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Published on: March 27, 2017
Observation of angiotensin II-induced changes in fixed and live mesangial cells by atomic force microscopy
Gi-Ja Lee1, Eun-Jin Park, Samjin Choi
1Department of Biomedical Engineering, College of Medicine, Kyung Hee University, Dongdaemun-gu, Seoul 130-701, Republic of Korea.
Abstract:
Glomerular mesangial cells (MCs) are centrally located in the glomerulus. MCs control not only glomerular filtration, but also the response to local injury, including cell proliferation and basement membrane remodeling. Angiotensin II (Ang II) plays an important role in kidney function regulation, and participates in the progression of renal damage, as well as mesangial injury. However, studies on Ang II effects on MCs have used indirect methods, such as gene and protein expression after MC injury. In this study, we visually observed structural and mechanical changes to MC after Ang II treatment using atomic force microscopy (AFM). We obtained AFM topography and deflection images of live MCs, as well as fixed MCs in liquid, before and after Ang II treatment. Real-time imaging showed the dynamic movement of live MCs induced by Ang II. Changes in MC elastic property after Ang II treatment were measured using force-distance curves. AFM images of fixed and live MCs showed that cells contracted after Ang II exposure, with the nucleus height increasing within 20 min of Ang II stimulation. Force-distance analysis showed that Ang II caused MCs to stiffen (p < 0.0001). In conclusion, we demonstrated that AFM is an effective tool for real-time monitoring of live cell responses to drugs and stimuli.
Insights
Atomic force microscopy visually tracked live glomerular mesangial cells (MCs) responding to Angiotensin II (Ang II). This study revealed Ang II causes MCs to contract and stiffen, offering new insights into kidney injury mechanisms.
Area of Science:
- Cell biology
- Nephrology
- Biophysics
Background:
- Glomerular mesangial cells (MCs) are crucial for kidney filtration and injury response.
- Angiotensin II (Ang II) significantly impacts kidney function and mesangial injury.
- Previous studies on Ang II effects on MCs relied on indirect methods.
Purpose of the Study:
- To visually observe structural and mechanical changes in MCs after Ang II treatment using AFM.
- To investigate real-time cellular responses to Ang II stimulation.
- To assess the impact of Ang II on MC mechanical properties.
Main Methods:
- Utilized Atomic Force Microscopy (AFM) for topography and deflection imaging of live and fixed MCs.
- Performed real-time imaging to capture dynamic MC movement.
- Applied force-distance curves to measure changes in MC elastic properties.
Main Results:
- Ang II treatment induced visible contraction in MCs.
- Nucleus height increased within 20 minutes of Ang II stimulation.
- Force-distance analysis demonstrated significant stiffening of MCs post-Ang II exposure (p < 0.0001).
Conclusions:
- AFM provides an effective platform for real-time monitoring of live cell responses to stimuli.
- Ang II directly alters MC structure and mechanical properties, contributing to kidney injury.
- This study offers a novel visual and mechanical approach to understanding drug-induced cellular changes in the kidney.
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