PubMed Health⌕ Search

Biomedical subjects

A Picones

Publications and source records attributed to A Picones.

12 recordsLinked to original sources
PubMed2001

Unitary conductance variation in Kir2.1 and in cardiac inward rectifier potassium channels.

Kir2.1 (IRK1) is the complementary DNA for a component of a cardiac inwardly rectifying potassium channel. When Kir2.1 is expressed in Xenopus oocytes or human embryonic kidney (HEK) cells (150 mM external KCl), the unitary conductances form a broad distribution, ranging from 2 to 33 pS. Channels with a similarly broad distribution of unitary conductance amplitudes are also observed in recordings from adult mouse cardiac myocytes under similar experimental conditions. In all three cell types channels with conductances smaller, and occasionally larger, than the ~30 pS ones are found in the same patches as the ~30 pS openings, or in patches by themselves. The unitary conductances in patches with a single active channel are stable for the durations of the recordings. Channels of all amplitudes share several biophysical characteristics, including inward rectification, voltage sensitivity of open probability, sensitivity of open probability to external divalent cations, shape of the open channel i-V relation, and Cs(+) block. The only biophysical difference found between large and small conductance channels is that the rate constant for Cs(+) block is reduced for the small-amplitude channels. The unblocking rate constant is similar for channels of different unitary conductances. Apparently there is significant channel-to-channel variation at a site in the outer pore or in the selectivity filter, leading to variability in the rate at which K(+) or Cs(+) enters the channel.

Animals↗
PubMed2000

Developmental switch in excitability, Ca(2+) and K(+) currents of retinal ganglion cells and their dendritic structure.

In the retina of teleost fish, continuous neuronal development occurs at the margin, in the peripheral growth zone (PGZ). We prepared tissue slices from the retina of rainbow trout that include the PGZ and that comprise a time line of retinal development, in which cells at progressive stages of differentiation are present side by side. We studied the changes in dendritic structure and voltage-dependent Ca(2+), Na(+), and K(+) currents that occur as ganglion cells mature. The youngest ganglion cells form a distinct bulge. Cells in the bulge have spare and short dendritic trees. Only half express Ca(2+) currents and then only high-voltage-activated currents with slow inactivation (HVAslow). Bulge cells are rarely electrically excitable. They express a mixture of rapidly inactivating and noninactivating K(+) currents (IKA and IKdr). The ganglion cells next organize into a transition zone, consisting of a layered structure two to three nuclei thick, before forming the single layered structure characteristic of the mature retina. In the transition zone, the dendritic arbor is elaborately branched and extends over multiple laminae in the inner plexiform layer, without apparent stratification. The arbor of the mature cells is stratified, and the span of the dendritic arbor is well over five times the cell body's diameter. The electrical properties of cells in the transition and mature zones differ significantly from those in the bulge cells. Correlated with the more elaborate dendritic structures are the expression of both rapidly inactivating HVA (HVAfast) and of low-voltage-activated (LVA) Ca(2+) currents and of a high density of Na(+) currents that renders the cells electrically excitable. The older ganglion cells also express a slowly activating K(+) current (IKsa).

Animals↗
PubMed1999

A developmental time line in a retinal slice from rainbow trout.

The retina in teleost fish continues to grow throughout much of the life of the animal, in part by the continuing differentiation of new tissue at the retinal margin, an area termed the peripheral growth zone (PGZ) (Lyall, Q J Micros Sci, 1957:98:101-110). We have developed a retinal slice preparation--including the PGZ--from juvenile rainbow trout (Onchorynchus mykiss), a species in which retinal growth is rapid and the PGZ is correspondingly pronounced. The PGZ slice preparation contains a time line of retinal development, with cells at different stages of maturation present side by side. We present evidence that the birth sequence of the various retinal cell types in the PGZ recapitulates the sequence during embryonic development. We also report data on the rate of growth of the PGZ in juvenile trout in vivo. Finally, we have used the PGZ slice preparation to make whole-cell voltage clamp recordings from individual retinal GCs at both early and late stages of maturation. We report that the amplitude of delayed rectifier and A-type potassium currents increases during GC maturation.

Animals↗
PubMed1997

Localization and release of 5-hydroxytryptamine in the crayfish eyestalk

The content and regional distribution of 5-hydroxytryptamine (5-HT) in the crayfish eyestalk was determined by high-performance liquid chromatography. Levels of the 5-HT precursors l-tryptophan (L-TRP) and 5-hydroxytryptophan (5-OH-TRP), and of three metabolites, 5-hydroxytryptophol (5-HTPH), N-acetylserotonin (NA-5-HT) and 5-hydroxy-indole-3-acetic acid (5-HIAA), were also determined. The total content of 5-HT in the eyestalk was 95.4+/-49.3 pg mg-1 wet mass (mean +/- s.d., N=55) while the specific content was 9.6+/-4.9 fmol microg-1 protein (mean +/- s.d. N=5). 5-HT was present in all four ganglia of the eyestalk. The highest proportion was found in the medulla terminalis (40.2 %) and the lowest in the retina lamina ganglionaris (9.9 %), which also had the lowest specific content. Conversely, the highest specific content of L-TRP was in the retina lamina ganglionaris. 5-HT biosynthesis and metabolism were explored in isolated eyestalks. The monoamine oxidase blocker pargyline, at concentrations between 0.8 and 10 mmol l-1, elicited a dose-dependent increase in 5-HT content. The biosynthesis of 5-HT in the crayfish eyestalk is suggested by the presence of its immediate precursor (5-OH-TRP) and by the suppression of 5-HT synthesis induced by m-hydroxybenzyl-hydrazine (m-HBH), a blocker of 5-OH-TRP decarboxylase. The presence of immunopositive cell bodies and axons was demonstrated using an anti-5-HT antiserum. 5-HT-like immunopositivity was detected in various regions of the eyestalk. Efferent immunopositive axons were also identified in the optic nerve, and these may have originated in the protocerebral lobe of the supraoesophageal ganglion. The branchings of these axons were profusely distributed in the neuropile of the medulla terminalis. A basal level release of 5-HT was detected in isolated eyestalks. The amount recovered was increased two-to threefold after blocking 5-HT uptake with fluoxetine (1 micromol l-1). Incubation of eyestalks in solutions containing a high K+ concentration (80 mmol l-1) released 5-HT. Electrical stimulation of the optic nerve released 5-HT as a function of the intensity of stimulation. Both the basal and evoked release were suppressed by lowering the Ca2+ concentration in the medium. These observations support a role for 5-HT as a neurotransmitter or neuromodulator in the crayfish eyestalk.

Journal Article↗
PubMed1995

Spontaneous, ligand-independent activity of the cGMP-gated ion channels in cone photoreceptors of fish.

1. We studied the electrical conductance of membrane patches detached from the outer segment of single cone photoreceptors isolated from striped bass retina. 2. Only a single class of ion channels exists in the plasma membrane of the cone outer segments; they are gated by cytoplasmic cGMP and select cations over anions, but distinguish poorly among cations. In the absence of added cGMP and of divalent cations, however, membrane patches detached from the outer segments exhibit a small conductance that ideally selects cations over anions, but distinguishes poorly between Na+ and Li+. 3. The cGMP-independent conductance does not arise from the effect of residual cGMP that may remain associated with the detached membrane, because treatment of the patch with cGMP-specific phosphodiesterase does not affect this conductance. 4. The cGMP-independent conductance is pharmacologically indistinguishable from that activated by cGMP. Ca2+ and L-cis-diltiazem block both conductances at comparable concentrations and with similar quantitative characteristics. 5. We analysed the noise of Ca(2+)- or L-cis-diltiazem-dependent macroscopic currents both in the presence and in the absence of cGMP. In the presence of cGMP, the power density spectrum of the noise is well fitted by the sum of two Lorentzian components. The same function with similar corner frequencies fits the noise of the cGMP-independent currents. However, the total power in the current fluctuations is smaller in the absence of cGMP than in its presence; also, the ratio of the zero frequency asymptotes of the low over the high frequency components, S1(0)/Sh(0), is larger in the absence of cGMP than in its presence.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Permeability and interaction of Ca2+ with cGMP-gated ion channels differ in retinal rod and cone photoreceptors.

We studied the ionic permeability of cGMP-dependent currents in membrane patches detached from the outer segment of retinal cone and rod photoreceptors. Reversal potentials measured in membranes exposed to symmetric Na+ but with varying cytoplasmic Ca2+ concentrations reveal that the permeability ratio, PCa/PNa, is higher in the cGMP-gated channels of cones (7.6 +/- 0.8) than in those of rods (3.1 +/- 1.0). Ca2+ blocks both channels in a voltage-dependent manner. At any Ca2+ concentration, the channel block is maximal near the ionic reversal potential. The maximal block is essentially identical in channels of cones and rods with respect to its extent and voltage and Ca2+ dependence. The Ca2+ block is relieved by voltage, but the features of this relief differ markedly between rods and cones. Whereas the Boltzmann distribution function describes the relief of block by hyperpolarizing voltages, any given voltage is more effective in relieving the Ca2+ block in cones than in rods. Similarly, depolarizing voltages more effectively relieve Ca2+ block in cones than in rods. Our results suggest that channels contain two binding sites for Ca2+, one of which is similar in the two receptor types. The second site either interacts more strongly with Ca2+ than the first one or it is located differently in the membrane, so as to be less sensitive to membrane voltage. The channels in rods and cones differ in the features of this second site. The difference in Ca2+ permeability between the channels is likely to result in light-dependent changes in cytoplasmic Ca2+ concentration that are larger and faster in cones than in rods. The functional differences between channels, therefore, may be critically important in explaining the differences in the phototransduction signal of the two photoreceptor types.

Ambystoma↗
PubMed1994

Differences in transduction between rod and cone photoreceptors: an exploration of the role of calcium homeostasis.

Rod and cone photoreceptors respond to light with distinct sensitivity and kinetics. Recent biochemical and electrophysiological studies demonstrate that the enzymes of the phototransduction cascade are similar, but not identical, in these two photoreceptor types. In contrast, light or voltage stimulation generates changes in the cytoplasmic concentration of Ca2+ in the outer segment that are far larger and faster in cones than in rods. This distinction reflects rod-cone differences in each of the elements that control Ca2+ homeostasis: cell volume, the rate of Ca2+ clearance from the outer segment, the cytoplasmic Ca2+ buffering, and the Ca2+ influx through cGMP-gated ion channels.

Animals↗
PubMed1994

Analysis of fluctuations in the cGMP-dependent currents of cone photoreceptor outer segments.

We measured cGMP-dependent currents, under voltage clamp, in membrane patches detached from the outer segment of single-cone photoreceptors isolated from the retina of striped bass. We analyzed the variance of the current about its mean and the spectral density distribution of the current fluctuations. From the analysis of variance, we determined that the cGMP-gated channels increase their probability of opening with increasing cGMP up to a maximum value of 0.87 +/- 0.03. The dependence on cGMP of the probability of opening is well described by a Hill equation with Km = 60.2 +/- 3.7 microM and n = 2.33 +/- 0.32 at -50 mV. At the same voltage, the spectral density distribution is well fit by the sum of two Lorentzians with corner frequencies at 26 +/- 18 and 318 +/- 58 Hz. The single-channel conductance calculated from the current noise by two different methods suggests that the most frequently occupied conductance state has an amplitude of about 18 pS.

Analysis of Variance↗
PubMed1992

Permeation and interaction of monovalent cations with the cGMP-gated channel of cone photoreceptors.

We measured the ion selectivity of cGMP-dependent currents in detached membrane patches from the outer segment of cone photoreceptors isolated from the retina of striped bass. In inside-out patches excised from either single or twin cones the amplitude of these currents, under symmetric ionic solutions, changed with the concentration of cGMP with a dependence described by a Hill equation with average values, at +80 mV, of Km = 42.6 microM and n = 2.49. In the absence of divalent cations, and under symmetric ionic solutions, the I-V curves of the currents were linear over the range of -80 to +80 mV. The addition of Ca altered the form of the I-V curve to a new function well described by an empirical equation that also describes the I-V curve of the photocurrent measured in intact photoreceptors. The monovalent cation permeability sequence of the cGMP-gated channels in the absence of divalent ions was PK > PNa = PLi = PRb > PCs (1.11 > 1.0 = 0.99 = 0.96 > 0.82). The conductance selectivity sequence at +80 mV was GNa = GK > GRb > GCs > GLi (1.0 = 0.99 > 0.88 > 0.74 > 0.60). The organic cations tetramethylammonium (TMA) and arginine partially blocked the current, but the larger ion, arginine, was permeant, whereas the smaller ion, TMA, was not. The amplitude of the outward current through the channels increased with the concentration of monovalent cations on the cytoplasmic membrane surface, up to a saturating value. The increase was well described by the adsorption isotherm of a single ion binding site within the channel with average binding constants, at +80 mV, of 104 mM for Na and 37.6 mM for Li. By assuming that the ion channel contains a single ion binding site in an energy trough separated from each membrane surface by an energy barrier, and using Eyring rate theory, we simulated I-V curves that fit the experimental data measured under ionic concentration gradients. From this fit we conclude that the binding site interacts with one ion at a time and that the energy barriers are asymmetrically located within the membrane thickness. Comparison of the quantitative features of ion permeation and interaction between the cGMP-gated channels of rod and cone photoreceptors reveals that the ion binding sites are profoundly different in the two types of channels. This molecular difference may be particularly important in explaining the differences in the transduction signal of each receptor type.

Animals↗
PubMed1990

Modulation of crayfish retinal sensitivity by 5-hydroxytryptamine.

The responsiveness of crayfish retinal photoreceptors to light was enhanced by exposure to 5-hydroxytryptamine (5-HT), either following injection into whole animals or following topical application to isolated eyestalks or retinas. The effect was measured as an increment in the amplitude of the receptor potential, and was dose-dependent in the range 10(-6)-10(-3) mol l-1 (injected as a 0.1 microliter dose in intact animals). It was more pronounced at low levels of illumination and was reversibly blocked by methysergide. The enhancement was a consequence of a dual effect: (a) retraction of the proximal pigment granules within the photoreceptors, with a corresponding increase in the light-admittance function of the retina; and (b) a direct effect, facilitating a membrane conductance increase which mediated the generation of the receptor potential. A set of axons in the lamina ganglionaris with a 5-HT-like immunoreactivity was found in the vicinity of the photoreceptor axons. 5-HT antagonists were capable of blocking the physiological retraction of pigment granules in photoreceptors at night, suggesting that 5-HT acts as a modulator during the nocturnal phase of the circadian cycle in the crayfish retina.

Animals↗
PubMed1988

Effect of light deprivation on the ERG responses of taurine-deficient rats.

The decrease of taurine levels in the retina of taurine-depleted rats treated with guanidinoethane sulfonate (GES) reduced the amplitude of the a- and b-waves of the electroretinogram, registered as a function of the log relative intensity stimulus. The effect was observed either in rats exposed to a light-darkness cycle or to continuous darkness; this effect was somewhat more pronounced in retinas of rats exposed to light. Values obtained from the Naka-Rushton equation for the V-log I curve for the a-wave showed that the half-saturating intensity was unchanged but Vmax was decreased by GES treatment. Implicit times for the b-wave delayed whereas those for the a-wave were unaffected by the treatment.

Animals↗