Correlative multiphoton-STED microscopy of podocyte calcium levels and slit diaphragm ultrastructure in the renal glomerulus.

Animals Podocytes / metabolism Calcium / metabolism Mice Kidney Glomerulus / metabolism Microscopy, Fluorescence, Multiphoton / methods

Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
06 06 2024
Historique:
received: 23 02 2024
accepted: 29 05 2024
medline: 7 6 2024
pubmed: 7 6 2024
entrez: 6 6 2024
Statut: epublish

Résumé

In recent years functional multiphoton (MP) imaging of vital mouse tissues and stimulation emission depletion (STED) imaging of optically cleared tissues allowed new insights into kidney biology. Here, we present a novel workflow where MP imaging of calcium signals can be combined with super-resolved STED imaging for morphological analysis of the slit diaphragm (SD) within the same glomerulus. Mice expressing the calcium indicator GCaMP3 in podocytes served as healthy controls or were challenged with two different doses of nephrotoxic serum (NTS). NTS induced glomerular damage in a dose dependent manner measured by shortening of SD length. In acute kidney slices (AKS) intracellular calcium levels increased upon disease but showed a high variation between glomeruli. We could not find a clear correlation between intracellular calcium levels and SD length in the same glomerulus. Remarkably, analysis of the SD morphology of glomeruli selected during MP calcium imaging revealed a higher percentage of completely disrupted SD architecture than estimated by STED imaging alone. Our novel co-imaging protocol is applicable to a broad range of research questions. It can be used with different tissues and is compatible with diverse reporters and target proteins.

Identifiants

DOI: 10.1038/s41598-024-63507-9 PMID: 38844492
pubmed: 38844492
doi: 10.1038/s41598-024-63507-9
pii: 10.1038/s41598-024-63507-9
doi:

Substances chimiques

Calcium SY7Q814VUP

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

13019

Subventions

Organisme : Deutsche Forschungsgemeinschaft
ID : 360043781
Organisme : Deutsche Forschungsgemeinschaft
ID : HA6212/3-1

Informations de copyright

© 2024. The Author(s).

Références

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Auteurs

Eva Wiesner (E)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.

Julia Binz-Lotter (J)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.

Agnes Hackl (A)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Department of Pediatrics, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.

David Unnersjö-Jess (D)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.

Nelli Rutkowski (N)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.

Thomas Benzing (T)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.

Matthias J Hackl (MJ)

Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany. Matthias.hackl@uk-koeln.de.
Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany. Matthias.hackl@uk-koeln.de.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Correlative multiphoton-STED microscopy of...
questionsmedicales.fr › Cellules › Cellules épithéliales › Podocytes › Correlative multiphoton-STED microscopy of...
questionsmedicales.fr › Facteurs biologiques › Facteurs de la coagulation sanguine › Calcium › Correlative multiphoton-STED microscopy of...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › Correlative multiphoton-STED microscopy of...
questionsmedicales.fr › Appareil urogénital › Voies urinaires › Rein › Néphrons › Glomérule rénal › Correlative multiphoton-STED microscopy of...
questionsmedicales.fr › Techniques d'investigation › Microscopie optique non linéaire › Microscopie de fluorescence multiphotonique › Correlative multiphoton-STED microscopy of...