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Temporal calcium activity in metanephric mesenchyme cells regulates kidney branching morphogenesis
Karolinska Institutet.
Uppsala universitet.
KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.
KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

The role of calcium signaling for development of early vertebrates is well documented, but little is known about its role in mammalian embryogenesis. We have used explanted embryonic rat kidneys to study the role of calcium for branching morphogenesis, a process that depends on reciprocal interaction between mesenchymal and epithelial ureteric bud cells. We recorded a spontaneous calcium activity characterized by stochastic and irregular calcium spikes, in the mesenchymal cells. This activity is due to calcium release from the endoplasmic reticulum (ER). Depletion of ER calcium stores results in down-regulation of the calcium activity, retardation of branching morphogenesis and formation of primitive nephrons, but has no effect on cell proliferation. We propose that the excretion of morphogenic factors that mediate the interaction between 26 the mesenchymal and epithelial cells, which initiate branching morphogenesis, is calcium dependent. In support of this we demonstrate expression of the calcium dependent excretory protein synaptotagmin1.

Keywords [en]
Calcium signaling, kidney development, embryonic kidney, synaptotagmin
National Category
Developmental Biology
Identifiers
URN: urn:nbn:se:kth:diva-207581OAI: oai:DiVA.org:kth-207581DiVA, id: diva2:1097116
Funder
Swedish Research Council, 2013-6041Swedish Research Council, 2015-04198
Note

QC 20170523

Available from: 2017-05-22 Created: 2017-05-22 Last updated: 2017-05-23Bibliographically approved
In thesis
1. High-resolution imaging of kidney tissue samples
Open this publication in new window or tab >>High-resolution imaging of kidney tissue samples
2017 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

The kidney is one of the most important and complex organs in the human body, filtering hundreds of litres of blood daily. Kidney disease is one of the fastest growing causes of death in the modern world, and this motivates extensive research for better understanding the function of the kidney in health and disease. Some of the most important cellular structures for blood filtration in the kidney are of very small dimensions (on the sub-200 nm scale), and thus electron microscopy has been the only method of choice to visualize these minute structures. In one study, we show for the first time that by combining optical clearing with STED microscopy, protein localizations in the slit diaphragm of the kidney, a structure around 75 nanometers in width, can now be resolved using light microscopy. In a second study, a novel sample preparation method, expansion microscopy, is utilized to physically expand kidney tissue samples. Expansion improves the effective resolution by a factor of 5, making it possible to resolve podocyte foot processes and the slit diaphragm using confocal microscopy. We also show that by combining expansion microscopy and STED microscopy, the effective resolution can be improved further. In a third study, influences on the development of the kidney were studied. There is substantial knowledge regarding what genes (growth factors, receptors etc.) are important for the normal morphogenesis of the kidney. Less is known regarding the physiology behind how paracrine factors are secreted and delivered in the developing kidney. By depleting calcium transients in explanted rat kidneys, we show that calcium is important for the branching morphogenesis of the ureteric tree. Further, the study shows that the calcium-dependent initiator of exocytosis, synaptotagmin, is expressed in the metanephric mesenchyme of the developing kidney, indicating that it could have a role in the secretion of paracrine growth factors, such as GDNF, to drive the branching.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2017. p. 34
Series
TRITA-FYS, ISSN 0280-316X ; 2017:28
Keywords
Super Resolution Microscopy, Kidney, Imaging, Fluorescence, Kidney development, Calcium
National Category
Biophysics
Research subject
Biological Physics
Identifiers
urn:nbn:se:kth:diva-207577 (URN)978-91-7729-456-6 (ISBN)
Presentation
2017-06-15, Air-Fire, Science for Life Laboratories, Tomtebodavägen 23A, Solna, 09:30 (English)
Opponent
Supervisors
Funder
Swedish Foundation for Strategic Research , RIF14-0091Swedish Research Council, 2013-6041
Note

QC 20170523

Available from: 2017-05-23 Created: 2017-05-22 Last updated: 2017-05-23Bibliographically approved

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