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Three-Dimensional Visualization of the Podocyte Actin Network Using Integrated Membrane Extraction, Electron

Chengqing Qu¹, Robyn Roth², Pongpratch Puapatanakul³

¹Department of Mechanical Engineering, National Science Foundation Science and Technology Center for Engineering Mechanobiology, Washington University in St. Louis, St. Louis, Missouri.

Journal of the American Society of Nephrology : JASN

|November 11, 2021

Summary

Researchers developed a new high-resolution imaging technique to visualize the 3D actin cytoskeleton in mouse podocytes, revealing novel structural details of actin networks in vivo.

Keywords:

actin cytoskeleton intermediate filaments podocyte

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Area of Science:

Cell Biology
Biophysics
Nephrology

Background:

Actin stress fibers are crucial for cell shape and adhesion in podocytes, but their in vivo structure and function remain poorly understood.
Changes in actin stress fibers are implicated in podocyte injury and disease, yet in vivo visualization has been challenging.
A high-resolution imaging technology is needed to study the in vivo role of actin stress fibers in podocytes.

Purpose of the Study:

To develop and apply a novel technique for high-resolution 3D imaging of the actin cytoskeleton in mouse podocytes.
To elucidate the detailed structure of actin networks within podocyte foot processes and cell bodies in vivo.
To investigate the role of podocyte actin structures in maintaining glomerular filtration barrier integrity.

Main Methods:

Developed a novel visualization technique integrating membrane extraction, focused ion-beam scanning electron microscopy, and machine learning image segmentation.
Applied the technique to image the 3D cytoskeletal network in mouse podocytes.
Identified proteins comprising the cytoskeletal network.

Main Results:

Achieved the first detailed 3D visualization of the mouse podocyte actin cytoskeleton in vivo.
Observed actin cables and intermediate filaments connecting foot processes to contractile structures at the cell body periphery.
Discovered that actin cables form a continuous, mesh-like sheet within foot processes, incorporating slit diaphragms.

Conclusions:

The novel technique provides unprecedented insight into the 3D organization of podocyte actin networks.
Findings support the gel compression hypothesis regarding foot process function in counterbalancing hydrodynamic forces.
The study offers new understanding of how podocytes sense and respond to mechanical cues.