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Alterations in content and localization of defensins in rat ileum and jejunum following ischemia-reperfusion. Specific peptides, in specific places, for specific jobs?

Kozar RA, Santora RJ, Poindexter BJ, Milner SM, Bick RJ - Eplasty (2011)

Bottom Line: Shams were subjected to laparotomy but no clamping.Ileum and jejunum were harvested and sectioned, and subjected to fluorescence deconvolution microscopy for determinations of content and localization of rat beta defensins, 1, 2, 3; rat neutrophil protein-1; and cathelicidin LL-37.Ischemia-reperfusion increased neutrophil defensin alpha (RNP-1) in jejunum; rat beta defensin 1 was increased 2-fold in ileal mucosa and slightly reduced in jejunal mucosa; rat beta defensin 2 was reduced by ischemia-reperfusion in ileum, but slightly increased in jejunum; rat beta defensin 3 was concentrated in the muscularis externa and myenteric plexus of the jejunum; ischemia-reperfusion did not alter cathelicidin LL-37 content in the small intestine, although a greater concentration was seen in jejunum compared with ileum.

View Article: PubMed Central - PubMed

ABSTRACT

Objective: To determine alterations in quantities and distributions of natural antimicrobials following ischemia-reperfusion injury. We hypothesized that these compounds would be upregulated in areas of small intestine where changes in permeability and cellular disruption were likely and where protective mechanisms would be initiated.

Methods: Rats with ischemia-reperfusion underwent superior mesenteric artery clamping and reperfusion. Shams were subjected to laparotomy but no clamping. Ileum and jejunum were harvested and sectioned, and subjected to fluorescence deconvolution microscopy for determinations of content and localization of rat beta defensins, 1, 2, 3; rat neutrophil protein-1; and cathelicidin LL-37. Modeling was performed to determine cellular location of antimicrobials.

Results: Ischemia-reperfusion increased neutrophil defensin alpha (RNP-1) in jejunum; rat beta defensin 1 was increased 2-fold in ileal mucosa and slightly reduced in jejunal mucosa; rat beta defensin 2 was reduced by ischemia-reperfusion in ileum, but slightly increased in jejunum; rat beta defensin 3 was concentrated in the muscularis externa and myenteric plexus of the jejunum; ischemia-reperfusion did not alter cathelicidin LL-37 content in the small intestine, although a greater concentration was seen in jejunum compared with ileum.

Conclusion: Ischemia-reperfusion injury caused changes in antimicrobial content in defined areas, and these different regulations might reflect the specific roles of jejunum versus ileum.

No MeSH data available.


Related in: MedlinePlus

This figure demonstrates the specificities with regard to both concentration changes and localizations of various antimicrobials in the ileum. Each pair of images (panels A through F) shows the sham tissue on the left and the ischemia-reperfusion image on the right. The first 2 panels (A) are images used to produce Figure 1. More rat beta defensin (RBD)-1 is seen in the apical and middle regions of the villi. Panel B again shows RBD-1 (red), but this time in the base/neck region of the villi. Panel C shows RBD-3 before and after ischemia-reperfusion demonstrating a decrease in peptide in the outer, longitudinal muscularis externa and a small increase in the inner, circular layer. Panel D shows RBD-2 increase in enterocytes due to I/R and a decrease in perilacteal concentration. Panel E shows RNP-1 throughout the epithelial layer of both sham and ischemia-reperfusion samples; panel F clearly demonstrates a reduction of cathelicidin in the outer, longitudinal muscularis externa, but an increase in the lower, glandular aspects of the mucosa (magnification X 400).
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Figure 2: This figure demonstrates the specificities with regard to both concentration changes and localizations of various antimicrobials in the ileum. Each pair of images (panels A through F) shows the sham tissue on the left and the ischemia-reperfusion image on the right. The first 2 panels (A) are images used to produce Figure 1. More rat beta defensin (RBD)-1 is seen in the apical and middle regions of the villi. Panel B again shows RBD-1 (red), but this time in the base/neck region of the villi. Panel C shows RBD-3 before and after ischemia-reperfusion demonstrating a decrease in peptide in the outer, longitudinal muscularis externa and a small increase in the inner, circular layer. Panel D shows RBD-2 increase in enterocytes due to I/R and a decrease in perilacteal concentration. Panel E shows RNP-1 throughout the epithelial layer of both sham and ischemia-reperfusion samples; panel F clearly demonstrates a reduction of cathelicidin in the outer, longitudinal muscularis externa, but an increase in the lower, glandular aspects of the mucosa (magnification X 400).

Mentions: To further show the regulations and specificities, Figure 2 includes 6 pairs of images (shams on the left and I/R samples on the right). In each of the panels, the antimicrobial peptide is shown in red (orange/yellow where colocalization occurs). Cell nuclei are blue and F-actin is green. The first pair (panel A) are the images from Figure 1, but of note here is the high concentration of RBD-1 seen in the lower portion of the villi in the I/R sample. Panel B also shows RBD-1, but in the lower portion of the mucosa, where the crypts meet the glands. Note the increased amount of (red) peptide following I/R. Panel C is of RBD-3, located at an extremely high concentration in the longitudinal layer of the muscularis externa, but greatly reduced following I/R. However, this loss is somewhat compensated for by an increase in the inner, circular layer of the externa and maybe even in the myenteric plexus. Panel D demonstrates that I/R injury shows a trend of increasing RBD-2 in luminal epithelial cells in jejunum as seen in this pair of images, but not in ileum. Overall, the values are not significantly different; however, as stated, the trend within pairs is for an increase in staining intensity as shown here (13300 ± 7889 vs 16123 ± 7066; sham vs I/R pixels). Panel E shows that there was no change (18938 ± 8607 vs 18245 ± 7168; sham vs I/R; pixels) when comparing concentrations of RNP-1. The final pair of images, panel F, shows that LL-37 was also found predominantly in the outer muscularis externa, as was RBD-3, and that the amount was dramatically reduced by I/R, while this loss was compensated for by a dramatic increase in the lower gland cells, in the area one would expect to find Paneth cells, of the mucosa.


Alterations in content and localization of defensins in rat ileum and jejunum following ischemia-reperfusion. Specific peptides, in specific places, for specific jobs?

Kozar RA, Santora RJ, Poindexter BJ, Milner SM, Bick RJ - Eplasty (2011)

This figure demonstrates the specificities with regard to both concentration changes and localizations of various antimicrobials in the ileum. Each pair of images (panels A through F) shows the sham tissue on the left and the ischemia-reperfusion image on the right. The first 2 panels (A) are images used to produce Figure 1. More rat beta defensin (RBD)-1 is seen in the apical and middle regions of the villi. Panel B again shows RBD-1 (red), but this time in the base/neck region of the villi. Panel C shows RBD-3 before and after ischemia-reperfusion demonstrating a decrease in peptide in the outer, longitudinal muscularis externa and a small increase in the inner, circular layer. Panel D shows RBD-2 increase in enterocytes due to I/R and a decrease in perilacteal concentration. Panel E shows RNP-1 throughout the epithelial layer of both sham and ischemia-reperfusion samples; panel F clearly demonstrates a reduction of cathelicidin in the outer, longitudinal muscularis externa, but an increase in the lower, glandular aspects of the mucosa (magnification X 400).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3044598&req=5

Figure 2: This figure demonstrates the specificities with regard to both concentration changes and localizations of various antimicrobials in the ileum. Each pair of images (panels A through F) shows the sham tissue on the left and the ischemia-reperfusion image on the right. The first 2 panels (A) are images used to produce Figure 1. More rat beta defensin (RBD)-1 is seen in the apical and middle regions of the villi. Panel B again shows RBD-1 (red), but this time in the base/neck region of the villi. Panel C shows RBD-3 before and after ischemia-reperfusion demonstrating a decrease in peptide in the outer, longitudinal muscularis externa and a small increase in the inner, circular layer. Panel D shows RBD-2 increase in enterocytes due to I/R and a decrease in perilacteal concentration. Panel E shows RNP-1 throughout the epithelial layer of both sham and ischemia-reperfusion samples; panel F clearly demonstrates a reduction of cathelicidin in the outer, longitudinal muscularis externa, but an increase in the lower, glandular aspects of the mucosa (magnification X 400).
Mentions: To further show the regulations and specificities, Figure 2 includes 6 pairs of images (shams on the left and I/R samples on the right). In each of the panels, the antimicrobial peptide is shown in red (orange/yellow where colocalization occurs). Cell nuclei are blue and F-actin is green. The first pair (panel A) are the images from Figure 1, but of note here is the high concentration of RBD-1 seen in the lower portion of the villi in the I/R sample. Panel B also shows RBD-1, but in the lower portion of the mucosa, where the crypts meet the glands. Note the increased amount of (red) peptide following I/R. Panel C is of RBD-3, located at an extremely high concentration in the longitudinal layer of the muscularis externa, but greatly reduced following I/R. However, this loss is somewhat compensated for by an increase in the inner, circular layer of the externa and maybe even in the myenteric plexus. Panel D demonstrates that I/R injury shows a trend of increasing RBD-2 in luminal epithelial cells in jejunum as seen in this pair of images, but not in ileum. Overall, the values are not significantly different; however, as stated, the trend within pairs is for an increase in staining intensity as shown here (13300 ± 7889 vs 16123 ± 7066; sham vs I/R pixels). Panel E shows that there was no change (18938 ± 8607 vs 18245 ± 7168; sham vs I/R; pixels) when comparing concentrations of RNP-1. The final pair of images, panel F, shows that LL-37 was also found predominantly in the outer muscularis externa, as was RBD-3, and that the amount was dramatically reduced by I/R, while this loss was compensated for by a dramatic increase in the lower gland cells, in the area one would expect to find Paneth cells, of the mucosa.

Bottom Line: Shams were subjected to laparotomy but no clamping.Ileum and jejunum were harvested and sectioned, and subjected to fluorescence deconvolution microscopy for determinations of content and localization of rat beta defensins, 1, 2, 3; rat neutrophil protein-1; and cathelicidin LL-37.Ischemia-reperfusion increased neutrophil defensin alpha (RNP-1) in jejunum; rat beta defensin 1 was increased 2-fold in ileal mucosa and slightly reduced in jejunal mucosa; rat beta defensin 2 was reduced by ischemia-reperfusion in ileum, but slightly increased in jejunum; rat beta defensin 3 was concentrated in the muscularis externa and myenteric plexus of the jejunum; ischemia-reperfusion did not alter cathelicidin LL-37 content in the small intestine, although a greater concentration was seen in jejunum compared with ileum.

View Article: PubMed Central - PubMed

ABSTRACT

Objective: To determine alterations in quantities and distributions of natural antimicrobials following ischemia-reperfusion injury. We hypothesized that these compounds would be upregulated in areas of small intestine where changes in permeability and cellular disruption were likely and where protective mechanisms would be initiated.

Methods: Rats with ischemia-reperfusion underwent superior mesenteric artery clamping and reperfusion. Shams were subjected to laparotomy but no clamping. Ileum and jejunum were harvested and sectioned, and subjected to fluorescence deconvolution microscopy for determinations of content and localization of rat beta defensins, 1, 2, 3; rat neutrophil protein-1; and cathelicidin LL-37. Modeling was performed to determine cellular location of antimicrobials.

Results: Ischemia-reperfusion increased neutrophil defensin alpha (RNP-1) in jejunum; rat beta defensin 1 was increased 2-fold in ileal mucosa and slightly reduced in jejunal mucosa; rat beta defensin 2 was reduced by ischemia-reperfusion in ileum, but slightly increased in jejunum; rat beta defensin 3 was concentrated in the muscularis externa and myenteric plexus of the jejunum; ischemia-reperfusion did not alter cathelicidin LL-37 content in the small intestine, although a greater concentration was seen in jejunum compared with ileum.

Conclusion: Ischemia-reperfusion injury caused changes in antimicrobial content in defined areas, and these different regulations might reflect the specific roles of jejunum versus ileum.

No MeSH data available.


Related in: MedlinePlus