Biomedical subjects

Igor Poberaj

Publications and source records attributed to Igor Poberaj.

4 recordsLinked to original sources

PubMed2006

Interparticle potential and drag coefficient in nematic colloids.

Magneto-optic tweezers were used for measurements of liquid-crystal-mediated forces between spherical beads with tangential anchoring in thin nematic samples. Repulsive force, which results from the quadrupolar symmetry of defects around the immersed beads, decreases proportionally to 1/x6, with x being the bead separation. The velocity with which the particles are pushed apart also follows the same separation dependence. We thus find the effective drag coefficient gamma(eff) independent of x for surface-to-surface distances as small as 10% of the bead diameter.

Journal Article↗

PubMed2005

Concentration-dependent staining of lactotroph vesicles by FM 4-64.

Hormones are released from neuroendocrine cells by passing through an exocytotic pore that forms after vesicle and plasma membrane fusion. An elegant way to study this process at the single-vesicle level is to use styryl dyes, which stain not only the membrane, but also the matrix of individual vesicles in some neuroendocrine cells. However, the mechanism by which the vesicle matrix is stained is not completely clear. One possibility is that molecules of the styryl dye in the bath solution dissolve first in the plasma membrane and are then transported into the vesicle by lateral diffusion in the plane of the membrane, and finally the vesicle matrix is stained from the vesicle membrane. On the other hand, these molecules may enter the vesicle lumen and reach the vesicle matrix by permeation through an open aqueous fusion pore. To address these questions, we exposed pituitary lactotrophs to different concentrations of FM 4-64 to monitor the fluorescence increase of single vesicles by confocal microscopy after the stimulation of cells by high K(+). The results show that the membrane and the vesicle matrix exhibit different concentration-dependent properties: the plasma membrane staining by FM 4-64 has a higher affinity in comparison to the vesicle matrix. Moreover, the kinetics of vesicle loading by FM 4-64 exhibited a concentration-dependent process, which indicates that FM 4-64 molecules stain the vesicle matrix by aqueous permeation through an open fusion pore.

Animals↗

PubMed2004

Slow spontaneous secretion from single large dense-core vesicles monitored in neuroendocrine cells.

Hormones are released from cells by passing through an exocytotic pore that forms after vesicle and plasma membrane fusion. In stimulated exocytosis vesicle content is discharged swiftly. Although rapid vesicle discharge has also been proposed to mediate basal secretion, this has not been studied directly. We investigated basal hormone release by preloading fluorescent peptides into single vesicles. The hormone discharge, monitored with confocal microscopy, was compared with the simultaneous loading of vesicle by FM styryl dye. In stimulated vesicles FM 4-64 (4 microM), loading and hormone discharge occurs within seconds. In contrast, in approximately 50% of spontaneously releasing vesicles, the vesicle content discharge and the FM 4-64 loading were slow (approximately 3 min). These results show that in peptide secreting neuroendocrine cells the elementary vesicle content discharge differs in basal and in stimulated exocytosis. It is proposed that the view dating back for some decades, which is that, at rest, the vesicle discharge of hormones and neurotransmitters is similar to that occurring after stimulation, needs to be extended. In addition to the classical paradigm that secretory capacity of a cell is determined by controlling the probability of occurrence of elementary exocytotic events, one will have to consider activity modulation of elementary exocytotic events as well.

Animals↗

PubMed2002

Modeling excess retrieval in rat melanotroph membrane capacitance records.

We have used the patch-clamp technique to monitor changes in membrane capacitance (C(m)) elicited by fast and spatially homogeneous rises in cytosolic calcium concentration ([Ca(2+)](i)) using flash photolysis of NP-EGTA. Average peak [Ca(2+)](i) amplitudes of 20-25 microM triggered three different types of responses in C(m): (i) In 42% of cells, a rise in [Ca(2+)](i) activated a monotonic increase in C(m) followed by a slow decline to resting values; (ii) In 30% of cells, the rise in C(m) was clearly characterized by two dynamic components, consisting of a rapid and a slow exo-endocytosis cycle; (iii) In 28% of cells, after the initial rapid rise in C(m), endocytosis exhibited excess retrieval that was characterized by a decline in C(m) below resting C(m). The aim of this work is to develop a unified mathematical model with a minimum number of parameters that would describe all the observed types of responses. Three models were considered: Model A, a model with a single component of exo-endocytosis cycle; model B, a model consisting of a sum of two independent dynamic components; and model C, a model in which, in addition to the two dynamic components as in model B, excess retrieval due to a lipid flow through the reversal closing of the fusion pore during the rapid component of exo-endocytosis cycle was considered. The results show that the latter model describes all the types of responses in C(m) recorded in rat melanotrophs. The association of excess retrieval exclusively with the rapid, but not the slow, exocytosis indicates that some fusing vesicles mediate a lipidic flux during the reversal closing of the fusion pore, whereas those entering the slow phase of exocytosis may fuse with the plasma membrane completely and are retrieved by other endocytic machinery, independent of the lipid flow that might have occurred as the fusion pore opened permanently.

Animals↗