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. 2000 Oct;47(4):481-6.
doi: 10.1136/gut.47.4.481.

Expression of leptin and leptin receptor isoforms in the human stomach

H Mix  1 , A Widjaja, O Jandl, M Cornberg, A Kaul, M Göke, W Beil, M Kuske, G Brabant, M P Manns, S Wagner
Affiliations

Expression of leptin and leptin receptor isoforms in the human stomach

H Mix et al. Gut. 2000 Oct.

Abstract

Background: Leptin is an important regulator of food intake and energy expenditure. Initially it was thought to be expressed exclusively in and secreted by adipocytes. Recently, leptin expression was also noted in other tissues, including rat gastric mucosa. Information on leptin and leptin receptor expression in the human stomach is lacking.

Aim: To investigate expression of leptin and its corresponding receptors in human gastric epithelial cells.

Methods: Fundic and antral gastric mucosal biopsies, primary cultures of human gastric epithelial cells, and the human gastric cancer cell line AGS were screened for expression of leptin and different leptin receptor isoform mRNA by reverse transcriptase-polymerase chain reaction. Immunohistochemistry was performed for localisation of leptin and leptin receptor proteins in gastric mucosa.

Results: mRNA of leptin and its four receptor isoforms (huOB-R, long receptor isoform; huB219.1-3, short receptor isoforms) was detected in gastric mucosal biopsies, cultured human gastric epithelial cells, and gastric cancer cells. Immunohistochemistry demonstrated that chief as well as parietal cells were reactive to leptin and leptin receptors.

Conclusions: Leptin and leptin receptors are expressed in human gastric mucosa. These findings suggest a paracrine and/or autocrine effect of leptin on gastric epithelial cell function.

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Figures

Figure 1
Figure 1
Detection of leptin mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR). Lanes 1+2, adipose tissue; lanes 3+4, human gastric mucosal biopsy; lanes 5+6, primary cultured human gastric epithelial cells; lanes 7+8, human cancer cell line AGS. The PCR products of leptin specific mRNA are demonstrated in lanes 2, 4, 6, and 8. Lanes 1, 3, 5, and 7 show the house keeping gene GAPDH. M, size marker 100 bp DNA ladder.
Figure 2
Figure 2
Detection of different leptin receptor mRNA splice variants by reverse transcriptase-polymerase chain reaction (RT-PCR). (A) Human gastric mucosal biopsy. (B) Primary cultured human gastric epithelial cells. (C) Human cancer cell line AGS. huOB-R, human long leptin receptor isoform; huB219.1-huB219.3, human short leptin receptor isoforms, huOB-Runi, primers chosen to detect all receptor isoforms.
Figure 3
Figure 3
Immunohistochemical detection of leptin (A, C) and pepsinogen (B, D) in serial sections of human fundic mucosal biopsies. (A, B) Overview (original magnification ×100). (C, D) Detailed view (original magnification ×400). Arrows denote corresponding cells of adjacent sections which are leptin positive and pepsinogen negative. Paraffin embedded, formalin fixed tissue sections were analysed using a streptavidin-peroxidase technique as described under materials and methods.
Figure 4
Figure 4
Double immunostaining for leptin (A) and parietal cell antigen (B) in human fundic gastric mucosa (original magnification ×1000). Paraffin embedded, formalin fixed tissue sections were analysed using a streptavidin-peroxidase technique (A) and FITC labelled immunofluorescence (B) as described in materials and methods.
Figure 5
Figure 5
(A, B) Double immunofluorescent staining for pepsinogen (A, green) and leptin receptor (B, red) in tissue sections of human fundic gastric mucosa (original magnification ×400). Chief cells were identified by rabbit polyclonal antipepsinogen antibody (dilution 1:125) and FITC conjugated goat antirabbit IgG (dilution 1:100) in cryostat sections. Subsequently mouse monoclonal antileptin receptor IgG1 (B-3, dilution 1:50) and rhodamine conjugated goat antimouse IgG (dilution 1:100) were used. (C, D) Double immunofluorescent staining for parietal cell antigen (C, green) and leptin receptor (D, red) in tissue sections of human fundic gastric mucosa (original magnification ×400). Parietal cells were identified by serum of an antiparietal cell positive patient (dilution 1:80) and FITC conjugated goat anti-human IgG (dilution 1:50) in cryostat sections. Subsequently rabbit polyclonal antileptin receptor antibody (H-300, dilution 1:50) and rhodamine conjugated goat antirabbit IgG (dilution 1:75) were used.
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