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A clathrin light chain: a reporter mouse for in vivo imaging of endocytosis

Clathrin-mediated endocytosis (CME) is one of the best studied cellular uptake pathways and its contributions to nutrient uptake, receptor signaling, and maintenance of the lipid membrane homeostasis have been already elucidated. Today, we still have a lack of understanding how the different compone...

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Main Authors: Grimm, Elisabeth (Author) , Hoeven, Franciscus van der (Author) , Sardella, Donato (Author) , Willig, Katrin I. (Author) , Engel, Ulrike (Author) , Veits, Nisha (Author) , Engel, Robert (Author) , Cavalcanti-Adam, Elisabetta A. (Author) , Bestvater, Felix (Author) , Bordoni, Luca (Author) , Jennemann, Richard (Author) , Schönig, Kai (Author) , Schiessl, Ina Maria (Author) , Sandhoff, Roger (Author)
Format: Article (Journal)
Language:English
Published: September 23, 2022
In: PLOS ONE
Year: 2022, Volume: 17, Issue: 9, Pages: 1-24
ISSN:1932-6203
DOI:10.1371/journal.pone.0273660
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1371/journal.pone.0273660
Verlag, lizenzpflichtig, Volltext: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0273660
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Author Notes:Elisabeth Grimm, Franciscus van der Hoeven, Donato Sardella, Katrin I. Willig, Ulrike Engel, Nisha Veits, Robert Engel, Elisabetta Ada Cavalcanti-Adam, Felix Bestvater, Luca Bordoni, Richard Jennemann, Kai Schönig, Ina Maria Schiessl, Roger Sandhoff
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Summary:Clathrin-mediated endocytosis (CME) is one of the best studied cellular uptake pathways and its contributions to nutrient uptake, receptor signaling, and maintenance of the lipid membrane homeostasis have been already elucidated. Today, we still have a lack of understanding how the different components of this pathway cooperate dynamically in vivo. Therefore, we generated a reporter mouse model for CME by fusing eGFP endogenously in frame to clathrin light chain a (Clta) to track endocytosis in living mice. The fusion protein is expressed in all tissues, but in a cell specific manner, and can be visualized using fluorescence microscopy. Recruitment to nanobeads recorded by TIRF microscopy validated the functionality of the Clta-eGFP reporter. With this reporter model we were able to track the dynamics of Alexa594-BSA uptake in kidneys of anesthetized mice using intravital 2-photon microscopy. This reporter mouse model is not only a suitable and powerful tool to track CME in vivo in genetic or disease mouse models it can also help to shed light into the differential roles of the two clathrin light chain isoforms in health and disease.
Item Description:Gesehen am 19.04.2023
Physical Description:Online Resource
ISSN:1932-6203
DOI:10.1371/journal.pone.0273660

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