Observation of angiotensin II-induced changes in fixed and live mesangial cells by atomic force microscopy.
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.
Lee
Gi-Ja
GJ
Department of Biomedical Engineering, College of Medicine, Kyung Hee University, Dongdaemun-gu, Seoul 130-701, Republic of Korea.
Park
Eun-Jin
EJ
Choi
Samjin
S
Park
Jeong-Hoon
JH
Jeong
Kyung-Hwan
KH
Kim
Kyung-Sook
KS
Lee
Sang-Ho
SH
Park
Hun-Kuk
HK
D000804:Angiotensin II / Q000378:metabolism*; D006801:Humans; D050527:Mesangial Cells / Q000187:drug effects* / Q000648:ultrastructure; D018625:Microscopy, Atomic Force / Q000379:methods
D000804:Angiotensin II