Abstract: SA-PO194

Acute Kidney Slices as a Tool to Dissect Calcium Signals in Podocytes

Session Information

  • Glomerular: Cell Biology
    November 04, 2017 | Location: Hall H, Morial Convention Center
    Abstract Time: 10:00 AM - 10:00 AM

Category: Glomerular

  • 1003 Glomerular: Cell Biology

Authors

  • Binz, Julia, University Hospital Cologne, Cologne, Germany
  • Khalili, Hadiseh, University Hospital of Cologne, Köln, Germany
  • Schermer, Bernhard, University Hospital Cologne, Cologne, Germany
  • Benzing, Thomas, University of Cologne, Köln, Germany
  • Hackl, Matthias, University Hospital Cologne, Cologne, Germany
Background

Calcium signaling in the kidney has been a focus of research for many years. Especially calcium handling in podocytes has been of great interest since it is implicated to play a role in podocyte injury. As isolated podocytes in cell culture are lacking their physiologic microenviroment and isolated glomeruli are prepared with several treatment steps, acute kidney slices (AKS) represent an ex vivo model where the structural complexity of the glomerulus is preserved and which can be prepared within minutes after sacrificing the animal.

Methods

Prior to the preparation of AKS 8-week old C57BL6 mice expressing GCaMP3 under the Pod:Cre promoter were treated with Adriamycin (25mg/kg) for five days. Untreated mice were used as control group. For AKS preparation mice were sacrificed, the kidneys were removed and cut into 500 µm thick slices. During imaging a multiphoton laser was used at 940nm and AKS were treated with an angiotensin-II containing solution to induce calcium signaling in podocytes. As control experiments we used laser-induced calcium signaling and propidium iodide to show that podocytes remain viable and able to produce calcium signals.

Results

The podocytes with genetically encoded calcium indicator are viable and able to produce a calcium wave after laser injury as published in vivo. High doses of angiotensin-II elicit calcium signals in podocytes of untreated animals. Furthermore, there is an increase in calcium response after induction of Adriamycin nephropathy.

Conclusion

Acute slices of the kidney are a great tool to study podocytes, retaining the complex microarchitecture of the glomerulus, and provide results comparable to in vivo imaging. They allow testing of different dosages and compounds on individual kidney slices as the two kidneys provide enough material for at least 5 slices each. The technical setup needed for preparation is limited and the technique can be quickly taught. A confocal imaging setup is sufficient to image AKS. Therefore, AKS have the potential to become a widely used tool in glomerular research.