A Clathrin light chain A reporter mouse for in vivo imaging of endocytosis

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

doi:10.1371/journal.pone.0273660

A Clathrin light chain A reporter mouse for in vivo imaging of endocytosis

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^*

^*Corresponding author for this work

Department of Biomedicine - Forskning og uddannelse, Syd

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

4 Citations (Scopus)

Abstract

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.

Original language	English
Article number	e0273660
Journal	PLOS ONE
Volume	17
Issue	9
Number of pages	24
ISSN	1932-6203
DOIs	https://doi.org/10.1371/journal.pone.0273660
Publication status	Published - Sept 2022

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title = "A Clathrin light chain A reporter mouse for in vivo imaging of endocytosis",

abstract = "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.",

author = "Elisabeth Grimm and {van der Hoeven}, Franciscus and Donato Sardella and Willig, {Katrin I} and Ulrike Engel and Nisha Veits and Robert Engel and Cavalcanti-Adam, {Elisabetta Ada} and Felix Bestvater and Luca Bordoni and Richard Jennemann and Kai Sch{\"o}nig and Schiessl, {Ina Maria} and Roger Sandhoff",

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AU - Grimm, Elisabeth

AU - van der Hoeven, Franciscus

AU - Sardella, Donato

AU - Willig, Katrin I

AU - Engel, Ulrike

AU - Veits, Nisha

AU - Engel, Robert

AU - Cavalcanti-Adam, Elisabetta Ada

AU - Bestvater, Felix

AU - Bordoni, Luca

AU - Jennemann, Richard

AU - Schönig, Kai

AU - Schiessl, Ina Maria

AU - Sandhoff, Roger

PY - 2022/9

Y1 - 2022/9

N2 - 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.

AB - 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.

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JO - PLOS ONE

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A Clathrin light chain A reporter mouse for in vivo imaging of endocytosis

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