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


Journal

PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081

Informations de publication

Date de publication:
2022
Historique:
received: 16 12 2021
accepted: 13 08 2022
entrez: 23 9 2022
pubmed: 24 9 2022
medline: 28 9 2022
Statut: epublish

Résumé

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.

Identifiants

pubmed: 36149863
doi: 10.1371/journal.pone.0273660
pii: PONE-D-21-39683
pmc: PMC9506643
doi:

Substances chimiques

Clathrin 0
Clathrin Light Chains 0
Lipids 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e0273660

Déclaration de conflit d'intérêts

The authors have declared that no competing interests exist.

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Auteurs

Elisabeth Grimm (E)

Lipid Pathobiochemistry Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Faculty of Biosciences, Heidelberg University, Heidelberg, Germany.

Franciscus van der Hoeven (F)

Transgenic Service, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Donato Sardella (D)

Department of Biomedicine, Aarhus University, Aarhus, Denmark.

Katrin I Willig (KI)

Optical Nanoscopy in Neuroscience, Center for Nanoscale Microscopy and Molecular Physiology of the Brain, University Medical Center Göttingen, Göttingen, Germany.
Max Planck Institute of Experimental Medicine, Goettingen, Germany.

Ulrike Engel (U)

Faculty of Biosciences, Heidelberg University, Heidelberg, Germany.
Nikon Imaging Center at Heidelberg University and Centre of Organismal Studies (COS), Bioquant, Heidelberg, Germany.

Nisha Veits (N)

Department of Cellular Biophysics, Max Planck Institute for Medical Research, Heidelberg, Germany.

Robert Engel (R)

Lipid Pathobiochemistry Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Faculty of Biosciences, Heidelberg University, Heidelberg, Germany.

Elisabetta Ada Cavalcanti-Adam (EA)

Department of Cellular Biophysics, Max Planck Institute for Medical Research, Heidelberg, Germany.

Felix Bestvater (F)

Light Microscopy Core Facility, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Luca Bordoni (L)

Department of Biomedicine, Aarhus University, Aarhus, Denmark.

Richard Jennemann (R)

Lipid Pathobiochemistry Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Kai Schönig (K)

Department of Molecular Biology, Central Institute of Mental Health, Mannheim, Germany.

Ina Maria Schiessl (IM)

Department of Biomedicine, Aarhus University, Aarhus, Denmark.

Roger Sandhoff (R)

Lipid Pathobiochemistry Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.

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Classifications MeSH