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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Published in:PloS one 2022-09, Vol.17 (9), p.e0273660-e0273660
Main Authors: Grimm, Elisabeth, van der Hoeven, Franciscus, Sardella, Donato, Willig, Katrin I, Engel, Ulrike, Veits, Nisha, Engel, Robert, Cavalcanti-Adam, Elisabetta Ada, Bestvater, Felix, Bordoni, Luca, Jennemann, Richard, Schönig, Kai, Schiessl, Ina Maria, Sandhoff, Roger
Format: Article
Language:English
Subjects:
Animal models
Animal models in research
Animals
Binding sites
Biology and Life Sciences
Care and treatment
Chains
Clathrin
CRISPR
Diagnosis
Endocytosis
Fluorescence
Fluorescence microscopy
Fusion protein
Genetic aspects
Health aspects
Homeostasis
Isoforms
Kidneys
Lipids
Medicine and Health Sciences
Methods
Microscopy
Modelling
Mutation
Nutrient uptake
Physiology
Plasma
Proteins
Receptor-mediated endocytosis
Research and Analysis Methods
Small intestine
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cited_by cdi_FETCH-LOGICAL-c669t-f2238996ab2afd3d4d18a05bb133efbdc488142337476310237707f7cf15870f3
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creator Grimm, Elisabeth
van der Hoeven, Franciscus
Sardella, Donato
Willig, Katrin I
Engel, Ulrike
Veits, Nisha
Engel, Robert
Cavalcanti-Adam, Elisabetta Ada
Bestvater, Felix
Bordoni, Luca
Jennemann, Richard
Schönig, Kai
Schiessl, Ina Maria
Sandhoff, Roger
description 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.
doi_str_mv 10.1371/journal.pone.0273660
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1932-6203
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source Open Access: PubMed Central; Publicly Available Content (ProQuest); Coronavirus Research Database
subjects Animal models
Animal models in research
Animals
Binding sites
Biology and Life Sciences
Care and treatment
Chains
Clathrin
CRISPR
Diagnosis
Endocytosis
Fluorescence
Fluorescence microscopy
Fusion protein
Genetic aspects
Health aspects
Homeostasis
Isoforms
Kidneys
Lipids
Medicine and Health Sciences
Methods
Microscopy
Modelling
Mutation
Nutrient uptake
Physiology
Plasma
Proteins
Receptor-mediated endocytosis
Research and Analysis Methods
Small intestine
title A Clathrin light chain A reporter mouse for in vivo imaging of endocytosis
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