Universität zu Köln

Wiesner, Eva ORCID: 0000-0002-4096-6374, Binz-Lotter, Julia ORCID: 0000-0003-4678-0703, Hackl, Agnes, Unnersjö-Jess, David ORCID: 0000-0002-4162-0973, Rutkowski, Nelli ORCID: 0000-0003-3672-4310, Benzing, Thomas ORCID: 0000-0003-0512-1066 and Hackl, Matthias J. ORCID: 0000-0002-4199-3374 (2024). Correlative multiphoton-STED microscopy of podocyte calcium levels and slit diaphragm ultrastructure in the renal glomerulus. Scientific Reports, 14 (1). pp. 1-10. Springer Nature. ISSN 2045-2322

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Abstract

[Artikel-Nr.: 13019] 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.

Item Type:	Article
Creators:	Creators Email ORCID ORCID Put Code Wiesner, Eva UNSPECIFIED https://orcid.org/0000-0002-4096-6374 UNSPECIFIED Binz-Lotter, Julia UNSPECIFIED https://orcid.org/0000-0003-4678-0703 UNSPECIFIED Hackl, Agnes UNSPECIFIED UNSPECIFIED UNSPECIFIED Unnersjö-Jess, David UNSPECIFIED https://orcid.org/0000-0002-4162-0973 UNSPECIFIED Rutkowski, Nelli UNSPECIFIED https://orcid.org/0000-0003-3672-4310 UNSPECIFIED Benzing, Thomas UNSPECIFIED https://orcid.org/0000-0003-0512-1066 UNSPECIFIED Hackl, Matthias J. UNSPECIFIED https://orcid.org/0000-0002-4199-3374 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-815603
Identification Number:	10.1038/s41598-024-63507-9
Journal or Publication Title:	Scientific Reports
Volume:	14
Number:	1
Page Range:	pp. 1-10
Number of Pages:	10
Date:	6 June 2024
Publisher:	Springer Nature
ISSN:	2045-2322
Language:	English
Faculty:	Faculty of Medicine
Divisions:	CECAD - Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases Faculty of Medicine > Innere Medizin > Klinik II für Innere Medizin - Nephrologie, Rheumatologie, Diabetologie und Allgemeine Innere Medizin Faculty of Medicine > Kinder- und Jugendmedizin > Klinik und Poliklinik für Kinder- und Jugendmedizin Zentrum für Molekulare Medizin
Subjects:	Medical sciences Medicine
['eprint_fieldname_oa_funders' not defined]:	Publikationsfonds UzK
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/81560