Cross-bridge cycling gives rise to spatiotemporal heterogeneity of dynamic subcellular mechanics in cardiac myocytes probed with atomic force microscopy

2010 ◽  
Vol 298 (3) ◽  
pp. H853-H860 ◽  
Author(s):  
Evren U. Azeloglu ◽  
Kevin D. Costa

To study how the dynamic subcellular mechanical properties of the heart relate to the fundamental underlying process of actin-myosin cross-bridge cycling, we developed a novel atomic force microscope elastography technique for mapping spatiotemporal stiffness of isolated, spontaneously beating neonatal rat cardiomyocytes. Cells were indented repeatedly at a rate close but unequal to their contractile frequency. The resultant changes in pointwise apparent elastic modulus cycled at a predictable envelope frequency between a systolic value of 26.2 ± 5.1 kPa and a diastolic value of 7.8 ± 4.1 kPa at a representative depth of 400 nm. In cells probed along their major axis, spatiotemporal changes in systolic stiffness displayed a heterogeneous pattern, reflecting the banded sarcomeric structure of underlying myofibrils. Treatment with blebbistatin eliminated contractile activity and resulted in a uniform apparent modulus of 6.5 ± 4.8 kPa. This study represents the first quantitative dynamic mechanical mapping of beating cardiomyocytes. The technique provides a means of probing the micromechanical effects of disease processes and pharmacological treatments on beating cardiomyocytes, providing new insights and relating subcellular cardiac structure and function.

2016 ◽  
Vol 3 ◽  
pp. 184954351667534 ◽  
Author(s):  
Neerajha Nagarajan ◽  
Varun Vyas ◽  
Bryan D Huey ◽  
Pinar Zorlutuna

The ability to modulate cardiomyocyte contractility is important for bioengineering applications ranging from heart disease treatments to biorobotics. In this study, we examined the changes in contraction frequency of neonatal rat cardiomyocytes upon single-cell-level nanoscale mechanical stimulation using atomic force microscopy. To measure the response of same density of cells, they were micropatterned into micropatches of fixed geometry. To examine the effect of the substrate stiffness on the behavior of cells, they were cultured on a stiffer and a softer surface, glass and poly (dimethylsiloxane), respectively. Upon periodic cyclic stimulation of 300 nN at 5 Hz, a significant reduction in the rate of synchronous contraction of the cell patches on poly(dimethylsiloxane) substrates was observed with respect to their spontaneous beat rate, while the cell patches on glass substrates maintained or increased their contraction rate after the stimulation. On the other hand, single cells mostly maintained their contraction rate and could only withstand a lower magnitude of forces compared to micropatterned cell patches. This study reveals that the contraction behavior of cardiomyocytes can be modulated mechanically through cyclic nanomechanical stimulation, and the degree and mode of this modulation depend on the cell connectivity and substrate mechanical properties.


2020 ◽  
Vol 92 (6) ◽  
pp. 977-984
Author(s):  
Mayya V. Kulikova ◽  
Albert B. Kulikov ◽  
Alexey E. Kuz’min ◽  
Anton L. Maximov

AbstractFor previously studied Fischer–Tropsch nanosized Fe catalyst slurries, polymer compounds with or without polyconjugating structures are used as precursors to form the catalyst nanomatrix in situ, and several catalytic experiments and X-ray diffraction and atomic force microscopy measurements are performed. The important and different roles of the paraffin molecules in the slurry medium in the formation and function of composite catalysts with the two types of aforementioned polymer matrices are revealed. In the case of the polyconjugated polymers, the alkanes in the medium are “weakly” coordinated with the metal-polymer composites, which does not affect the effectiveness of the polyconjugated polymers. Otherwise, alkane molecules form a “tight” surface layer around the composite particles, which create transport complications for the reagents and products of Fischer-Tropsch synthesis and, in some cases, can change the course of the in situ catalyst formation.


2018 ◽  
Author(s):  
Yuchen Long ◽  
Ibrahim Cheddadi ◽  
Vincent Mirabet ◽  
Gabriella Mosca ◽  
Mathilde Dumond ◽  
...  

Cell-to-cell heterogeneity prevails in many biological systems, although its origin and function are often unclear. Cell hydrostatic pressure, alias turgor pressure, is essential in physiology and morphogenesis, and its spatial variations are often overlooked. Here, based on a mathematical model describing cell mechanics and water movement in a plant tissue, we predict that cell pressure anticorrelates with cell neighbour number. Using atomic force microscopy, we confirm this prediction in the Arabidopsis shoot apical meristem, a population of stem cells that generate all plant aerial organs. Pressure is predicted to correlate either positively or negatively with cellular growth rate depending on osmotic drive, cell wall extensibility, and hydraulic conductivity. The meristem exhibits one of these two regimes depending on conditions, suggesting that, in this tissue, water conductivity is non-negligible in growth control. Our results illustrate links between local topology, cell mechanical state and cell growth, with potential roles in tissue homeostasis.


1994 ◽  
pp. 264-275
Author(s):  
Kunio Takeyasu ◽  
Jose K. Paul ◽  
Mehdi Ganjeizadeh ◽  
M. Victor Lemas ◽  
Shusheng Wang ◽  
...  

2001 ◽  
Vol 7 (S2) ◽  
pp. 124-125
Author(s):  
Christopher A. Siedlecki

A widely accepted tenet of biomaterials research is that the initial step following contact of a synthetic material with blood is the rapid adsorption of plasma proteins. The composition of this adsorbed protein layer is dependent on a variety of factors, including the surface properties of the implant material and the nature of the adsorbing proteins, and the composition and function of this protein layer is important in the subsequent biological response and ultimately the success or failure of the implanted material. While a great amount of effort has gone into developing structure/function responses for implanted biomaterials, there is still much about the molecular level interactions to be determined. We utilized atomic force microscopy (AFM) to investigate the molecular-level interactions of proteins with model biomaterial substrates. The AFM is unique in that it offers the opportunity to characterize interfacial environments, determine material properties, measure protein/surface interaction forces, and visualize the tertiary structure of adsorbed proteins.


2020 ◽  
Vol 202 (12) ◽  
Author(s):  
Albertus Viljoen ◽  
Johann Mignolet ◽  
Felipe Viela ◽  
Marion Mathelié-Guinlet ◽  
Yves F. Dufrêne

ABSTRACT Microbial adhesion and biofilm formation are usually studied using molecular and cellular biology assays, optical and electron microscopy, or laminar flow chamber experiments. Today, atomic force microscopy (AFM) represents a valuable addition to these approaches, enabling the measurement of forces involved in microbial adhesion at the single-molecule level. In this minireview, we discuss recent discoveries made applying state-of-the-art AFM techniques to microbial specimens in order to understand the strength and dynamics of adhesive interactions. These studies shed new light on the molecular mechanisms of adhesion and demonstrate an intimate relationship between force and function in microbial adhesins.


Author(s):  
Liang Ma ◽  
Meixiang Xu ◽  
Andres F. Oberhauser

The activity of proteins and their complexes often involves the conversion of chemical energy (stored or supplied) into mechanical work through conformational changes. Mechanical forces are also crucial for the regulation of the structure and function of cells and tissues. Thus, the shape of eukaryotic cells is the result of cycles of mechano-sensing, mechano-transduction, and mechano-response. Recently developed single-molecule atomic force microscopy (AFM) techniques can be used to manipulate single molecules, both in real time and under physiological conditions, and are ideally suited to directly quantify the forces involved in both intra- and intermolecular protein interactions. In combination with molecular biology and computer simulations, these techniques have been applied to characterize the unfolding and refolding reactions in a variety of proteins, such as titin (an elastic mechano-sensing protein found in muscle) and polycystin-1 (PC1, a mechanosensor found in the kidney).


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