Cellular and sub-cellular localisation of oxyntomodulin-like immunoreactivity in enteroendocrine cells of human, mouse, pig and rat

Cellular and sub-cellular localisation of oxyntomodulin-like immunoreactivity in enteroendocrine cells of human, mouse, pig and rat

We use a monoclonal antibody against the C-terminal of oxyntomodulin (OXM) to investigate enteroendocrine cells (EEC) in mouse, rat, human and pig. This antibody has cross-reactivity with the OXM precursor, glicentin (Gli) but does not recognise glucagon. The antibody stained EEC in the jejunum and...

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Bibliographic Details
Published in:Cell and tissue research Vol. 375; no. 2; pp. 359 - 369
Main Authors: Fothergill, Linda J., Ringuet, Mitchell T., Sioras, Efstathia, Hunne, Billie, Fazio Coles, Therese E., Martins, Patricia R., Furness, John B.
Format: Journal Article
Language:English
Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-02-2019
Springer
Springer Nature B.V
Subjects:
Alternative right movement
Antibodies
Antigen-antibody reactions
Biomedical and Life Sciences
Biomedicine
Colon
Comparative analysis
Confederate symbols
Cross-reactivity
Enzyme-linked immunosorbent assay
Glucagon
Hormones
Human Genetics
Ileum
Immunoreactivity
Jejunum
Molecular Medicine
Monoclonal antibodies
Pancreas
Peptides
Proteomics
Regular Article
Secretory vesicles
Small intestine
Stomach
Human
Vesicular stores
Oxyntomodulin intestine
Gut hormones
Pancreas
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Summary:We use a monoclonal antibody against the C-terminal of oxyntomodulin (OXM) to investigate enteroendocrine cells (EEC) in mouse, rat, human and pig. This antibody has cross-reactivity with the OXM precursor, glicentin (Gli) but does not recognise glucagon. The antibody stained EEC in the jejunum and colon of each species. We compared OXM/Gli immunoreactivity with that revealed by antibodies against structurally related peptides, GLP-1 and glucagon and against GIP and PYY that are predicted to be in some EEC that express OXM/Gli. We used super-resolution to locate immunoreactive vesicles. In the pancreas, OXM/Gli was in glucagon cells but was located in separate storage vesicles to glucagon. In jejunal EEC, OXM/Gli and GIP were in many of the same cells but often in separate vesicles, whereas PYY and OXM/Gli were commonly colocalised in the same storage vesicles of colonic EEC. When binding of anti-GLP-1 to the structurally related GIP was removed by absorption with GIP peptide, GLP-1 and OXM/Gli immunoreactivities were contained in the same population of EEC in the intestine. We conclude that anti-OXM/Gli is a more reliable marker than anti-GLP-1 for EEC expressing preproglucagon products. Storage vesicles that were immunoreactive for OXM/Gli were almost always immunoreactive for GLP-1. OXM concentrations, measured by ELISA, were highest in the distal ileum and colon. Lesser concentrations were found in more proximal parts of small intestine and pancreas. Very little was in the stomach. In EEC containing GIP and OXM/Gli, these hormones are packaged in different secretory vesicles. Separate packaging also occurred for OXM and glucagon, whereas OXM/Gli and PYY and OXM/Gli and GLP-1 were commonly contained together in secretory vesicles.
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ISSN:0302-766X
1432-0878
DOI:10.1007/s00441-018-2921-z