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L Lagnado

Publications and source records attributed to L Lagnado.

26 records · Page 2Linked to original sources
PubMed1992

Calcium homeostasis in the outer segments of retinal rods from the tiger salamander.

1. The processes regulating intracellular calcium in the outer segments of salamander rods have been investigated. The main preparation used was the isolated rod loaded with the Ca(2+)-sensitive photoprotein aequorin, from which outer segment membrane current and free [Ca2+]i could be recorded simultaneously. Two other preparations were also used: outer segment membrane current was recorded from intact, isolated rods using a suction pipette, and from detached outer segments using a whole-cell pipette. 2. Measurements of free intracellular [Ca2+] in Ringer solution were obtained from two aequorin-loaded rods. Mean [Ca2+]i in darkness was 0.41 microM, and after a bright flash [Ca2+]i fell to below detectable levels ( < 0.3 microM). No release of intracellular Ca2+ by a bright flash of light could be detected ( < 0.2 microM). 3. Application of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) caused an increase in the size of the light-sensitive current and a rise in [Ca2+]i, but application of IBMX either when the light-sensitive channels had been closed by a bright light or in the absence of external Ca2+ caused no detectable rise in [Ca2+]i. It is concluded that IBMX increases [Ca2+]i by opening light-sensitive channels, and does not release Ca2+ from stores within the outer segment. 4. Removal of external Na+ caused a rise in [Ca2+]i to around 2 microM and completely suppressed the light-sensitive current. 5. The Na(+)-Ca2+, K+ exchange current in aequorin-loaded rods was activated in first-order manner by internal free calcium, with a mean Michaelis constant, KCa, of 1.6 microM. 6. The KCa of the Na(+)-Ca2+, K+ exchange was increased by elevating internal [Na+]. 7. The Michaelis relation between [Ca2+]i and the activity of the Na(+)-Ca2+, K+ exchange was used to calculate the change in [Ca2+]i occurring during the response to a bright light. In aequorin-loaded rods in Ringer solution the mean change in free [Ca2+]i after a bright flash was 0.34 microM. In these rods 10% of the dark current was carried by Ca2+. 8. Most of the calcium entering the outer segment was taken up rapidly and reversibly by buffer systems. The time constant of equilibration between free and rapidly bound Ca2+ was less than 20 ms. No slow component of calcium uptake was detected. 9. Two components of calcium buffering could be distinguished in the outer segments of aequorin-loaded rods.(ABSTRACT TRUNCATED AT 400 WORDS)

1-Methyl-3-isobutylxanthine↗
PubMed1991

Net charge transport during sodium-dependent calcium extrusion in isolated salamander rod outer segments.

The light-sensitive current and the current associated with the extrusion of internal Ca2+ in exchange for external Na+ have been recorded from detached rod outer segments from the salamander retina by the use of the whole-cell voltage clamp technique. No significant current-carrying mechanisms are present in the outer segment membrane apart from the light-sensitive conductance and the Na:Ca,K exchange, and exchange currents can therefore be recorded directly without the use of subtraction procedures or pharmacological blockers. The charge moved by the exchange was studied by loading outer segments with a known amount of calcium and then recording the exchange current on return to a Na(+)-containing solution. Calcium is not sequestered to any significant extent in a slowly exchanging internal store, as the charge recovered is unaffected if admission of the Na(+)-containing solution is delayed for 40 s. The number of charges flowing into the cell in exchange for each Ca2+ ion extruded was found not to deviate significantly from one over a wide range of ionic conditions and membrane potentials. These results show that the stoichiometry of the exchange is fixed over a wide range of conditions, and that the size of the inward exchange current is therefore directly proportional to the rate of Ca2+ efflux through the carrier.

1-Methyl-3-isobutylxanthine↗
PubMed1990

The effects of quinidine on sodium-dependent calcium efflux in isolated rod photoreceptors of the salamander retina.

The effect of quinidine on the membrane current generated by the Na:Ca, K exchange has been investigated in the outer segment of isolated rod photoreceptors from the retina of the larval tiger salamander. The inward exchange current associated with the efflux of Ca2+ was selectively recorded by introducing a Ca2+ load through the light-sensitive channels, and then shutting these channels with a bright light. Quinidine (20-1000 microM) reduced the magnitude of the exchange current and slowed its decay during the removal of a Ca2+ load. Quinidine did not alter the form of the relation between the exchange current and the total concentration of exchangeable calcium remaining within the outer segment. [Ca]T, showing that it does not change the affinity of the exchange mechanism for internal Ca2+. The relation between exchange current inhibition and the quinidine concentration could be described by a simple Michaelis relation with a Ki of 287 microM and a maximum inhibition of 50%. The incomplete block of the Na:Ca, K exchange current by quinidine shows that it does not act by simple competition with external Na+, and suggests that the inhibition of the exchange by quinidine may be non-specific.

1-Methyl-3-isobutylxanthine↗
PubMed1989

Extrusion of calcium from rod outer segments is driven by both sodium and potassium gradients.

Calcium is transported across the surface membrane of both nerve and muscle by a Na+-dependent mechanism, usually termed the Na:Ca exchange. It is well established from experiments on rod outer segments that one net positive charge enters the cell for every Ca2+ ion extruded by the exchange, which is generally interpreted to imply an exchange stoichiometry of 3 Na+:1 Ca2+. We have measured the currents associated with the operation of the exchange in both forward and reversed modes in isolated rod outer segments and we find that the reversed mode, in which Ca2+ enters the cell in exchange for Na+, depends strongly on the presence of external K+. The ability of changes in external K+ concentration ([K+]o) to perturb the equilibrium level of [Ca2+]i indicates that K+ is co-transported with calcium. From an examination of the relative changes of [Ca2+]o, [Na+]o, [K+]o and membrane potential required to maintain the exchange at equilibrium, we conclude that the exchange stoichiometry is 4 Na+:1 Ca2+, 1 K+ and we propose that the exchange should be renamed the Na:Ca, K exchange. Harnessing the outward K+ gradient should allow the exchange to maintain a Ca2+ efflux down to levels of internal [Ca2+] that are considerably lower than would be possible with a 3 Na+:1 Ca2+ exchange.

Algorithms↗
PubMed1988

Ion transport by the Na-Ca exchange in isolated rod outer segments.

The inward membrane current generated by the coupled exchange of external sodium for internal calcium has been investigated in isolated rod outer segments. The exchange rate is sensitive to voltage, with a reduction by a factor of e occurring for a 70-mV depolarization in normal Ringer's solution. The voltage sensitivity is not a constant property of the exchange, as it is reduced by an increase in external Na+ or by the removal of external Ca2+, Mg2+, or K+. Changes in membrane potential do not appear to affect the affinity of the exchange mechanism for internal Ca2+, but hyperpolarization increases the affinity for external Na+. When the external Na+ concentration is raised sufficiently to saturate the exchange mechanism, the voltage sensitivity is no longer apparent. We propose that the voltage dependence of the exchange is due to the external Na+-binding site being sensitive to membrane potential, perhaps because it is located within the membrane electric field.

Animals↗
PubMedDate not supplied

Visual processing: the devil is in the details.

Ganglion cells convey information from the retina back to the brain. Recent experiments have examined how ganglion cell receptive fields are assembled from many incoming signals.

Animals↗