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Lineage-specific expression of bestrophin-2 and bestrophin-4 in human intestinal epithelial cells.

Ito G, Okamoto R, Murano T, Shimizu H, Fujii S, Nakata T, Mizutani T, Yui S, Akiyama-Morio J, Nemoto Y, Okada E, Araki A, Ohtsuka K, Tsuchiya K, Nakamura T, Watanabe M - PLoS ONE (2013)

Bottom Line: Consistently, the induction of goblet cell differentiation by a Notch inhibitor, LY411575, significantly up-regulated the expression of not BEST4 but BEST2 in MUC2-positive HT-29 cells.In addition, we found that the up- or down-regulation of Notch activity leads to the preferential expression of either BEST4 or BEST2, respectively, in LS174T cells.These results collectively confirmed that BEST2 and BEST4 could be added to the lineage-specific genes of humans IECs due to their abilities to clearly identify goblet cells of colonic origin and a distinct subset of absorptive cells, respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.

ABSTRACT
Intestinal epithelial cells (IECs) regulate the absorption and secretion of anions, such as HCO3(-) or Cl(-). Bestrophin genes represent a newly identified group of calcium-activated Cl(-) channels (CaCCs). Studies have suggested that, among the four human bestrophin-family genes, bestrophin-2 (BEST2) and bestrophin-4 (BEST4) might be expressed within the intestinal tissue. Consistently, a study showed that BEST2 is expressed by human colonic goblet cells. However, their precise expression pattern along the gastrointestinal tract, or the lineage specificity of the cells expressing these genes, remains largely unknown. Here, we show that BEST2 and BEST4 are expressed in vivo, each in a distinct, lineage-specific manner, in human IECs. While BEST2 was expressed exclusively in colonic goblet cells, BEST4 was expressed in the absorptive cells of both the small intestine and the colon. In addition, we found that BEST2 expression is significantly down-regulated in the active lesions of ulcerative colitis, where goblet cells were depleted, suggesting that BEST2 expression is restricted to goblet cells under both normal and pathologic conditions. Consistently, the induction of goblet cell differentiation by a Notch inhibitor, LY411575, significantly up-regulated the expression of not BEST4 but BEST2 in MUC2-positive HT-29 cells. Conversely, the induction of absorptive cell differentiation up-regulated the expression of BEST4 in villin-positive Caco-2 cells. In addition, we found that the up- or down-regulation of Notch activity leads to the preferential expression of either BEST4 or BEST2, respectively, in LS174T cells. These results collectively confirmed that BEST2 and BEST4 could be added to the lineage-specific genes of humans IECs due to their abilities to clearly identify goblet cells of colonic origin and a distinct subset of absorptive cells, respectively.

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Expression of BEST2 is markedly down-regulated in active lesions of UC.The immunohistochemical analysis of BEST2 and BEST4 expression using colonic tissues of UC patients is shown. Representative data obtained from three subjects in each group are presented. (A) Expression of BEST2 is down-regulated in the active lesions of UC patients, where goblet cell mucins are depleted. The double staining of BEST2 (green) and WGA-lectin (red) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). The red staining of WGA-lectin represents goblet cell mucins. Each series represents staining results obtained from a different patient (Original magnification 400x). (B) The expression of BEST4 is maintained at the surface epithelium of active, as well as inactive, lesions in UC patients. Single staining of BEST4 (green) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). Each figure represents a staining result obtained from a different patient.
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pone-0079693-g005: Expression of BEST2 is markedly down-regulated in active lesions of UC.The immunohistochemical analysis of BEST2 and BEST4 expression using colonic tissues of UC patients is shown. Representative data obtained from three subjects in each group are presented. (A) Expression of BEST2 is down-regulated in the active lesions of UC patients, where goblet cell mucins are depleted. The double staining of BEST2 (green) and WGA-lectin (red) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). The red staining of WGA-lectin represents goblet cell mucins. Each series represents staining results obtained from a different patient (Original magnification 400x). (B) The expression of BEST4 is maintained at the surface epithelium of active, as well as inactive, lesions in UC patients. Single staining of BEST4 (green) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). Each figure represents a staining result obtained from a different patient.

Mentions: We next asked whether the lineage-specific expression of BEST2 or BEST4 is also maintained under pathological conditions. The depletion of goblet cells is one of the unique pathologic changes that is observed in active lesions of UC [27]. Therefore, we examined whether BEST2-positive cells are depleted in the active lesions of UC. The analysis of colonic tissues obtained from the three patients showed that BEST2 expression is markedly down-regulated in the active lesions of UC in which goblet cells are depleted (Figure 5A). In contrast, the expression of BEST4 was maintained at almost the same level at the colonic surface epithelium of UC patients, irrespective of disease activity (Figure 5B). These results suggest that the expression of BEST2 is strictly regulated to goblet cells in both normal and pathological conditions. In addition, such a dramatic down-regulation of BEST2 in the active lesions of UC also suggests the possible involvement of BEST2 depletion in the pathophysiology of the disease.


Lineage-specific expression of bestrophin-2 and bestrophin-4 in human intestinal epithelial cells.

Ito G, Okamoto R, Murano T, Shimizu H, Fujii S, Nakata T, Mizutani T, Yui S, Akiyama-Morio J, Nemoto Y, Okada E, Araki A, Ohtsuka K, Tsuchiya K, Nakamura T, Watanabe M - PLoS ONE (2013)

Expression of BEST2 is markedly down-regulated in active lesions of UC.The immunohistochemical analysis of BEST2 and BEST4 expression using colonic tissues of UC patients is shown. Representative data obtained from three subjects in each group are presented. (A) Expression of BEST2 is down-regulated in the active lesions of UC patients, where goblet cell mucins are depleted. The double staining of BEST2 (green) and WGA-lectin (red) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). The red staining of WGA-lectin represents goblet cell mucins. Each series represents staining results obtained from a different patient (Original magnification 400x). (B) The expression of BEST4 is maintained at the surface epithelium of active, as well as inactive, lesions in UC patients. Single staining of BEST4 (green) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). Each figure represents a staining result obtained from a different patient.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818177&req=5

pone-0079693-g005: Expression of BEST2 is markedly down-regulated in active lesions of UC.The immunohistochemical analysis of BEST2 and BEST4 expression using colonic tissues of UC patients is shown. Representative data obtained from three subjects in each group are presented. (A) Expression of BEST2 is down-regulated in the active lesions of UC patients, where goblet cell mucins are depleted. The double staining of BEST2 (green) and WGA-lectin (red) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). The red staining of WGA-lectin represents goblet cell mucins. Each series represents staining results obtained from a different patient (Original magnification 400x). (B) The expression of BEST4 is maintained at the surface epithelium of active, as well as inactive, lesions in UC patients. Single staining of BEST4 (green) using tissues from the normal human colon or from active and inactive lesions of UC patients is shown (Original magnification 400x). Each figure represents a staining result obtained from a different patient.
Mentions: We next asked whether the lineage-specific expression of BEST2 or BEST4 is also maintained under pathological conditions. The depletion of goblet cells is one of the unique pathologic changes that is observed in active lesions of UC [27]. Therefore, we examined whether BEST2-positive cells are depleted in the active lesions of UC. The analysis of colonic tissues obtained from the three patients showed that BEST2 expression is markedly down-regulated in the active lesions of UC in which goblet cells are depleted (Figure 5A). In contrast, the expression of BEST4 was maintained at almost the same level at the colonic surface epithelium of UC patients, irrespective of disease activity (Figure 5B). These results suggest that the expression of BEST2 is strictly regulated to goblet cells in both normal and pathological conditions. In addition, such a dramatic down-regulation of BEST2 in the active lesions of UC also suggests the possible involvement of BEST2 depletion in the pathophysiology of the disease.

Bottom Line: Consistently, the induction of goblet cell differentiation by a Notch inhibitor, LY411575, significantly up-regulated the expression of not BEST4 but BEST2 in MUC2-positive HT-29 cells.In addition, we found that the up- or down-regulation of Notch activity leads to the preferential expression of either BEST4 or BEST2, respectively, in LS174T cells.These results collectively confirmed that BEST2 and BEST4 could be added to the lineage-specific genes of humans IECs due to their abilities to clearly identify goblet cells of colonic origin and a distinct subset of absorptive cells, respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.

ABSTRACT
Intestinal epithelial cells (IECs) regulate the absorption and secretion of anions, such as HCO3(-) or Cl(-). Bestrophin genes represent a newly identified group of calcium-activated Cl(-) channels (CaCCs). Studies have suggested that, among the four human bestrophin-family genes, bestrophin-2 (BEST2) and bestrophin-4 (BEST4) might be expressed within the intestinal tissue. Consistently, a study showed that BEST2 is expressed by human colonic goblet cells. However, their precise expression pattern along the gastrointestinal tract, or the lineage specificity of the cells expressing these genes, remains largely unknown. Here, we show that BEST2 and BEST4 are expressed in vivo, each in a distinct, lineage-specific manner, in human IECs. While BEST2 was expressed exclusively in colonic goblet cells, BEST4 was expressed in the absorptive cells of both the small intestine and the colon. In addition, we found that BEST2 expression is significantly down-regulated in the active lesions of ulcerative colitis, where goblet cells were depleted, suggesting that BEST2 expression is restricted to goblet cells under both normal and pathologic conditions. Consistently, the induction of goblet cell differentiation by a Notch inhibitor, LY411575, significantly up-regulated the expression of not BEST4 but BEST2 in MUC2-positive HT-29 cells. Conversely, the induction of absorptive cell differentiation up-regulated the expression of BEST4 in villin-positive Caco-2 cells. In addition, we found that the up- or down-regulation of Notch activity leads to the preferential expression of either BEST4 or BEST2, respectively, in LS174T cells. These results collectively confirmed that BEST2 and BEST4 could be added to the lineage-specific genes of humans IECs due to their abilities to clearly identify goblet cells of colonic origin and a distinct subset of absorptive cells, respectively.

Show MeSH
Related in: MedlinePlus