Characterizing the Presence and Sensitivity of the P2X7 Receptor in Different Compartments of the Gut

Purinergic signaling has been established as an important feature of inflammation and homeostasis. The expression of a number of P2 receptor subtypes in the gut has been reported. In this study, using a well-known permeabilization method that is assessed by flow cytometry, we show that lymphocytes a...

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Bibliographic Details
Published in:Journal of innate immunity 2012-01, Vol.4 (5-6), p.529-541
Main Authors: de Campos, Nathalia Enes, Marques-da-Silva, Camila, Corrêa, Gladys, Castelo-Branco, Morgana Teixeira Lima, de Souza, Heitor Siffert Pereira, Coutinho-Silva, Robson
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - metabolism
Adenosine Triphosphate - pharmacology
Animals
ATP
Cations
Cell fate
Colon
Colon - cytology
Colon - metabolism
Digestive tract
Epithelial cells
Epithelial Cells - drug effects
Epithelial Cells - metabolism
Female
Flow cytometry
gamma -Interferon
Gastrointestinal Tract - cytology
Gastrointestinal Tract - metabolism
Gut-associated lymphoid tissues
Homeostasis
Inflammation
Intestine
Intestine, Small - cytology
Intestine, Small - metabolism
Lymph nodes
Lymph Nodes - cytology
Lymph Nodes - immunology
Lymph Nodes - metabolism
Lymphocytes
Lymphocytes - drug effects
Lymphocytes - immunology
Lymphocytes - metabolism
Macrophages
Macrophages - drug effects
Macrophages - immunology
Macrophages - metabolism
Male
Mesentery - metabolism
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Peritoneum
Peritoneum - cytology
Peritoneum - immunology
Peritoneum - metabolism
pH effects
Purine P2 receptors
Purine P2X receptors
Receptors, Purinergic P2X7 - metabolism
Regulatory sequences
Research Article
Signal Transduction - drug effects
Western blotting
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Summary:Purinergic signaling has been established as an important feature of inflammation and homeostasis. The expression of a number of P2 receptor subtypes in the gut has been reported. In this study, using a well-known permeabilization method that is assessed by flow cytometry, we show that lymphocytes and macrophages from the mesenteric lymph nodes (MLN) and the peritoneal cavity exhibit different sensitivities to extracellular ATP. Compared with the macrophages, the lymphocytes are more sensitive to ATP in the MLN compartment, whereas in the peritoneal cavity the macrophages are more sensitive to ATP than the lymphocytes. In addition, we have shown that the epithelial cells from the small bowel are more resistant to the ATP effects than the cells from the colon. These cells, however, become susceptible after exposure to IFN-γ. Furthermore, by examining parameters such as pH manipulation, the exposure to divalent cations and the P2X7 antagonist Brilliant Blue G, and the use of cells from P2X7 –/– mice, we have shown that the P2X7 receptors are the ATP-activated receptors responsible for the permeabilization phenomenon. In addition, using Western blot analysis, we have demonstrated the changes in the P2X7 receptor expression in immune cells isolated from different sites in the gut and in the gut-associated lymphoid tissues. Our findings suggest the existence of the site-specific modulation of P2X7 receptors on epithelial and immune cells, and we define purinergic signaling as a new regulatory element in the control of inflammation and cell fate in the gut and in the gut-associated lymphoid tissues.
ISSN:1662-811X
1662-8128
DOI:10.1159/000336628