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P Bois

Publications and source records attributed to P Bois.

At least 19 recordsLinked to original sources
PubMed1994

Variable germline and embryonic instability of the human minisatellite MS32 (D1S8) in transgenic mice.

Tandem repeat loci such as minisatellites and trinucleotide repeats frequently show instability. We have investigated mutation at human minisatellite MS32 (locus D1S8) transferred to transgenic mice. Three lines of hemizygous transgenic mice were studied. A single-copy line (110D) was seen to be relatively stable, whilst two multicopy lines showed structural instability of the transgene in pedigrees (lines 109 and 110A). For both these lines, mutant structures were detected as a result of mutation events having occurred in the germline or early embryo. Structural changes seen included gain or loss of minisatellite repeat units (110A and 109), alteration of DNA flanking the minisatellite repeat array (109 only) or deletion of the entire transgene (109 only). This work demonstrates that tandem repeat transgenes can show instability and thus provide additional systems for the analysis of repetitive DNA structural change in mice.

Animals↗
PubMed1994

Effects of two chemically related new Ca2+ channel antagonists, SR33557 (fantofarone) and SR33805, on the L-type cardiac channel.

Fantofarone (SR33557) is a substituted indolizine and SR33805 is a substituted indole. These drugs have been shown to specifically bind to the alpha 1 subunit of the L-type Ca2+ channel at the same site, distinct from those of the classical 1,4-dihydropyridine, phenylalkylamine or benzothiazepine Ca2+ antagonists, but in negative allosteric interaction with them. The present work shows that fantofarone and SR33805 block L-type but not T-type Ca2+ channels in mouse cardiac cells in primary culture. This block is voltage-dependent. Fantofarone and SR33805 are potent Ca2+ channel blockers in depolarized conditions (i.e. at a holding potential of -40 mV) with an EC50 = 1.4 and 4.1 nM, respectively. In polarized conditions (i.e. at a holding potential of -80 mV), SR33805 is a better Ca2+ channel blocker (EC50 = 33 nM) than fantofarone (EC50 = 0.15 microM). Therefore differences in their chemical structures make the blocking action of fantofarone more sensitive to voltage than that of SR33805.

Animals↗
PubMed1994

Characterization of an angiotensin-II-activated chloride current in rabbit sino-atrial cells.

Single sino-atrial cells from rabbit heart were voltage-clamped using the whole-cell configuration of the patch clamp technique under conditions in which most of the ionic and exchange currents known in pacemaker cardiac cells were minimized. Extracellular angiotensin II (AII) activated a time-independent background current. The current-voltage relation of this current showed an outward rectification. The reversal potential was -20 mV with 156 mM Cl- external solution and 54 mM Cl- internal solution. This reversal potential shifted with changes in the transmembrane Cl- gradient in the fashion expected for a chloride current. Anthracene-9-carboxylic acid and diphenylamine-2-carboxylic acid (chloride channels blockers) were found to be effective in blocking the AII-sensitive current. The linear segment of the current-voltage relation can be totally inhibited by the competitive AII-receptor (AT1) antagonist losartan and by the presence of intracellular protein kinase C inhibitor, whereas the outward rectification is only slightly changed. It is concluded that sino-atrial cells should contain protein-kinase-C-sensitive chloride channels which may be activated by angiotensin II via the stimulation of the AT1 receptors.

Angiotensin II↗
PubMed1994

Calcium and melatonin production in dissociated trout pineal photoreceptor cells in culture.

Trout pineal cells maintained in primary culture produce melatonin in high amounts during night time and low amounts during daytime. The dark-induced increase in melatonin production was enhanced, in a dose-dependent manner, by elevating extracellular calcium concentration. Low external calcium concentration reduced nocturnal and diurnal melatonin production. Bay K 8644 increased, in a dose-dependent manner, the dark-induced rise in melatonin output, and this effect was antagonized by nifedipine and verapamil. This suggests a role for the dihydropyridine calcium channels in the regulation of the melatonin output. To confirm this, patch-clamp recordings (whole-cell perforated) were run in a 20 mmol/l barium medium at different holding potentials from -80 mV. A voltage-dependent inward current was activated from -30 mV to +40 mV with a maximal amplitude being observed at 0 mV. This current was drastically increased in the presence of Bay K 8644. Nifedipine inhibited the current both in the absence or in the presence of Bay K 8644. Our results are consistent with the idea that extracellular calcium participates in the control of melatonin secretion by photoreceptor cells. It is suggested that activation of the voltage-dependent L-type channel may modulate this secretion.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1991

Indolizinsulphones. A class of blockers with dual but discriminative effects on L-type Ca2+ channel activity and excitation-contraction coupling in skeletal muscle.

The alpha 1 subunit of the L-type Ca2+ channel plays a dual role in skeletal muscle. It is essential both for L-type Ca2+ channel activity and for the functioning of the voltage-sensor structure that is situated in the triads as a key element for excitation-contraction coupling. This paper shows, with mouse muscle cells in primary culture, that indolizinsulphone SR33557 which has its binding site on the alpha 1 subunit blocks both L-type Ca2+ channel activity and contraction as the more classical 1,4-dihydropyridine blockers. However, unlike other Ca2+ channel blockers, it can pharmacologically discriminate between the two different roles of the alpha 1 subunit. SR33557 inhibition of both contractile and L-type Ca2+ channel activities is very voltage dependent and increases at depolarized potentials. Complete blockade of contraction was observed at low SR33557 concentrations (K0.5 = 20 nM) and was associated with only minor L-type Ca2+ channel blockade (30%). The remaining and major part of the L-type Ca2+ channel activity (70%) was blocked at much higher SR33557 concentrations (K0.5 = 0.6 microM). The results indicate that SR33557 has a much higher affinity for the alpha 1 subunit inserted into the voltage-sensor structure. They also suggest that the voltage-sensor structure, which probably includes most of the total T-tubule alpha 1 subunit, has intrinsic (but relatively small) Ca2+ channel activity.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1991

Evidence for two types of calcium currents in frog cardiac sinus venosus cells.

Two types of calcium currents were recorded in single sinus venosus cells of the frog heart, using the whole-cell patch-clamp technique. The threshold potentials were approximately -65 mV for T-type current and -40 mV for L-type current. The amplitude and time course of T-type current were unaffected by exchanging calcium for barium, while the amplitude of L-type current was increased and its decay slowed. T-type current was neither modified by 10(-6) M nifedipine nor by 10(-7) M isoprenaline in contrast with the effects of these agents on L-type current. T-type current began to inactivate at -90 mV and was fully inactivated at -45 mV. Its steady-state inactivation curve was approximately 35 mV negative to that of L-type current. Overlap of activation and inactivation relationships was present for both T- and L-type currents and was maximal at -57 and -30 mV, respectively. It was concluded that T- and L-type currents can easily be separated by their voltage, kinetic and pharmacological differences. The presence of a high density of T-type current may be correlated to its contribution to the pacemaking function of the sinus venosus cells.

Animals↗
PubMed1991

Conditional immortalization of normal and dysgenic mouse muscle cells by the SV40 large T antigen under the vimentin promoter control.

We have created new mouse muscle cell lines of an immortalized type, expressing normal differentiation at the myotube stage: sarcomeric organization, functional excitation-contraction coupling, and triadic differentiation. The DNA immortalizing recombinant utilizes a deletion mutant of the regulatory region of the human vimentin promoter controlling the expression of a SV40 thermosensitive large T antigen, in which the small t sequence has been deleted. Skeletal mouse replicative myoblasts synthesized predominantly vimentin. After myoblast fusion the vimentin gene is strongly repressed in multinucleated syncytia. Furthermore, the normal activity of the vimentin promoter in myoblasts is increased in the large T antigen-expressing cells. We observed that continuous and rapid division of myoblasts occurs at permissive temperature, suggesting that immortalization is achieved even though the small t antigen is absent. When fusion is induced by changing media conditions, large T antigen expression is totally repressed by the vimentin promoter. When the temperature is elevated to 39 degrees C, the preexisting large T antigen is inactivated. The resulting myotubes from normal mouse differentiate totally normally as indicated by their morphology, ultrastructure, and electrophysiological properties. Mutant (muscular dysgenesis) immortalized cells express the same properties as mutant primary counterparts with no contraction, no slow Ca2+ current, and no triadic differentiation. These immortalized cell lines are potentially very useful for further pharmacology, transplantation, and cell biology studies. The vimentin promoter control of immortalizing recombinant DNA can be used for any mammalian normal and mutant muscle cell lines.

Animals↗
PubMed1990

Isolated cells of the frog sinus venosus: properties of the inward current activated during hyperpolarization.

Single sinus venosus cells from frog, Rana esculenta, were isolated using an enzymic dispersion procedure, involving applications of collagenase and protease. About 40%-60% of the cells showed spontaneous contractions. Isolated cells were studied in the whole-cell configuration. Regenerative action potentials were tetrodotoxin-insensitive and similar to those recorded in multicellular preparations. Hyperpolarizing pulses in the voltage range negative to -50 mV induced the activation of a time-dependent inward current, which was blocked by 4 mM caesium but less affected by barium ions. A lower concentration of caesium (1 mM) exerted a voltage-dependent reduction of the current and decreased the spontaneous pacing rate. The activation range of the hyperpolarization-activated current approximately extended from -50 mV to -110 mV, but varied from cell to cell. A high variability was observed in the behaviour of the activation kinetics. The current had a reversal potential near -20 mV that was shifted positively by increasing the external potassium concentration (from 3 mM to 30 mM) and negatively by reducing the external sodium concentration (from 115 mM to 30 mM). The hyperpolarization-activated inward current of the frog sinus venosus cell appears to be carried by both sodium and potassium ions. It shows electrophysiological properties similar to those of the If current of the mammalian heart. The role of the current in the spontaneous activity is discussed.

Animals↗
PubMed1990

Phenytoin preferentially inhibits L-type calcium currents in whole-cell patch-clamped cardiac and skeletal muscle cells.

The effect of the anticonvulsant diphenylhydantoin (phenytoin) was tested on the inward calcium currents of whole-cell patch-clamped cells from rat and human muscles and from frog atrium. A concentration of 10 microM phenytoin was required to obtain a threshold inhibitory effect and, even with high concentrations (100 microM), the inhibition was not complete. In skeletal muscle (rat and human cells in culture), phenytoin (30 microM) exerted a more potent effect on the high-threshold calcium current (ICa,L inhibition: 53 +/- 6% mean +/- SDn-1) rather than on the low-threshold one (ICa,T inhibition: 16 +/- 10%). Similar results were obtained on dissociated frog atrial cells. These data are to be contrasted with those previously reported on neuronal cells, where specific inhibition of ICa,T was reported. Thus, the action of phenytoin appears to be different in muscle and nerve so that phenytoin does not appear to be a specific inhibitor of ICa,T.

Animals↗
PubMed1987

Characterization of the ionic mechanism responsible for the hyperpolarization-activated current in frog sinus venosus.

Voltage clamp experiments were carried out on the sinus venosus of the frog by means of the double mannitol gap technique. The ionic mechanism underlying the slowly hyperpolarization-activated inward current was investigated by changing the concentration and species of alkali cations and divalent cations in the bathing solution. Adding Rb or Cs in concentration of 10-20 mM to the control solution led to a dose-dependent increase in the inward current, as does elevating the external concentration of K from 2.5 to 25 mM. After the inward current had been nearly suppressed by completely substituting Tris for Na in the external medium, it was partially restored after a subsequent addition of K, Rb or Cs to the Na-free medium. Various alkaline earths or transition metals added to the bathing solution markedly depressed the magnitude of the inward current. This inhibitory effect varied with concentration and nature of divalent cations added. It also depended on the concentration and species of alkali cations present in the external solution. From these observations it was proposed that the conductance responsible for the inward rectification in frog sinus venosus does not discriminate among monovalent cations. The results support the existence of a weak-field-strength site located in the permeation pathway. Divalent cation may exert their inhibitory effect by competing with permeant ions for this site.

Animals↗