3,218 research outputs found
Domain Growth Kinetics in a Cell-sized Liposome
We investigated the kinetics of domain growth on liposomes consisting of a
ternary mixture (unsaturated phospholipid, saturated phospholipid, and
cholesterol) by temperature jump. The domain growth process was monitored by
fluorescence microscopy, where the growth was mediated by the fusion of domains
through the collision. It was found that an average domain size r develops with
time t as r ~ t^0.15, indicating that the power is around a half of the
theoretical expectation deduced from a model of Brownian motion on a
2-dimensional membrane. We discuss the mechanism of the experimental scaling
behavior by considering the elasticity of the membrane
Line tensions, correlation lengths, and critical exponents in lipid membranes near critical points
Membranes containing a wide variety of ternary mixtures of high chain-melting
temperature lipids, low chain-melting temperature lipids, and cholesterol
undergo lateral phase separartion into coexisting liquid phases at a
miscibility transition. When membranes are prepared from a ternary lipid
mixture at a critical composition, they pass through a miscibility critical
point at the transition temperature. Since the critical temperature is
typically on the order of room temperature, membranes provide an unusual
opportunity in which to perform a quantitative study of biophysical systems
that exhibit critical phenomena in the two-dimensional Ising universality
class. As a critical point is approached from either high or low temperature,
the scale of fluctuations in lipid composition, set by the correlation length,
diverges. In addition, as a critical point is approached from low temperature,
the line tension between coexisting phases decreases to zero. Here we
quantitatively evaluate the temperature dependence of line tension between
liquid domains and of fluctuation correlation lengths in lipid membranes in
order to extract a critical exponent, nu. We obtain nu=1.2 plus or minus 0.2,
consistent with the Ising model prediction nu=1. We also evaluate the
probability distributions of pixel intensities in fluoresence images of
membranes. From the temperature dependence of these distributions above the
critical temperature, we extract an independent critical exponent beta=0.124
plus or minus 0.03 which is consistent with the Ising prediction of beta=1/8.Comment: 22 pages, 7 figure
Mitochondrial dysfunction leads to nuclear genome instability: A link through iron-sulfur clusters
Mutations and deletions in the mitochondrial genome (mtDNA), as well as
instability of the nuclear genome, are involved in multiple human diseases. Here we
report that in Saccharomyces cerevisiae, loss of mtDNA leads to nuclear genome
instability, through a process of cell cycle arrest and selection we define as a cellular
crisis. This crisis is not mediated by the absence of respiration, but instead correlates with
a reduction in the mitochondrial membrane potential. Analysis of cells undergoing this
crisis identified a defect in iron-sulfur cluster (ISC) biogenesis, which requires normal
mitochondrial function. We found that down-regulation of non-mitochondrial ISC protein
biogenesis was sufficient to cause increased genomic instability in cells with intact
mitochondrial function. These results suggest mitochondrial dysfunction stimulates
nuclear genome instability by inhibiting the production of ISC-containing protein(s),
which are required for maintenance of nuclear genome integrity
Lateral phase separation in mixtures of lipids and cholesterol
In an effort to understand "rafts" in biological membranes, we propose phenomenological models for saturated and unsaturated lipid mixtures, and lipid-cholesterol mixtures. We consider simple couplings between the local composition and internal membrane structure, and their influence on transitions between liquid and gel membrane phases. Assuming that the gel transition temperature of the saturated lipid is shifted by the presence of the unsaturated lipid, and that cholesterol acts as an external field on the chain melting transition, a variety of phase diagrams are obtained. The phase diagrams for binary mixtures of saturated/unsaturated lipids and lipid/cholesterol are in semi-quantitative agreement with the experiments. Our results also apply to regions in the ternary phase diagram of lipid/lipid/cholesterol systems
Stable patterns of membrane domains at corrugated substrates
Multi-component membranes such as ternary mixtures of lipids and cholesterol
can exhibit coexistence regions between two liquid phases. When such membranes
adhere to a corrugated substrate, the phase separation process strongly depends
on the interplay between substrate topography, bending rigidities, and line
tension of the membrane domains as we show theoretically via energy
minimization and Monte Carlo simulations. For sufficiently large bending
rigidity contrast between the two membrane phases, the corrugated substrate
truncates the phase separation process and leads to a stable pattern of
membrane domains. Our theory is consistent with recent experimental
observations and provides a possible control mechanism for domain patterns in
biological membranes.Comment: to appear in Physical Review Letter
Correlation functions quantify super-resolution images and estimate apparent clustering due to over-counting
We present an analytical method to quantify clustering in super-resolution
localization images of static surfaces in two dimensions. The method also
describes how over-counting of labeled molecules contributes to apparent
self-clustering and how the effective lateral resolution of an image can be
determined. This treatment applies to clustering of proteins and lipids in
membranes, where there is significant interest in using super-resolution
localization techniques to probe membrane heterogeneity. When images are
quantified using pair correlation functions, the magnitude of apparent
clustering due to over-counting will vary inversely with the surface density of
labeled molecules and does not depend on the number of times an average
molecule is counted. Over-counting does not yield apparent co-clustering in
double label experiments when pair cross-correlation functions are measured. We
apply our analytical method to quantify the distribution of the IgE receptor
(Fc{\epsilon}RI) on the plasma membranes of chemically fixed RBL-2H3 mast cells
from images acquired using stochastic optical reconstruction microscopy (STORM)
and scanning electron microscopy (SEM). We find that apparent clustering of
labeled IgE bound to Fc{\epsilon}RI detected with both methods arises from
over-counting of individual complexes. Thus our results indicate that these
receptors are randomly distributed within the resolution and sensitivity limits
of these experiments.Comment: 22 pages, 5 figure
Molecular rheometry: direct determination of viscosity in L-o and L-d lipid phases via fluorescence lifetime imaging
Understanding of cellular regulatory pathways that involve lipid membranes requires the detailed knowledge of their physical state and structure. However, mapping the viscosity and diffusion in the membranes of complex composition is currently a non-trivial technical challenge. We report fluorescence lifetime spectroscopy and imaging (FLIM) of a meso-substituted BODIPY molecular rotor localised in the leaflet of model membranes of various lipid compositions. We prepare large and giant unilamellar vesicles (LUVs and GUVs) containing phosphatidylcholine (PC) lipids and demonstrate that recording the fluorescence lifetime of the rotor allows us to directly detect the viscosity of the membrane leaflet and to monitor the influence of cholesterol on membrane viscosity in binary and ternary lipid mixtures. In phase-separated 1,2-dioleoyl-sn-glycero-3-phosphocholine-cholesterol–sphingomyelin GUVs we visualise individual liquid ordered (Lo) and liquid disordered (Ld) domains using FLIM and assign specific microscopic viscosities to each domain. Our study showcases the power of FLIM with molecular rotors to image microviscosity of heterogeneous microenvironments in complex biological systems, including membrane-localised lipid rafts
Molecular motors robustly drive active gels to a critically connected state
Living systems often exhibit internal driving: active, molecular processes
drive nonequilibrium phenomena such as metabolism or migration. Active gels
constitute a fascinating class of internally driven matter, where molecular
motors exert localized stresses inside polymer networks. There is evidence that
network crosslinking is required to allow motors to induce macroscopic
contraction. Yet a quantitative understanding of how network connectivity
enables contraction is lacking. Here we show experimentally that myosin motors
contract crosslinked actin polymer networks to clusters with a scale-free size
distribution. This critical behavior occurs over an unexpectedly broad range of
crosslink concentrations. To understand this robustness, we develop a
quantitative model of contractile networks that takes into account network
restructuring: motors reduce connectivity by forcing crosslinks to unbind.
Paradoxically, to coordinate global contractions, motor activity should be low.
Otherwise, motors drive initially well-connected networks to a critical state
where ruptures form across the entire network.Comment: Main text: 21 pages, 5 figures. Supplementary Information: 13 pages,
8 figure
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