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Regulation of epithelial differentiation in rat intestine by intraluminal delivery of an adenoviral vector or silencing RNA coding for Schlafen 3.

Kovalenko PL, Yuan L, Sun K, Kunovska L, Seregin S, Amalfitano A, Basson MD - PLoS ONE (2013)

Bottom Line: This Slfn3 overexpression was associated with increases in all four differentiation markers.Injecting siSlfn3 into rat jejunum in vivo substantially reduced Slfn3 and all four intestinal mucosal differentiation markers three days later, as well as Dpp4 specific activity.Endogenous Slfn3 was reduced in atrophic mucosa from a blind-end Roux-en-Y anastomosis in parallel with differentiation marker expression together with AKT and p38 signaling.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Michigan State University, East Lansing, Michigan, United States of America.

ABSTRACT
Although we stimulate enterocytic proliferation to ameliorate short gut syndrome or mucosal atrophy, less effort has been directed at enterocytic differentiation. Schlafen 3 (Slfn3) is a poorly understood protein induced during IEC-6 enterocytic differentiation. We hypothesized that exogenous manipulation of Slfn3 would regulate enterocytic differentiation in vivo. Adenoviral vector coding for Slfn3 cDNA (Ad-GFP-Slfn3) or silencing RNA for Slfn3 (siSlfn3) was introduced intraluminally into rat intestine. We assessed Slfn3, villin, sucrase-isomaltase (SI), Dpp4, and Glut2 by qRT-PCR, Western blot, and immunohistochemistry. We also studied Slfn3 and these differentiation markers in atrophic defunctionalized jejunal mucosa and the crypt-villus axis of normal jejunum. Ad-GFP-Slfn3 but not Ad-GFP increased Slfn3, villin and Dpp4 expression in human Caco-2 intestinal epithelial cells. Injecting Ad-GFP-Slfn3 into rat jejunum in vivo increased mucosal Slfn3 mRNA three days later vs. intraluminal Ad-GFP. This Slfn3 overexpression was associated with increases in all four differentiation markers. Injecting siSlfn3 into rat jejunum in vivo substantially reduced Slfn3 and all four intestinal mucosal differentiation markers three days later, as well as Dpp4 specific activity. Endogenous Slfn3 was reduced in atrophic mucosa from a blind-end Roux-en-Y anastomosis in parallel with differentiation marker expression together with AKT and p38 signaling. Slfn3 was more highly expressed in the villi than the crypts, paralleling Glut2, SI and Dpp4. Slfn3 is a key intracellular regulator of rat enterocytic differentiation. Understanding how Slfn3 works may identify targets to promote enterocytic differentiation and maintain mucosal function in vivo, facilitating enteral nutrition and improving survival in patients with mucosal atrophy or short gut syndrome.

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Related in: MedlinePlus

Difference in differentiation markers expression within intestinal mucosa.The level of Slfn3 was significantly lower in crypts of mucosa compared to villi (p<0.05, n = 6). The transcript levels of Dpp4 (B), SI (C), Glut2 (D) but not villin (E) were significantly lower in jejunal crypts compared to villi (p<0.05, n = 6).
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pone-0079745-g006: Difference in differentiation markers expression within intestinal mucosa.The level of Slfn3 was significantly lower in crypts of mucosa compared to villi (p<0.05, n = 6). The transcript levels of Dpp4 (B), SI (C), Glut2 (D) but not villin (E) were significantly lower in jejunal crypts compared to villi (p<0.05, n = 6).

Mentions: In further studies, we used LCM to compare the expression of Slfn3 and of some enterocytic differentiation markers between the intestinal epithelial cells of the intestinal crypts and villi in normal rat jejunum. Slfn3 mRNA was 5.2±0.9 fold higher in the villi of the jejunal mucosa than in the crypts (Fig 6A, p<0.05, n = 6). Similarly expression of Dpp4 (Fig 6B), SI (Fig 6C) and Glut2 (Fig 6D) were 5.4±0.7, 6.7±0.8 and 9.1±0.5 fold higher respectively in the villi compared to the crypts. However no difference between crypts and villi was observed in the transcript level of villin (Fig 6E).


Regulation of epithelial differentiation in rat intestine by intraluminal delivery of an adenoviral vector or silencing RNA coding for Schlafen 3.

Kovalenko PL, Yuan L, Sun K, Kunovska L, Seregin S, Amalfitano A, Basson MD - PLoS ONE (2013)

Difference in differentiation markers expression within intestinal mucosa.The level of Slfn3 was significantly lower in crypts of mucosa compared to villi (p<0.05, n = 6). The transcript levels of Dpp4 (B), SI (C), Glut2 (D) but not villin (E) were significantly lower in jejunal crypts compared to villi (p<0.05, n = 6).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3823574&req=5

pone-0079745-g006: Difference in differentiation markers expression within intestinal mucosa.The level of Slfn3 was significantly lower in crypts of mucosa compared to villi (p<0.05, n = 6). The transcript levels of Dpp4 (B), SI (C), Glut2 (D) but not villin (E) were significantly lower in jejunal crypts compared to villi (p<0.05, n = 6).
Mentions: In further studies, we used LCM to compare the expression of Slfn3 and of some enterocytic differentiation markers between the intestinal epithelial cells of the intestinal crypts and villi in normal rat jejunum. Slfn3 mRNA was 5.2±0.9 fold higher in the villi of the jejunal mucosa than in the crypts (Fig 6A, p<0.05, n = 6). Similarly expression of Dpp4 (Fig 6B), SI (Fig 6C) and Glut2 (Fig 6D) were 5.4±0.7, 6.7±0.8 and 9.1±0.5 fold higher respectively in the villi compared to the crypts. However no difference between crypts and villi was observed in the transcript level of villin (Fig 6E).

Bottom Line: This Slfn3 overexpression was associated with increases in all four differentiation markers.Injecting siSlfn3 into rat jejunum in vivo substantially reduced Slfn3 and all four intestinal mucosal differentiation markers three days later, as well as Dpp4 specific activity.Endogenous Slfn3 was reduced in atrophic mucosa from a blind-end Roux-en-Y anastomosis in parallel with differentiation marker expression together with AKT and p38 signaling.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Michigan State University, East Lansing, Michigan, United States of America.

ABSTRACT
Although we stimulate enterocytic proliferation to ameliorate short gut syndrome or mucosal atrophy, less effort has been directed at enterocytic differentiation. Schlafen 3 (Slfn3) is a poorly understood protein induced during IEC-6 enterocytic differentiation. We hypothesized that exogenous manipulation of Slfn3 would regulate enterocytic differentiation in vivo. Adenoviral vector coding for Slfn3 cDNA (Ad-GFP-Slfn3) or silencing RNA for Slfn3 (siSlfn3) was introduced intraluminally into rat intestine. We assessed Slfn3, villin, sucrase-isomaltase (SI), Dpp4, and Glut2 by qRT-PCR, Western blot, and immunohistochemistry. We also studied Slfn3 and these differentiation markers in atrophic defunctionalized jejunal mucosa and the crypt-villus axis of normal jejunum. Ad-GFP-Slfn3 but not Ad-GFP increased Slfn3, villin and Dpp4 expression in human Caco-2 intestinal epithelial cells. Injecting Ad-GFP-Slfn3 into rat jejunum in vivo increased mucosal Slfn3 mRNA three days later vs. intraluminal Ad-GFP. This Slfn3 overexpression was associated with increases in all four differentiation markers. Injecting siSlfn3 into rat jejunum in vivo substantially reduced Slfn3 and all four intestinal mucosal differentiation markers three days later, as well as Dpp4 specific activity. Endogenous Slfn3 was reduced in atrophic mucosa from a blind-end Roux-en-Y anastomosis in parallel with differentiation marker expression together with AKT and p38 signaling. Slfn3 was more highly expressed in the villi than the crypts, paralleling Glut2, SI and Dpp4. Slfn3 is a key intracellular regulator of rat enterocytic differentiation. Understanding how Slfn3 works may identify targets to promote enterocytic differentiation and maintain mucosal function in vivo, facilitating enteral nutrition and improving survival in patients with mucosal atrophy or short gut syndrome.

Show MeSH
Related in: MedlinePlus