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Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids
Published on: December 13, 2017
Electron microscopic definition of intestinal endocrine cells: immunogold localization and review
1Department of Internal Medicine, University of Michigan Medical School, Ann Arbor.
Insights
Identifying intestinal endocrine cells requires specific staining, as morphology alone is often insufficient. While some cell types show distinct granules, absolute identification relies on immunogold staining for accurate classification.
Area of Science:
- Gastroenterology
- Cell Biology
- Histology
Background:
- Intestinal endocrine cells play crucial roles in regulating physiological functions.
- Accurate identification of these cells is essential for understanding their functions and associated pathologies.
- Previous studies relied heavily on morphological criteria, with limited success in distinguishing all cell subtypes.
Purpose of the Study:
- To evaluate the reliability of morphological criteria versus specific immunostaining for identifying intestinal endocrine cells.
- To determine which endocrine cell types can be definitively identified by ultrastructural morphology alone.
- To establish definitive identification methods for all intestinal enteroendocrine cell subtypes.
Main Methods:
- Analysis of intestinal mucosal biopsy specimens over 25 years.
- Utilized electron microscopy for ultrastructural examination.
- Employed specific antisera and immunogold staining for cell characterization.
Main Results:
- Morphological criteria were insufficient for absolute identification of most intestinal endocrine cells.
- D, G, EC1, EC2, ECn, D1, and intestinal gastrin cells showed characteristic granules, aiding identification.
- Absolute identification of D, G, IG, and TG cells requires specific antisera.
- I, L, N, and K cells are morphologically similar and necessitate specific staining.
- Mo, S, and P cells were not identified by either method, though P cells may resemble D1 cells.
Conclusions:
- Absolute identification of intestinal enteroendocrine cells using electron microscopy necessitates specific immunostaining.
- Morphological assessment is useful for certain cell types (D, G, EC1, EC2, ECn, D1, IG) when granules are abundant.
- Further research is needed to identify specific markers for Mo, S, and P cells.
Abstract:
Intestinal mucosal biopsy specimens processed during the past 25 years were used to example the ultrastructural characteristics of intestinal endocrine cells. The cells were defined on the basis of morphologic criteria and, when feasible, with specific antisera and immunogold staining. The hypothesis was that each endocrine cell, once well defined, should be identifiable on the basis of standard morphologic criteria not requiring specific immunostaining. This was not the case, D, G, EC1, EC2, ECn, D1, and intestinal gastrin cells have characteristic secretory granules and, when sufficient granules are present, can be identified consistently on the basis of morphologic criteria. Absolute identification of D, G, IG, and TG cells requires staining with specific antisera, a condition easily obtainable only for D, G, and IG cells. D1, EC1, EC2, and ECn cells must be identified morphologically until secretory products specific for each of these cells are identified. I, L, N, and K cells are remarkably similar in appearance and must be distinguished by specific staining. Mo, S, and P cells were not identified by either morphologic appearance or immunostaining. It is suggested that a cell similar to the D1 cell but with exceptionally small granules may be the P cell. Absolute identification of intestinal enteroendocrine cells by electron microscopy requires specific staining. The characteristic appearance of the secretory granules of many of these cells (D, G, EC1, EC2, ECn, D1, and IG) permits morphologic identification when numerous secretory granules are present.
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