87 research outputs found
Inactivation of single Ca2+ channels in rat sensory neurons by extracellular Ca2+
No full text1. Single Ca2+ channels conducting 20 mM Ba2+ from adult rat dorsal root ganglion cells were characterized using the two-electrode patch-clamp technique configuration. 2. Channels demonstrating specific characteristics of conductance, voltage dependence and dihydropyridine sensitivity were classified as high-threshold or L-type Ca2+ channels. 3. Mean single-channel current in 20 mM Ba2+ did not show inactivation, but inactivation occurred when using Ca2+ as a permeating ion. 4. Stimulus protocols were delivered alternately in the cell-attached and whole-cell electrode, while recording single-channel activity and total Ca2+ current simultaneously. 5. A mean single-channel Ba2+ current from a stimulated patch did not show inactivation. However, stimulation of a physiological whole-cell Ca2+ current induced a marked inactivation of mean single-channel Ba2+ current. 6. Complete Ca2+ current block by the addition of 200 microM Cd2+ in the external solution removed single-channel inactivation in patches stimulated through a whole-cell electrode
Characterization of Ca2+ transients induced by intracellular photorelease of InsP3 in mouse ovarian oocytes.
No full textCa2+ transients (measured with Fluo-3) were induced in single mouse ovarian oocytes by photolytic liberation of InsP3. The time course of cytosolic Ca2+ changes induced in this way is composed of distinct phases: upstroke, fast decline, slow declining plateau and fast decline to rest level. All the phases reflect mainly intracellular redistributions of the ion and not influx, since they are not strongly dependent on external Ca2+ or on changes in transmembrane potential. Often sustained Ca2+ oscillations followed the first InsP3-induced Ca2+ transient. These persisted for several minutes in the absence of external Ca2+. The initial rate of Ca2+ rise and the delay between the InsP3 stimulus and Ca2+ upstroke are correlated with the amount of liberated InsP3. A second InsP3 stimulation, applied during the plateau, causes only small Ca2+ elevations, lacking the upstroke phase. A second, full sized, transient could be elicited only after a complete return to the basal level. Vanadate, applied intracellularly, appeared to inhibit the re-uptake phase into the stores, stabilizing the plateau level. The present observations suggest that in mouse oocytes the InsP3-sensitive stores provide only a small and graded Ca2+ release which may then act as a trigger for a more substantial Ca(2+)-induced Ca2+ release (CICR) process
Neurofilament-negative neurones exhibit HVA Ca2+ currents with faster inactivation kinetics
No full textThe whole cell configuration of the patch clamp technique was used to study the biophysical and pharmacological properties of voltage activated Ca2+ channel currents on hippocampal neurones cultured in the presence or absence of foetal calf serum. In the presence of serum cells were intensively immunostained with neurofilament (NF) antibodies. In the absence of serum, cells were non-immunoreactive; they were identified as neurones by their ability to generate action potentials. NF negative cells still expressed both high (HVA) and low (LVA) voltage activated Ca2+ channels. However, in comparison to NF positive neurones, HVA Ca2+ currents present in NF negative neurones had a faster inactivation kinetics
“Superstition ain’t the way”. The optimism of the conspiracy theorist
No full textThis article addresses the topic of conspiracy theories from a rhetorical and philosophical perspective through a recent case study: the Netflix docuseries The Ancient Apocalypse by Graham Hancock1. In this TV series, the host – the author – hypothesizes a conspiracy by academic archaeology, which does not accept the idea of the existence of a highly technically developed ancient civilization that disappeared before the last ice age. The reason for this resistance on the part of archaeologists would be their desire to maintain a position of power and prestige that they would have to give up if they accepted a paradigm shift. We will use Hancock’s speeches as a textual corpus to be analyzed from a rhetorical point of view, and we will follow the philosophical hypothesis that conspiracy theories should be understood through the phenomenon of superstition (which is different from magic, § 1). We devote a paragraph to each technical proof used to achieve persuasion. First, éthos (§ 2), which focuses on the construction of the speaker as an outsider, credible because he is inconvenient. We then analyze the logos (§ 3), characterized by fallacies, ambivalent logic, and the mythological narrative as historical evidence. Finally (§ 4), we focus on pathos, which exhibits a form of detachment and a disposition to inaction
Neurotrophin-3 promotes the survival of oligodendrocyte precursors in embryonic hippocampal cultures under chemically defined conditions
No full textEmbryonic rat hippocampal cells were cultured in basal medium with or without addition of the neurotrophin NT-3. After culturing in these extreme conditions, the effects of NT-3 on the neuronal and on the glial components were assessed. Neurons survived even in the absence of NT-3 but failed to reach terminal differentiation. On the other hand, NT-3 promoted the survival but not the proliferation and/or the differentiation of oligodendrocytes precursors present in the same culture, an effect that was reversed by the addition of neutralizing antibodies against NT-3. Type I or II astrocytes were not affected by NT-3. These results reinforce the role for NT-3 in oligodendrocyte lineage development and allow to dissect the roles of this neurotrophin in survival and in proliferation/differentiation of oligodendrocytes
Neurofilaments-negative neurons exhibit HVA Ca2+ currents with faster inactivation kinetics
No full textP2y purinoceptors in normal NIH 3T3 and in NIH 3T3 overexpressing c-ras
No full textThe ability of purinergic agonists to induce Ca2+ responses has been tested in two lines of murine fibroblasts: normal NIH 3T3 fibroblasts and NIH 115.14, a clone expressing high levels [1] of the c-ras protooncogene. Both kinds of cells are responsive to ATP in the range 1 microM-1 mM; ADP and ATP gamma S are almost as potent as ATP, while AMP is unable to elicit a response. Ca2+ measurements performed in single cells by image analysis show great variability among cells but in each individual responding cell the Ca2+ rise occurs in an all-or-none fashion. The transient Ca2+ response does not depend on influx from the extracellular medium. Electrophysiological experiments reveal the activation of an outward current (at -50 mV) by ATP, probably due to Ca(2+)-activated K+ channels, confirming the absence of a substantial Ca2+ influx. Finally, stimulation by ATP produces a small but significant increase in the production of inositol phosphates. These results indicate that these cell lines possess purinergic receptors which are not integral membrane channels and which are coupled to InsP3 formation and may be therefore classified as P2Y
Cytosolic calcium responses induced by photolytic release of 1,4,5-inositol trisphosphate in single human fibroblasts.
No full textWe have used the whole cell technique to microinject human fibroblasts with either 1,4,5-inositol trisphosphate (InsP3) or 'caged' InsP3, in order to study the mechanisms of transmembrane signalling related to mitogenic stimulations. Cytosolic Ca2+ elevations in response to 1,4,5 InsP3 diffusing from the patch pipette were difficult to detect, while 1,4,5 InsP3, photoreleased after loading the cell with its inactive precursor, was capable of generating not only a single cytosolic Ca2+ rise but sometimes triggered an oscillatory calcium response, similar to that often observed under mitogenic stimulation. We estimated that less than 100 nM InsP3 was sufficient to generate Ca2+ responses. The Ca2+ rise produced by the photoreleased InsP3 could fully activate the K+ channels present in the plasma membrane of human fibroblasts
[Ca2+]i recordings and the inactivation of the high-voltage activated Ca2+ currents in the adult rat sensory neuron.
No full textFast, single cell, measurement of the average cytosolic [Ca2+]i with the Fura-2 technique suggests that the depolarization induced [Ca2+]i rise is entirely due to entry through the voltage-activated Ca2+ channels. Involvement of a Ca(2+)-induced Ca(2+)-release process is not evident. Under physiological cytosolic buffering the current-induced [Ca2+]i rise persists for seconds and decays exponentially (tau = 7 s). Analysis of the [Ca2+]i changes during two-pulse protocols indicates that the purely voltage-dependent inactivation of the high voltage-activated (HVA) channels, in the range -80/+70 mV, is a slow process (0.2-1 s) which removes at most 40% of the current. On the contrary, Ca(2+)-dependent inactivation acts in a fast way and it is therefore responsible for the fast inactivating phase of the current; this phase disappears under sustained [Ca2+]i loads, and reappears when redistribution of free Ca2+ takes place. A suitable correction may be devised to compensate for the Ca(2+)-dependent inactivation
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