دورية أكاديمية
Correlative multiphoton-STED microscopy of podocyte calcium levels and slit diaphragm ultrastructure in the renal glomerulus
العنوان: | Correlative multiphoton-STED microscopy of podocyte calcium levels and slit diaphragm ultrastructure in the renal glomerulus |
---|---|
المؤلفون: | Eva Wiesner, Julia Binz-Lotter, Agnes Hackl, David Unnersjö-Jess, Nelli Rutkowski, Thomas Benzing, Matthias J. Hackl |
المصدر: | Scientific Reports, Vol 14, Iss 1, Pp 1-10 (2024) |
بيانات النشر: | Nature Portfolio, 2024. |
سنة النشر: | 2024 |
المجموعة: | LCC:Medicine LCC:Science |
مصطلحات موضوعية: | Medicine, Science |
الوصف: | Abstract 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. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English |
تدمد: | 2045-2322 |
Relation: | https://doaj.org/toc/2045-2322 |
DOI: | 10.1038/s41598-024-63507-9 |
URL الوصول: | https://doaj.org/article/7f01487f736845bebb0684e500ef792b |
رقم الأكسشن: | edsdoj.7f01487f736845bebb0684e500ef792b |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 20452322 |
---|---|
DOI: | 10.1038/s41598-024-63507-9 |