24 research outputs found
Cdc42 controls the dilation of the exocytotic fusion pore by regulating membrane tension.
Membrane fusion underlies multiple processes, including exocytosis of hormones and neurotransmitters. Membrane fusion starts with the formation of a narrow fusion pore. Radial expansion of this pore completes the process and allows fast release of secretory compounds, but this step remains poorly understood. Here we show that inhibiting the expression of the small GTPase Cdc42 or preventing its activation with a dominant negative Cdc42 construct in human neuroendocrine cells impaired the release process by compromising fusion pore enlargement. Consequently the mode of vesicle exocytosis was shifted from full-collapse fusion to kiss-and-run. Remarkably, Cdc42-knockdown cells showed reduced membrane tension, and the artificial increase of membrane tension restored fusion pore enlargement. Moreover, inhibiting the motor protein myosin II by blebbistatin decreased membrane tension, as well as fusion pore dilation. We conclude that membrane tension is the driving force for fusion pore dilation and that Cdc42 is a key regulator of this force.journal articleresearch support, non-u.s. gov't2014 Oct 152014 08 20importe
Rate-dependent Ca2+ signalling underlying the force-frequency response in rat ventricular myocytes: A coupled electromechanical modeling study
Rate-dependent effects on the Ca2+ sub-system in a rat ventricular myocyte are investigated. Here,
we employ a deterministic mathematical model describing various Ca2+ signalling pathways under
voltage clamp (VC) conditions, to better understand the important role of calmodulin (CaM) in modulating
the key control variables Ca2+/calmodulin-dependent protein kinase-II (CaMKII), calcineurin
(CaN), and cyclic adenosine monophosphate (cAMP) as they affect various intracellular targets. In
particular, we study the frequency dependence of the peak force generated by the myofilaments, the
force-frequency response (FFR). Our cell model incorporates frequency-dependent CaM-mediated spatially heterogenous interaction
of CaMKII and CaN with their principal targets (dihydropyridine (DHPR) and ryanodine (RyR) receptors
and the SERCA pump). It also accounts for the rate-dependent effects of phospholamban
(PLB) on the SERCA pump; the rate-dependent role of cAMP in up-regulation of the L-type Ca2+
channel (ICa;L); and the enhancement in SERCA pump activity via phosphorylation of PLB.Our model reproduces positive peak FFR observed in rat ventricular myocytes during voltage-clamp
studies both in the presence/absence of cAMP mediated -adrenergic stimulation. This study provides
quantitative insight into the rate-dependence of Ca2+-induced Ca2+-release (CICR) by investigating
the frequency-dependence of the trigger current (ICa;L) and RyR-release. It also highlights the relative
role of the sodium-calcium exchanger (NCX) and the SERCA pump at higher frequencies, as well
as the rate-dependence of sarcoplasmic reticulum (SR) Ca2+ content. A rigorous Ca2+ balance
imposed on our investigation of these Ca2+ signalling pathways clarifies their individual roles. Here,
we present a coupled electromechanical study emphasizing the rate-dependence of isometric force
developed and also investigate the temperature-dependence of FFR. Our model provides mechanistic biophysically based explanations for the rate-dependence of CICR,
generating useful and testable hypotheses. Although rat ventricular myocytes exhibit a positive peak
FFR in the presence/absence of beta-adrenergic stimulation, they show a characteristic increase in the
positive slope in FFR due to the presence of Norepinephrine or Isoproterenol. Our study identifies
cAMP-mediated stimulation, and rate-dependent CaMKII-mediated up-regulation of ICa;L as the key
mechanisms underlying the aforementioned positive FFR
Local contributions to the Euler-Poincare characteristic of a set
International audienceThe Euler–Poincaré characteristic (EPC) of a polyconvex subset X of Rd can be evaluated by covering the subset with an auxiliary tessellation, measuring its contribution within each cell of the tessellation and adding all contributions. Two different ways are proposed to define the contribution of a cell to the EPC of X. These contributions turn out to be related by duality formulae. Finally, three applications are given: the measurement of the EPC on adjacent fields, the measurement of the EPC on discretized images and the detection of defects in atomic structures
Local contributions to the Euler-Poincare characteristic of a set
International audienceThe Euler–Poincaré characteristic (EPC) of a polyconvex subset X of Rd can be evaluated by covering the subset with an auxiliary tessellation, measuring its contribution within each cell of the tessellation and adding all contributions. Two different ways are proposed to define the contribution of a cell to the EPC of X. These contributions turn out to be related by duality formulae. Finally, three applications are given: the measurement of the EPC on adjacent fields, the measurement of the EPC on discretized images and the detection of defects in atomic structures