date pub-type="epub">
<day>13</day>
<month>11</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="collection">
<year>2019</year>
</pub-date>
<volume>14</volume>
<issue>11</issue>
<elocation-id>e0224434</elocation-id>
<history>
<date date-type="received">
<day>16</day>
<month>8</month>
<year>2019</year>
</date>
<date date-type="accepted">
<day>14</day>
<month>10</month>
<year>2019</year>
</date>
</history>
<permissions>
<copyright-statement>© 2019 Courson et al</copyright-statement>
<copyright-year>2019</copyright-year>
<copyright-holder>Courson et al</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<license-p>This is an open access article distributed under the terms of the
<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>
, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p>
</license>
</permissions>
<self-uri content-type="pdf" xlink:href="pone.0224434.pdf"></self-uri>
<abstract>
<p>The cornea is the most highly innervated tissue in the body. It is generally accepted that corneal stromal nerves penetrate the epithelial basal lamina giving rise to intra-epithelial nerves. During the course of a study wherein we imaged corneal nerves in mice, we observed a novel neuronal-epithelial cell interaction whereby nerves approaching the epithelium in the cornea fused with basal epithelial cells, such that their plasma membranes were continuous and the neuronal axoplasm freely abutted the epithelial cytoplasm. In this study we sought to determine the frequency, distribution, and morphological profile of neuronal-epithelial cell fusion events within the cornea. Serial electron microscopy images were obtained from the anterior stroma in the paralimbus and central cornea of 8–10 week old C57BL/6J mice. We found evidence of a novel alternative behavior involving a neuronal-epithelial interaction whereby 42.8% of central corneal nerve bundles approaching the epithelium contain axons that fuse with basal epithelial cells. The average surface-to-volume ratio of a penetrating nerve was 3.32, while the average fusing nerve was smaller at 1.39 (p ≤ 0.0001). Despite this, both neuronal-epithelial cell interactions involve similarly sized discontinuities in the basal lamina. In order to verify the plasma membrane continuity between fused neurons and epithelial cells we used the lipophilic membrane tracer DiI. The majority of corneal nerves were labeled with DiI after application to the trigeminal ganglion and, consistent with our ultrastructural observations, fusion sites recognized as DiI-labeled basal epithelial cells were located at points of stromal nerve termination. These studies provide evidence that neuronal-epithelial cell fusion is a cell-cell interaction that occurs primarily in the central cornea, and fusing nerve bundles are morphologically distinct from penetrating nerve bundles. This is, to our knowledge, the first description of neuronal-epithelial cell fusion in the literature adding a new level of complexity to the current understanding of corneal innervation.</p>
</abstract>
<funding-group>
<funding-statement>Funded by R01 EY018239. Agency: National Institutes of Health (NIH). Institute: National Eye Institute (NEI). Type: Research Project (R01). Funding number: EY018239. P30 EY07551. Agency: National Institutes of Health (NIH). Institute: National Eye Institute (NEI). Type: Center Core Grant. Funding Number: EY07551. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement>
</funding-group>
<counts>
<fig-count count="12"></fig-count>
<table-count count="0"></table-count>
<page-count count="24"></page-count>
</counts>
<custom-meta-group>
<custom-meta id="data-availability">
<meta-name>Data Availability</meta-name>
<meta-value>All relevant data are within the paper and its Supporting Information files.</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
<notes>
<title>Data Availability</title>
<p>All relevant data are within the paper and its Supporting Information files.</p>
</notes>
</front>
</pmc>
<affiliations>
<list>
<country>
<li>Arabie saoudite</li>
<li>États-Unis</li>
</country>
</list>
<tree>
<country name="États-Unis">
<noRegion>
<name sortKey="Courson, Justin A" sort="Courson, Justin A" uniqKey="Courson J" first="Justin A." last="Courson">Justin A. Courson</name>
</noRegion>
<name sortKey="Burns, Alan R" sort="Burns, Alan R" uniqKey="Burns A" first="Alan R." last="Burns">Alan R. Burns</name>
<name sortKey="Burns, Alan R" sort="Burns, Alan R" uniqKey="Burns A" first="Alan R." last="Burns">Alan R. Burns</name>
<name sortKey="Do, Thao" sort="Do, Thao" uniqKey="Do T" first="Thao" last="Do">Thao Do</name>
<name sortKey="Hanlon, Sam D" sort="Hanlon, Sam D" uniqKey="Hanlon S" first="Sam D." last="Hanlon">Sam D. Hanlon</name>
<name sortKey="Hargrave, Aubrey" sort="Hargrave, Aubrey" uniqKey="Hargrave A" first="Aubrey" last="Hargrave">Aubrey Hargrave</name>
<name sortKey="Landry, Paul T" sort="Landry, Paul T" uniqKey="Landry P" first="Paul T." last="Landry">Paul T. Landry</name>
<name sortKey="Rumbaut, Rolando E" sort="Rumbaut, Rolando E" uniqKey="Rumbaut R" first="Rolando E." last="Rumbaut">Rolando E. Rumbaut</name>
<name sortKey="Rumbaut, Rolando E" sort="Rumbaut, Rolando E" uniqKey="Rumbaut R" first="Rolando E." last="Rumbaut">Rolando E. Rumbaut</name>
<name sortKey="Smith, C Wayne" sort="Smith, C Wayne" uniqKey="Smith C" first="C. Wayne" last="Smith">C. Wayne Smith</name>
<name sortKey="Smith, Ian" sort="Smith, Ian" uniqKey="Smith I" first="Ian" last="Smith">Ian Smith</name>
</country>
<country name="Arabie saoudite">
<noRegion>
<name sortKey="Behzad, Ali R" sort="Behzad, Ali R" uniqKey="Behzad A" first="Ali R." last="Behzad">Ali R. Behzad</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>

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