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An Approach to Intersectionally Target Mature Enteroendocrine Cells in the Small Intestine of Mice

Authors :
Christian Vossen
Patricia Schmidt
Claudia Maria Wunderlich
Melanie Joyce Mittenbühler
Claas Tapken
Peter Wienand
Paul Nicolas Mirabella
Leonie Cabot
Anna-Lena Schumacher
Kat Folz-Donahue
Christian Kukat
Ingo Voigt
Jens C. Brüning
Henning Fenselau
F. Thomas Wunderlich
Source :
Cells, Vol 13, Iss 1, p 102 (2024)
Publication Year :
2024
Publisher :
MDPI AG, 2024.

Abstract

Enteroendocrine cells (EECs) constitute only a small proportion of Villin-1 (Vil1)-expressing intestinal epithelial cells (IECs) of the gastrointestinal tract; yet, in sum, they build the largest endocrine organ of the body, with each of them storing and releasing a distinct set of peptides for the control of feeding behavior, glucose metabolism, and gastrointestinal motility. Like all IEC types, EECs are continuously renewed from intestinal stem cells in the crypt base and terminally differentiate into mature subtypes while moving up the crypt–villus axis. Interestingly, EECs adjust their hormonal secretion according to their migration state as EECs receive altering differentiation signals along the crypt–villus axis and thus undergo functional readaptation. Cell-specific targeting of mature EEC subtypes by specific promoters is challenging because the expression of EEC-derived peptides and their precursors is not limited to EECs but are also found in other organs, such as the brain (e.g., Cck and Sst) as well as in the pancreas (e.g., Sst and Gcg). Here, we describe an intersectional genetic approach that enables cell type-specific targeting of functionally distinct EEC subtypes by combining a newly generated Dre-recombinase expressing mouse line (Vil1-2A-DD-Dre) with multiple existing Cre-recombinase mice and mouse strains with rox and loxP sites flanked stop cassettes for transgene expression. We found that transgene expression in triple-transgenic mice is highly specific in I but not D and L cells in the terminal villi of the small intestine. The targeting of EECs only in terminal villi is due to the integration of a defective 2A separating peptide that, combined with low EEC intrinsic Vil1 expression, restricts our Vil1-2A-DD-Dre mouse line and the intersectional genetic approach described here only applicable for the investigation of mature EEC subpopulations.

Details

Language :
English
ISSN :
20734409
Volume :
13
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Cells
Publication Type :
Academic Journal
Accession number :
edsdoj.f10d02ed844a188e6cca6ae65c46b1
Document Type :
article
Full Text :
https://doi.org/10.3390/cells13010102