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Expression of Bis in the mouse gastrointestinal system.

Lee YD, Yoon JS, Yoon HH, Youn HJ, Kim J, Lee JH - Anat Cell Biol (2012)

Bottom Line: Ganglionated plexuses, located in submucous layers, as well as intermuscular layers, were specifically immunoreactive for Bis.Immunostaining with neuron specific esterase antibodies indicate that Bis is also present in the cell bodies of ganglions in the enteric nervous system (ENS).Our findings indicate that Bis plays a role in regulating GI functions, such as motility and absorption, through modulating signal transmission between the ENS and smooth muscles or the intestinal epitheliums.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, The Catholic University of Korea College of Medicine, Seoul, Korea.

ABSTRACT
The Bcl-2 interacting death suppressor (Bis) protein is known to be involved in a variety of pathophysiological conditions. We recently generated bis-deficient mice, which exhibited early lethality with typical nutritional deprivation status. To further investigate the molecular basis for the malnutrition phenotype of bis deficient mice, we explored Bis expression in the digestive system of normal mice. Western blot analysis and quantitative real time reverse transcription polymerase chain reaction analysis indicated that Bis expression is highest in the esophagus, followed by the stomach, colon, jejunum and ileum. Immunohistochemical data indicated that Bis expression is restricted to the stratified squamous epitheliums in the esophagus and forestomach, and was not notable in the columnar epitheliums in the stomach, small intestine and colon. In addition, strong Bis immunoreactivity was detected in the striated muscles surrounding the esophagus and smooth muscles at a lesser intensity throughout the gastrointestinal (GI) tract. Ganglionated plexuses, located in submucous layers, as well as intermuscular layers, were specifically immunoreactive for Bis. Immunofluorescence studies revealed that Bis is co-localized in glial fibrillary acidic protein-expressing enteric glial cells. Immunostaining with neuron specific esterase antibodies indicate that Bis is also present in the cell bodies of ganglions in the enteric nervous system (ENS). Our findings indicate that Bis plays a role in regulating GI functions, such as motility and absorption, through modulating signal transmission between the ENS and smooth muscles or the intestinal epitheliums.

No MeSH data available.


Related in: MedlinePlus

Expression of Bis in the mouse gastrointestinal (GI) tract. (A) Western blot analysis for Bis in the each part of GI tracts from 14-day-old mice. A representative result was provided with heat shock protein 70 (Hsp70) as a loading control. (B) A quantitative determination of Bis expression using densitometric analysis after normalizing to Hsp70 (mean±SE, n=3). The relative density of the Bis bands to that of Hsp70 from ileum was designated as 1.0. (C) Determination of the relative expression of bis mRNA in the mouse GI tract by quantitative real time polymerase chain reaction analysis. Data are normalized relative to β-actin mRNA in the same samples, and the value for the ileum was arbitrarily set as 1.0 (mean±SE, n=3). S.M., skeletal muscle; ESO, esophagus; STO, stomach; JEJ, jejunum; ILE, ileum; COL, colon. *P<0.05, **P<0.01 and ***P<0.005 compared with the value from ileum.
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Figure 1: Expression of Bis in the mouse gastrointestinal (GI) tract. (A) Western blot analysis for Bis in the each part of GI tracts from 14-day-old mice. A representative result was provided with heat shock protein 70 (Hsp70) as a loading control. (B) A quantitative determination of Bis expression using densitometric analysis after normalizing to Hsp70 (mean±SE, n=3). The relative density of the Bis bands to that of Hsp70 from ileum was designated as 1.0. (C) Determination of the relative expression of bis mRNA in the mouse GI tract by quantitative real time polymerase chain reaction analysis. Data are normalized relative to β-actin mRNA in the same samples, and the value for the ileum was arbitrarily set as 1.0 (mean±SE, n=3). S.M., skeletal muscle; ESO, esophagus; STO, stomach; JEJ, jejunum; ILE, ileum; COL, colon. *P<0.05, **P<0.01 and ***P<0.005 compared with the value from ileum.

Mentions: Tissue fractions were isolated from each part of the GI tract of mice at postnatal 14-16 days, and prepared for western analysis, quantitative real time analysis and immunohistochemistry. Western analysis, using specific Bis antibody, demonstrated that the degree for Bis expression varied throughout the GI tract, although the expression levels were lower than those of the skeletal muscles (Fig. 1A). Among the GI tract, the highest expression of Bis was by the esophagus, while only traces were found for the jejunum and ileum. The quantitative determination of band intensities indicated that Bis expression in the esophagus was 77 fold higher than that for the ileum (Fig. 1B). The relative bis mRNA levels were the highest in the esophagus, 38 fold higher than that for the jejunum or ileum, which is correlated with the Bis protein expression pattern in a Western assay (Fig. 1C).


Expression of Bis in the mouse gastrointestinal system.

Lee YD, Yoon JS, Yoon HH, Youn HJ, Kim J, Lee JH - Anat Cell Biol (2012)

Expression of Bis in the mouse gastrointestinal (GI) tract. (A) Western blot analysis for Bis in the each part of GI tracts from 14-day-old mice. A representative result was provided with heat shock protein 70 (Hsp70) as a loading control. (B) A quantitative determination of Bis expression using densitometric analysis after normalizing to Hsp70 (mean±SE, n=3). The relative density of the Bis bands to that of Hsp70 from ileum was designated as 1.0. (C) Determination of the relative expression of bis mRNA in the mouse GI tract by quantitative real time polymerase chain reaction analysis. Data are normalized relative to β-actin mRNA in the same samples, and the value for the ileum was arbitrarily set as 1.0 (mean±SE, n=3). S.M., skeletal muscle; ESO, esophagus; STO, stomach; JEJ, jejunum; ILE, ileum; COL, colon. *P<0.05, **P<0.01 and ***P<0.005 compared with the value from ileum.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3472142&req=5

Figure 1: Expression of Bis in the mouse gastrointestinal (GI) tract. (A) Western blot analysis for Bis in the each part of GI tracts from 14-day-old mice. A representative result was provided with heat shock protein 70 (Hsp70) as a loading control. (B) A quantitative determination of Bis expression using densitometric analysis after normalizing to Hsp70 (mean±SE, n=3). The relative density of the Bis bands to that of Hsp70 from ileum was designated as 1.0. (C) Determination of the relative expression of bis mRNA in the mouse GI tract by quantitative real time polymerase chain reaction analysis. Data are normalized relative to β-actin mRNA in the same samples, and the value for the ileum was arbitrarily set as 1.0 (mean±SE, n=3). S.M., skeletal muscle; ESO, esophagus; STO, stomach; JEJ, jejunum; ILE, ileum; COL, colon. *P<0.05, **P<0.01 and ***P<0.005 compared with the value from ileum.
Mentions: Tissue fractions were isolated from each part of the GI tract of mice at postnatal 14-16 days, and prepared for western analysis, quantitative real time analysis and immunohistochemistry. Western analysis, using specific Bis antibody, demonstrated that the degree for Bis expression varied throughout the GI tract, although the expression levels were lower than those of the skeletal muscles (Fig. 1A). Among the GI tract, the highest expression of Bis was by the esophagus, while only traces were found for the jejunum and ileum. The quantitative determination of band intensities indicated that Bis expression in the esophagus was 77 fold higher than that for the ileum (Fig. 1B). The relative bis mRNA levels were the highest in the esophagus, 38 fold higher than that for the jejunum or ileum, which is correlated with the Bis protein expression pattern in a Western assay (Fig. 1C).

Bottom Line: Ganglionated plexuses, located in submucous layers, as well as intermuscular layers, were specifically immunoreactive for Bis.Immunostaining with neuron specific esterase antibodies indicate that Bis is also present in the cell bodies of ganglions in the enteric nervous system (ENS).Our findings indicate that Bis plays a role in regulating GI functions, such as motility and absorption, through modulating signal transmission between the ENS and smooth muscles or the intestinal epitheliums.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, The Catholic University of Korea College of Medicine, Seoul, Korea.

ABSTRACT
The Bcl-2 interacting death suppressor (Bis) protein is known to be involved in a variety of pathophysiological conditions. We recently generated bis-deficient mice, which exhibited early lethality with typical nutritional deprivation status. To further investigate the molecular basis for the malnutrition phenotype of bis deficient mice, we explored Bis expression in the digestive system of normal mice. Western blot analysis and quantitative real time reverse transcription polymerase chain reaction analysis indicated that Bis expression is highest in the esophagus, followed by the stomach, colon, jejunum and ileum. Immunohistochemical data indicated that Bis expression is restricted to the stratified squamous epitheliums in the esophagus and forestomach, and was not notable in the columnar epitheliums in the stomach, small intestine and colon. In addition, strong Bis immunoreactivity was detected in the striated muscles surrounding the esophagus and smooth muscles at a lesser intensity throughout the gastrointestinal (GI) tract. Ganglionated plexuses, located in submucous layers, as well as intermuscular layers, were specifically immunoreactive for Bis. Immunofluorescence studies revealed that Bis is co-localized in glial fibrillary acidic protein-expressing enteric glial cells. Immunostaining with neuron specific esterase antibodies indicate that Bis is also present in the cell bodies of ganglions in the enteric nervous system (ENS). Our findings indicate that Bis plays a role in regulating GI functions, such as motility and absorption, through modulating signal transmission between the ENS and smooth muscles or the intestinal epitheliums.

No MeSH data available.


Related in: MedlinePlus