scholarly journals Mapping out Min protein patterns in fully confined fluidic chambers

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Yaron Caspi ◽  
Cees Dekker

The bacterial Min protein system provides a major model system for studying reaction-diffusion processes in biology. Here we present the first in vitro study of the Min system in fully confined three-dimensional chambers that are lithography-defined, lipid-bilayer coated and isolated through pressure valves. We identify three typical dynamical behaviors that occur dependent on the geometrical chamber parameters: pole-to-pole oscillations, spiral rotations, and traveling waves. We establish the geometrical selection rules and show that, surprisingly, Min-protein spiral rotations govern the larger part of the geometrical phase diagram. Confinement as well as an elevated temperature reduce the characteristic wavelength of the Min patterns, although even for confined chambers with a bacterial-level viscosity, the patterns retain a ~5 times larger wavelength than in vivo. Our results provide an essential experimental base for modeling of intracellular Min gradients in bacterial cell division as well as, more generally, for understanding pattern formation in reaction-diffusion systems.

2012 ◽  
Vol 109 (38) ◽  
pp. 15283-15288 ◽  
Author(s):  
Jakob Schweizer ◽  
Martin Loose ◽  
Mike Bonny ◽  
Karsten Kruse ◽  
Ingolf Mönch ◽  
...  

In the living cell, proteins are able to organize space much larger than their dimensions. In return, changes of intracellular space can influence biochemical reactions, allowing cells to sense their size and shape. Despite the possibility to reconstitute protein self-organization with only a few purified components, we still lack knowledge of how geometrical boundaries affect spatiotemporal protein patterns. Following a minimal systems approach, we used purified proteins and photolithographically patterned membranes to study the influence of spatial confinement on the self-organization of the Min system, a spatial regulator of bacterial cytokinesis, in vitro. We found that the emerging protein pattern responds even to the lateral, two-dimensional geometry of the membrane such that, as in the three-dimensional cell, Min protein waves travel along the longest axis of the membrane patch. This shows that for spatial sensing the Min system does not need to be enclosed in a three-dimensional compartment. Using a computational model we quantitatively analyzed our experimental findings and identified persistent binding of MinE to the membrane as requirement for the Min system to sense geometry. Our results give insight into the interplay between geometrical confinement and biochemical patterns emerging from a nonlinear reaction–diffusion system.


eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Piotr Szwedziak ◽  
Qing Wang ◽  
Tanmay A M Bharat ◽  
Matthew Tsim ◽  
Jan Löwe

Membrane constriction is a prerequisite for cell division. The most common membrane constriction system in prokaryotes is based on the tubulin homologue FtsZ, whose filaments in E. coli are anchored to the membrane by FtsA and enable the formation of the Z-ring and divisome. The precise architecture of the FtsZ ring has remained enigmatic. In this study, we report three-dimensional arrangements of FtsZ and FtsA filaments in C. crescentus and E. coli cells and inside constricting liposomes by means of electron cryomicroscopy and cryotomography. In vivo and in vitro, the Z-ring is composed of a small, single-layered band of filaments parallel to the membrane, creating a continuous ring through lateral filament contacts. Visualisation of the in vitro reconstituted constrictions as well as a complete tracing of the helical paths of the filaments with a molecular model favour a mechanism of FtsZ-based membrane constriction that is likely to be accompanied by filament sliding.


Materials ◽  
2018 ◽  
Vol 11 (10) ◽  
pp. 2047 ◽  
Author(s):  
Yunpeng Bai ◽  
Takahiro Kanno ◽  
Hiroto Tatsumi ◽  
Kenichi Miyamoto ◽  
Jingjing Sha ◽  
...  

This study evaluated the feasibility of a novel three-dimensional (3D) porous composite of uncalcined and unsintered hydroxyapatite (u-HA) and poly-d/l-lactide (PDLLA) (3D-HA/PDLLA) for the bony regenerative biomaterial in maxillofacial surgery, focusing on cellular activities and osteoconductivity properties in vitro and in vivo. In the in vitro study, we assessed the proliferation and ingrowth of preosteoblastic cells (MC3T3-E1 cells) in 3D-HA/PDLLA biomaterials using 3D cell culture, and the results indicated enhanced bioactive proliferation. After osteogenic differentiation of those cells on 3D-HA/PDLLA, the osteogenesis marker genes runt-related transcription factor-2 (Runx2), and Sp7 (Osterix) were upregulated. For the in vivo study, we evaluated the utility of 3D-HA/PDLLA biomaterials compared to the conventional bone substitute of beta-tricalcium phosphate (β-TCP) in rats with critical mandibular bony defects. The implantation of 3D-HA/PDLLA biomaterials resulted in enhanced bone regeneration, by inducing high osteoconductivity as well as higher β-TCP levels. Our study thus showed that the novel composite, 3D-HA/PDLLA, is an excellent bioactive/bioresorbable biomaterial for use as a cellular scaffold, both in vitro and in vivo, and has utility in bone regenerative therapy, such as for patients with irregular maxillofacial bone defects.


1999 ◽  
Vol 02 (03) ◽  
pp. 221-276 ◽  
Author(s):  
James Tabony ◽  
Laurent Vuillard ◽  
Cyril Papaseit

Chemically dissipative or Turing processes, have been predicted by theoreticians as a way by which an initially homogenous solution of chemicals or biochemicals can spontaneously self-organise and give rise to a macroscopic pattern by way of a combination of reaction and diffusion. They have been advanced as a possible underlying process for biological self-organisation and pattern formation. Until now, there have been no examples of in vitro biological substances showing this type of behaviour. Evidence is presented that microtubule solutions in vitro self-organise in this manner and that similar processes may occur in vivo during embryogenesis.


2019 ◽  
Vol 877 ◽  
pp. 759-823 ◽  
Author(s):  
Robert A. Van Gorder ◽  
Hyunyeon Kim ◽  
Andrew L. Krause

We study spatial and spatio-temporal pattern formation emergent from reaction–diffusion–advection systems formed by considering reaction–diffusion systems coupled to prescribed fluid flows. While there have been a number of studies on the planar dynamics of such systems and the resulting instabilities and spatio-temporal patterning in the plane, less has been done on complicated flows in complex domains. We consider a general approach for the study of bounded domains in order to model two- and three-dimensional geometries which are more likely to be of relevance for modelling dynamics within fluid vessels used in experiments. Considering a variety of problem geometries with finite cross-sections, such as two-dimensional channels, three-dimensional ducts and three-dimensional pipes, we demonstrate the role cross-section geometry plays in pattern formation under such systems. We find that the generic instability is that of an oscillatory or wave Turing instability, resulting in patterns which change in time, often being advected with the fluid flow. As in previous works, we observe a change in patterns formed when progressing from zero to weak to strong advection for uniform advection across the domain, with particularly strong advection destroying patterns. One novel finding is that heterogeneous fluid flow can induce qualitatively different patterns across the domain. For instance, Poiseuille flow with maximal advection in the centre of a vessel and zero advection at the boundary of a vessel is shown to exhibit patterns in the centre of the vessel which are different from patterns near the boundary, with differences attributed to the differential local advection within each region of the vessel. Additionally, we observe sheared patterns, which appear due to gradients in the fluid velocity, and cannot be obtained via any kind of uniform flow. Finally we also explore flow in more complex domains, including wavy-walled channels, continuous stirred-tank reactors, U-shaped pipes and a toroidal domain, in order to demonstrate behaviours when the flow is both heterogeneous and bidirectional, as well as to demonstrate that our results still apply for complex finite domains. Our analysis suggests that such non-trivial advection results in moving patterns which are more complex than observed in simpler reaction–diffusion–advection, and may be more characteristic of realistic flow regimes in biological media.


2017 ◽  
Vol 32 (3) ◽  
pp. 342-348
Author(s):  
Weiwei Jin ◽  
Yanxue Yin ◽  
Bo Zhang ◽  
Heng Mei ◽  
Huafang Wang ◽  
...  

Tissue factor is an upstream component of the cascade and a high-expressing factor under phathological conditions. In this study, a tissue factor cascade-targeted strategy for inducing local thrombosis was developed by combining ENP-HMME and photochemistry. In vitro study showed that protein EGFP-EGF1 conjugation to the nanoparticles could significantly contribute to the uptake of nanoparticles by tissue factor over-expressed brain capillary endothelial cells. Three-dimensional imaging and specklegram of brains in vivo showed that tissue factor cascade-targeted strategy successfully induced thrombosis of expected position. As shown by the in vivo multispectral fluorescent imaging, when ENP-HMME was combined with photochemistry, higher accumulation in the infarction hemisphere was observed, which might suggest that the photochemistry inducing tissue factor cascade recruited more ENP-HMME than HMME-loaded nanoparticles (NP-HMME). The data indicated the tissue factor cascade-targeted strategy has potential to induce local thrombosis, and might be applied in the treatment of a variety of hypervascular diseases.


Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.


2014 ◽  
Author(s):  
Ivo Dumic-Cule ◽  
Dunja Rogic ◽  
Damir Jezek ◽  
Lovorka Grgurevic ◽  
Slobodan Vukicevic

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