Time course and Ca(2+) dependence of sensitivity modulation in cyclic GMP-gated currents of intact cone photoreceptors.

We determined the Ca(2+) dependence and time course of the modulation of ligand sensitivity in cGMP-gated currents of intact cone photoreceptors. In electro-permeabilized single cones isolated from striped bass, we measured outer segment current amplitude as a function of cGMP or 8Br-cGMP concentrations in the presence of various Ca(2+) levels. The dependence of current amplitude on nucleotide concentration is well described by the Hill function with values of K(1/2), the ligand concentration that half-saturates current, that, in turn, depend on Ca(2+). K(1/2) increases as Ca(2+) rises, and this dependence is well described by a modified Michaelis-Menten function, indicating that modulation arises from the interaction of Ca(2+) with a single site without apparent cooperativity. (Ca)K(m), the Michaelis-Menten constant for Ca(2+) concentration is 857 +/- 68 nM for cGMP and 863 +/- 51 for 8Br-cGMP. In single cones under whole-cell voltage clamp, we simultaneously measured changes in membrane current and outer segment free Ca(2+) caused by sudden Ca(2+) sequestration attained by uncaging diazo-2. In the presence of constant 8Br-cGMP, 15 micro, Ca(2+) concentration decrease was complete within 50 ms and membrane conductance was enhanced 2.33 +/- 0.95-fold with a mean time to peak of 1.25 +/- 0.23 s. We developed a model that assumes channel modulation is a pseudo-first-order process kinetically limited by free Ca(2+). Based on the experimentally measured changes in Ca(2+) concentration, model simulations match experimental data well by assigning the pseudo-first-order time constant a mean value of 0.40 +/- 0.14 s. Thus, Ca(2+)-dependent ligand modulation occurs over the concentration range of the normal, dark-adapted cone. Its time course suggests that its functional effects are important in the recovery of the cone photoresponse to a flash of light and during the response to steps of light, when cones adapt.

In intact cone photoreceptors, a Ca2+-dependent, diffusible factor modulates the cGMP-gated ion channels differently than in rods.

We investigated the modulation of cGMP-gated ion channels in single cone photoreceptors isolated from a fish retina. A new method allowed us to record currents from an intact outer segment while controlling its cytoplasmic composition by superfusion of the electropermeabilized inner segment. The sensitivity of the channels to agonists in the intact outer segment differs from that measured in membrane patches detached from the same cell. This sensitivity, measured as the ligand concentration necessary to activate half-maximal currents, K1/2, also increases as Ca2+ concentration decreases. In electropermeabilized cones, K1/2 for cGMP is 335.5 +/- 64.4 microM in the presence of 20 microM Ca2+, and 84.3 +/- 12.6 microM in its absence. For 8Br-cGMP, K1/2 is 72.7 +/- 11.3 microM in the presence of 20 microM Ca2+ and 15.3 +/- 4.5 microM in its absence. The Ca2+-dependent change in agonist sensitivity is larger in extent than that measured in rods. In electropermeabilized tiger salamander rods, K1/2 for 8Br-cGMP is 17.9 +/- 3.8 microM in the presence of 20 microM Ca2+ and 7.2 +/- 1.2 microM in its absence. The Ca2+-dependent modulation is reversible in intact cone outer segments, but is progressively lost in the absence of divalent cations, suggesting that it is mediated by a diffusible factor. Comparison of data in intact cells and detached membrane fragments from cones indicates that this factor is not calmodulin. At 40 microM 8Br-cGMP, the Ca2+-dependent change in sensitivity in cones is half-maximal at KCa = 286 +/- 66 nM Ca2+. In rods, by contrast, KCa is approximately 50 nM Ca2+. The difference in magnitude and Ca2+ dependence of channel modulation between photoreceptor types suggests that this modulation may play a more significant role in the regulation of photocurrent gain in cones than in rods.

A cyclic-AMP-gated conductance in cochlear hair cells.

The patch clamp technique was used to record cAMP-dependent currents of the guinea pig cochlear hair cell plasma membrane. Data obtained indicate that the channels passing this current are moderately selective for monovalent cations and are effectively blocked by L-cis-diltiazem and reversibly blocked by 1 mM Mg2+ or Ca2+. The single-channel unit conductance estimated in the absence of divalent cations is about 16 pS. The results demonstrate that cyclic nucleotide-dependent channels of cochlear hair cells are virtually identical to the photoreceptor and olfactory ones.

[A new enriched balanced irrigation solution for intraocular surgery (experimental study)]. / Novyi obogashchennyi sbalansirovannyi irrigatsionnyi rastvor dlia vnutriglaznoi khirurgii (éksperimental'noe issledovanie).

Experimental trials of a new intraocular irrigation solution, one of the enriched balanced salines, were carried out. A comprehensive morphologic study has revealed the protector characteristics of lactosol plus (lactosol + taurin) used in the management of irrigation injury to the posterior epithelium of the cornea in the course of experimental intrachamber perfusion. The protective properties of lactosol + taurin, a new enriched balanced saline, are explained by the similarity of its chemical composition to that of the intraocular fluid and by the presence of a stable polyfunctional bioprotector, taurin amino acid.

[The absence of channel structures in the photoreceptor membrane of the rod disc]. / Ob otsutstvii kanal'nykh struktur v fotoretseptornoi membrane diska palochki.

Comparison of electric characteristics of photoreceptor disc and plasma membranes of photoreceptor was made by means of photopotential registration from the adhized to the impregnated by lipids filters photoreceptor discs or isolated rod outer segments. The resistance of the photoreceptor disc membrane is shown to be by three orders of magnitude higher than the resistance of photoreceptor plasma membrane; namely 1-2 MOhm X cm2 versus 1-2 KOhm X cm2. This is the evidence for the absence of channel structures in the disc membrane.