The Correlation Between the Expression of Differentiation Markers in Rat Small Intestinal Mucosa and the Transcript Levels of Schlafen 3

IMPORTANCE The normal absorptive function and structural maintenance of the intestinal mucosa depend on a constant process of proliferation of enterocytic stem cells followed by progressive differentiation toward a mature phenotype. The mechanisms that govern enterocytic differentiation in the mucos...

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Veröffentlicht in:JAMA surgery 2013-11, Vol.148 (11), p.1013-1019
Hauptverfasser: Kovalenko, Pavlo L, Basson, Marc D
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Basson, Marc D
description IMPORTANCE The normal absorptive function and structural maintenance of the intestinal mucosa depend on a constant process of proliferation of enterocytic stem cells followed by progressive differentiation toward a mature phenotype. The mechanisms that govern enterocytic differentiation in the mucosa of the small intestine are poorly understood. OBJECTIVE To determine whether schlafen 3 (but not other schlafen proteins) act in vivo and whether its effects are limited to the small intestine. We have previously demonstrated in nonmalignant rat intestinal IEC-6 cells that schlafen 3 levels correlate with the expression of various differentiation markers in vitro in response to differentiation stimuli. DESIGN Randomized controlled experiment. SETTING Animal science laboratory. PARTICIPANTS Male Sprague-Dawley rats 8 to 13 weeks old. MAIN OUTCOMES AND MEASURES Messenger RNA (mRNA) from jejunal and colonic mucosa was isolated, and transcript levels of schlafen proteins 1, 2, 3, 4, 5, 13, and 14; sucrase isomaltase (SI); dipeptidyl peptidase 4 (Dpp4); glucose transporter type 2 (Glut2); and villin were measured by quantitative reverse transcriptase–polymerase chain reaction. We tested parallel variations in protein levels by Western blotting and Dpp4 enzyme activity. RESULTS The transcript level of schlafen 3 (Slfn3) correlated with the levels of the differentiation markers SI, Dpp4, Glut2, and villin. However, the expression of schlafen proteins 1, 2, 4, 5, 13, and 14 did not correlate with the expression of the differentiation markers. The mucosal mRNA levels of Slfn3, SI, Glut2, and Dpp4 were all substantially higher in the rat jejunum than in colonic mucosa by a mean (SE) factor of 51.0 (13.2) for 6 rats (P 
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The mechanisms that govern enterocytic differentiation in the mucosa of the small intestine are poorly understood. OBJECTIVE To determine whether schlafen 3 (but not other schlafen proteins) act in vivo and whether its effects are limited to the small intestine. We have previously demonstrated in nonmalignant rat intestinal IEC-6 cells that schlafen 3 levels correlate with the expression of various differentiation markers in vitro in response to differentiation stimuli. DESIGN Randomized controlled experiment. SETTING Animal science laboratory. PARTICIPANTS Male Sprague-Dawley rats 8 to 13 weeks old. MAIN OUTCOMES AND MEASURES Messenger RNA (mRNA) from jejunal and colonic mucosa was isolated, and transcript levels of schlafen proteins 1, 2, 3, 4, 5, 13, and 14; sucrase isomaltase (SI); dipeptidyl peptidase 4 (Dpp4); glucose transporter type 2 (Glut2); and villin were measured by quantitative reverse transcriptase–polymerase chain reaction. We tested parallel variations in protein levels by Western blotting and Dpp4 enzyme activity. RESULTS The transcript level of schlafen 3 (Slfn3) correlated with the levels of the differentiation markers SI, Dpp4, Glut2, and villin. However, the expression of schlafen proteins 1, 2, 4, 5, 13, and 14 did not correlate with the expression of the differentiation markers. The mucosal mRNA levels of Slfn3, SI, Glut2, and Dpp4 were all substantially higher in the rat jejunum than in colonic mucosa by a mean (SE) factor of 51.0 (13.2) for 6 rats (P &lt; .05), 599 (99) for 8 rats (P &lt; .01), 12.5 (5.5) for 8 rats (P &lt; .01), and 14.0 (3.9) for 8 rats (P &lt; .01), respectively. In IEC-6 cells, infection with adenovirus-expressing GFP-tagged Slfn3 significantly increased Slfn3 expression and Dpp4-specific activity compared with GFP-expressing virus (in 6 rats; P &lt; .05). CONCLUSIONS AND RELEVANCE Taken together with our previous in vitro observations, the results suggest that small intestinal enterocytic epithelial differentiation in rats may be regulated by Slfn3 in vivo, as in vitro, and that these effects may be specific to the small intestinal enterocytic phenotype as opposed to that of the mature colonocyte. Slfn3 human orthologs may be targeted to stimulate intestinal differentiation in patients with short bowel syndrome.</description><identifier>ISSN: 2168-6254</identifier><identifier>EISSN: 2168-6262</identifier><identifier>DOI: 10.1001/jamasurg.2013.3572</identifier><identifier>PMID: 24005468</identifier><language>eng</language><publisher>United States: American Medical Association</publisher><subject>Animals ; Biomarkers - metabolism ; Cell Differentiation - physiology ; Colon - cytology ; Colon - metabolism ; Dipeptidyl Peptidase 4 - genetics ; Dipeptidyl Peptidase 4 - metabolism ; Enterocytes - cytology ; Enterocytes - metabolism ; Glucose Transporter Type 2 - genetics ; Glucose Transporter Type 2 - metabolism ; Ileum - cytology ; Ileum - metabolism ; Jejunum - cytology ; Jejunum - metabolism ; Male ; Microfilament Proteins - genetics ; Microfilament Proteins - metabolism ; Proteins - genetics ; Proteins - metabolism ; Rats ; Rats, Sprague-Dawley ; RNA, Messenger - metabolism ; Sucrase-Isomaltase Complex - genetics ; Sucrase-Isomaltase Complex - metabolism</subject><ispartof>JAMA surgery, 2013-11, Vol.148 (11), p.1013-1019</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a487t-bb310e106c1cb6879c94cce88af9975962301ababf400ea4536caca16e56b8753</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://jamanetwork.com/journals/jamasurgery/articlepdf/10.1001/jamasurg.2013.3572$$EPDF$$P50$$Gama$$H</linktopdf><linktohtml>$$Uhttps://jamanetwork.com/journals/jamasurgery/fullarticle/10.1001/jamasurg.2013.3572$$EHTML$$P50$$Gama$$H</linktohtml><link.rule.ids>64,230,314,780,784,885,3340,27924,27925,76361,76364</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24005468$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kovalenko, Pavlo L</creatorcontrib><creatorcontrib>Basson, Marc D</creatorcontrib><title>The Correlation Between the Expression of Differentiation Markers in Rat Small Intestinal Mucosa and the Transcript Levels of Schlafen 3</title><title>JAMA surgery</title><addtitle>JAMA Surg</addtitle><description>IMPORTANCE The normal absorptive function and structural maintenance of the intestinal mucosa depend on a constant process of proliferation of enterocytic stem cells followed by progressive differentiation toward a mature phenotype. The mechanisms that govern enterocytic differentiation in the mucosa of the small intestine are poorly understood. OBJECTIVE To determine whether schlafen 3 (but not other schlafen proteins) act in vivo and whether its effects are limited to the small intestine. We have previously demonstrated in nonmalignant rat intestinal IEC-6 cells that schlafen 3 levels correlate with the expression of various differentiation markers in vitro in response to differentiation stimuli. DESIGN Randomized controlled experiment. SETTING Animal science laboratory. PARTICIPANTS Male Sprague-Dawley rats 8 to 13 weeks old. MAIN OUTCOMES AND MEASURES Messenger RNA (mRNA) from jejunal and colonic mucosa was isolated, and transcript levels of schlafen proteins 1, 2, 3, 4, 5, 13, and 14; sucrase isomaltase (SI); dipeptidyl peptidase 4 (Dpp4); glucose transporter type 2 (Glut2); and villin were measured by quantitative reverse transcriptase–polymerase chain reaction. We tested parallel variations in protein levels by Western blotting and Dpp4 enzyme activity. RESULTS The transcript level of schlafen 3 (Slfn3) correlated with the levels of the differentiation markers SI, Dpp4, Glut2, and villin. However, the expression of schlafen proteins 1, 2, 4, 5, 13, and 14 did not correlate with the expression of the differentiation markers. The mucosal mRNA levels of Slfn3, SI, Glut2, and Dpp4 were all substantially higher in the rat jejunum than in colonic mucosa by a mean (SE) factor of 51.0 (13.2) for 6 rats (P &lt; .05), 599 (99) for 8 rats (P &lt; .01), 12.5 (5.5) for 8 rats (P &lt; .01), and 14.0 (3.9) for 8 rats (P &lt; .01), respectively. In IEC-6 cells, infection with adenovirus-expressing GFP-tagged Slfn3 significantly increased Slfn3 expression and Dpp4-specific activity compared with GFP-expressing virus (in 6 rats; P &lt; .05). CONCLUSIONS AND RELEVANCE Taken together with our previous in vitro observations, the results suggest that small intestinal enterocytic epithelial differentiation in rats may be regulated by Slfn3 in vivo, as in vitro, and that these effects may be specific to the small intestinal enterocytic phenotype as opposed to that of the mature colonocyte. Slfn3 human orthologs may be targeted to stimulate intestinal differentiation in patients with short bowel syndrome.</description><subject>Animals</subject><subject>Biomarkers - metabolism</subject><subject>Cell Differentiation - physiology</subject><subject>Colon - cytology</subject><subject>Colon - metabolism</subject><subject>Dipeptidyl Peptidase 4 - genetics</subject><subject>Dipeptidyl Peptidase 4 - metabolism</subject><subject>Enterocytes - cytology</subject><subject>Enterocytes - metabolism</subject><subject>Glucose Transporter Type 2 - genetics</subject><subject>Glucose Transporter Type 2 - metabolism</subject><subject>Ileum - cytology</subject><subject>Ileum - metabolism</subject><subject>Jejunum - cytology</subject><subject>Jejunum - metabolism</subject><subject>Male</subject><subject>Microfilament Proteins - genetics</subject><subject>Microfilament Proteins - metabolism</subject><subject>Proteins - genetics</subject><subject>Proteins - metabolism</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>RNA, Messenger - metabolism</subject><subject>Sucrase-Isomaltase Complex - genetics</subject><subject>Sucrase-Isomaltase Complex - metabolism</subject><issn>2168-6254</issn><issn>2168-6262</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkc9u1DAQxiMEolXpC_SAfOSyi__FcS5IsC1QaSskupytiTvpujjOYjsF3oDHxmHbBXyxNf7mN_Ppq6ozRpeMUvb6DgZIU7xdcsrEUtQNf1Idc6b0QnHFnx7etTyqTlO6o-VoSqVon1dHXFJaS6WPq1-bLZLVGCN6yG4M5B3m74iB5FK_-LGLmNJcHnty7voeI4bs9soriF8xJuIC-QyZXA_gPbkMGVN2ATy5muyYgEC4-QPbRAjJRrfLZI336NPMvLZbD30ZJ15Uz3rwCU8f7pPqy_uLzerjYv3pw-Xq7XoBUjd50XWCUWRUWWY7pZvWttJa1Br6tm3qVnFBGXTQ9cUigqyFsmCBKaxVp5tanFRv9tzd1A14Y4ufCN7sohsg_jQjOPP_T3BbczveG1m3tNUz4NUDII7fpmLWDC5Z9B4CjlMyTComJK81K1K-l9o4phSxP4xh1MwpmscUzZyimVMsTS__XfDQ8phZEZztBaX3L7ARUrZa_Aa-SaW3</recordid><startdate>20131101</startdate><enddate>20131101</enddate><creator>Kovalenko, Pavlo L</creator><creator>Basson, Marc D</creator><general>American Medical Association</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20131101</creationdate><title>The Correlation Between the Expression of Differentiation Markers in Rat Small Intestinal Mucosa and the Transcript Levels of Schlafen 3</title><author>Kovalenko, Pavlo L ; Basson, Marc D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a487t-bb310e106c1cb6879c94cce88af9975962301ababf400ea4536caca16e56b8753</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Biomarkers - metabolism</topic><topic>Cell Differentiation - physiology</topic><topic>Colon - cytology</topic><topic>Colon - metabolism</topic><topic>Dipeptidyl Peptidase 4 - genetics</topic><topic>Dipeptidyl Peptidase 4 - metabolism</topic><topic>Enterocytes - cytology</topic><topic>Enterocytes - metabolism</topic><topic>Glucose Transporter Type 2 - genetics</topic><topic>Glucose Transporter Type 2 - metabolism</topic><topic>Ileum - cytology</topic><topic>Ileum - metabolism</topic><topic>Jejunum - cytology</topic><topic>Jejunum - metabolism</topic><topic>Male</topic><topic>Microfilament Proteins - genetics</topic><topic>Microfilament Proteins - metabolism</topic><topic>Proteins - genetics</topic><topic>Proteins - metabolism</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>RNA, Messenger - metabolism</topic><topic>Sucrase-Isomaltase Complex - genetics</topic><topic>Sucrase-Isomaltase Complex - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kovalenko, Pavlo L</creatorcontrib><creatorcontrib>Basson, Marc D</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>JAMA surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kovalenko, Pavlo L</au><au>Basson, Marc D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Correlation Between the Expression of Differentiation Markers in Rat Small Intestinal Mucosa and the Transcript Levels of Schlafen 3</atitle><jtitle>JAMA surgery</jtitle><addtitle>JAMA Surg</addtitle><date>2013-11-01</date><risdate>2013</risdate><volume>148</volume><issue>11</issue><spage>1013</spage><epage>1019</epage><pages>1013-1019</pages><issn>2168-6254</issn><eissn>2168-6262</eissn><abstract>IMPORTANCE The normal absorptive function and structural maintenance of the intestinal mucosa depend on a constant process of proliferation of enterocytic stem cells followed by progressive differentiation toward a mature phenotype. The mechanisms that govern enterocytic differentiation in the mucosa of the small intestine are poorly understood. OBJECTIVE To determine whether schlafen 3 (but not other schlafen proteins) act in vivo and whether its effects are limited to the small intestine. We have previously demonstrated in nonmalignant rat intestinal IEC-6 cells that schlafen 3 levels correlate with the expression of various differentiation markers in vitro in response to differentiation stimuli. DESIGN Randomized controlled experiment. SETTING Animal science laboratory. PARTICIPANTS Male Sprague-Dawley rats 8 to 13 weeks old. MAIN OUTCOMES AND MEASURES Messenger RNA (mRNA) from jejunal and colonic mucosa was isolated, and transcript levels of schlafen proteins 1, 2, 3, 4, 5, 13, and 14; sucrase isomaltase (SI); dipeptidyl peptidase 4 (Dpp4); glucose transporter type 2 (Glut2); and villin were measured by quantitative reverse transcriptase–polymerase chain reaction. We tested parallel variations in protein levels by Western blotting and Dpp4 enzyme activity. RESULTS The transcript level of schlafen 3 (Slfn3) correlated with the levels of the differentiation markers SI, Dpp4, Glut2, and villin. However, the expression of schlafen proteins 1, 2, 4, 5, 13, and 14 did not correlate with the expression of the differentiation markers. The mucosal mRNA levels of Slfn3, SI, Glut2, and Dpp4 were all substantially higher in the rat jejunum than in colonic mucosa by a mean (SE) factor of 51.0 (13.2) for 6 rats (P &lt; .05), 599 (99) for 8 rats (P &lt; .01), 12.5 (5.5) for 8 rats (P &lt; .01), and 14.0 (3.9) for 8 rats (P &lt; .01), respectively. In IEC-6 cells, infection with adenovirus-expressing GFP-tagged Slfn3 significantly increased Slfn3 expression and Dpp4-specific activity compared with GFP-expressing virus (in 6 rats; P &lt; .05). CONCLUSIONS AND RELEVANCE Taken together with our previous in vitro observations, the results suggest that small intestinal enterocytic epithelial differentiation in rats may be regulated by Slfn3 in vivo, as in vitro, and that these effects may be specific to the small intestinal enterocytic phenotype as opposed to that of the mature colonocyte. Slfn3 human orthologs may be targeted to stimulate intestinal differentiation in patients with short bowel syndrome.</abstract><cop>United States</cop><pub>American Medical Association</pub><pmid>24005468</pmid><doi>10.1001/jamasurg.2013.3572</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record>
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source MEDLINE; American Medical Association Journal
subjects Animals
Biomarkers - metabolism
Cell Differentiation - physiology
Colon - cytology
Colon - metabolism
Dipeptidyl Peptidase 4 - genetics
Dipeptidyl Peptidase 4 - metabolism
Enterocytes - cytology
Enterocytes - metabolism
Glucose Transporter Type 2 - genetics
Glucose Transporter Type 2 - metabolism
Ileum - cytology
Ileum - metabolism
Jejunum - cytology
Jejunum - metabolism
Male
Microfilament Proteins - genetics
Microfilament Proteins - metabolism
Proteins - genetics
Proteins - metabolism
Rats
Rats, Sprague-Dawley
RNA, Messenger - metabolism
Sucrase-Isomaltase Complex - genetics
Sucrase-Isomaltase Complex - metabolism
title The Correlation Between the Expression of Differentiation Markers in Rat Small Intestinal Mucosa and the Transcript Levels of Schlafen 3
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