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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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 |
doi_str_mv | 10.1001/jamasurg.2013.3572 |
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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 < .05), 599 (99) for 8 rats (P < .01), 12.5 (5.5) for 8 rats (P < .01), and 14.0 (3.9) for 8 rats (P < .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 < .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 < .05), 599 (99) for 8 rats (P < .01), 12.5 (5.5) for 8 rats (P < .01), and 14.0 (3.9) for 8 rats (P < .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 < .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 < .05), 599 (99) for 8 rats (P < .01), 12.5 (5.5) for 8 rats (P < .01), and 14.0 (3.9) for 8 rats (P < .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 < .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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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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