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Biomedical subjects

P B Detwiler

Publications and source records attributed to P B Detwiler.

6 recordsLinked to original sources

The influence of arrestin (48K protein) and rhodopsin kinase on visual transduction.

The shutoff of the phototransduction cascade in retinal rods requires the inactivation of light-activated rhodopsin. The underlying mechanisms were studied in functionally intact detached rod outer segments by testing the effect of either sangivamycin, an inhibitor of rhodopsin kinase, or phytic acid, an inhibitor of 48K protein binding to phosphorylated rhodopsin, on light responses recorded in whole-cell voltage clamp. The results suggest that isomerized rhodopsin is inactivated fully by multiple phosphorylation and that the binding of 48K protein accelerates recovery by quenching partially phosphorylated rhodopsin. Higher concentrations of sangivamycin cause changes in the light response that cannot be explained by selective inhibition of rhodopsin kinase and suggest that other protein kinases are needed for normal rod function.

3',5'-Cyclic-GMP Phosphodiesterases

Some unresolved issues in the physiology and biochemistry of phototransduction.

A number of recent review articles have discussed what is known about the events responsible for generating the electrical light response in vertebrate photoreceptors. The similarity of the material covered and the unanimity of the conclusions drawn have given rise to the popular, but false, impression that visual transduction is understood fully. The purpose of the present review is to dispell this notion by focusing on some of the unresolved issues.

Animals

Electrical coupling between cones in turtle retina.

1. The electrical coupling between cones of known spectral sensitivity in the peripheral part of the turtle's retina was studied by passing current through a micro-electrode inserted into one cone and recording with a second micro-electrode inserted into a neighbouring cone. 2. Spatial sensitivity profiles were determined by recording flash responses to a long narrow strip of light which was moved across the impaled cones in orthogonal directions. These measurements gave both the length constant lambda of electrical spread in the cone network and the separation of the two cones. 3. The cone separation determined from the spatial profiles agreed closely with that measured directly by injecting a fluorescent dye into two cones. 4. The length constant lambda varied from 18 to 39 micron with a mean of 25 micron for red-sensitive cones and 26 micron for green-sensitive cones. 5. The majority of cone pairs studied were electrically coupled provided they had the same spectral sensitivity and were separated by less than 60 micron: thirty-two out of thirty-six red-red pairs, two out of two green-green pairs, none out of eight red-green pairs: no blue cones were observed. 6. The strength of electrical coupling was expressed as a mutual resistance defined as the voltage in one cell divided by the current flowing into the other. Mutual resistances decreased from a maximum value of about 30 M omega at separations close to zero to 0.2 M omega, the lower limit of detectable coupling at separations of about 60 micron. Mutual resistances were always positive and were independent of which cell was directly polarized. The coupling seemed to be ohmic and any rectification or non-linearity probably arose in the cone membranes rather than in the coupling resistances. 7. The results were analysed in terms of the Lamb & Simon (1977) theories of square and hexagonal lattices, which approximate to the continuous sheet model except in the case of the cone to which current is applied. 8. The total membrane resistance of a single cone was estimated as 100--300 M omega and the connecting resistances as 100 M omega for a square array and 170 M omega for a hexagonal array. The input resistance of a cone in the network was 25--50 M omega. Lower values were often obtained but may be due to injury by the micro-electrodes. 9. The time constant of an isolated cone was estimated as about 20 msec and the capacity as about 100 pF. 10. Discrepancies between experimental findings and theoretical predictions of the hexagonal or square array models were tentatively attributed to an overestimate of lambda resulting from light scattering.

Animals

A surprising property of electrical spread in the network of rods in the turtle's retina.

Flashing a localised stimulus onto a turtle's retina produces an intracellular potential wave which spreads through electrical connections from illuminated to unilluminated photoreceptors. The response in unilluminated rods (but not in cones) becomes faster as the distance from the source increases, perhaps because voltage-dependent permeability changes in the rod membrane make the coupled network behave like a high-pass filter.

Animals

Multiple light-evoked conductance changes in the photoreceptors of Hermissenda crassicornis.

1. Light responses were recorded from the photoreceptors of Hermissenda crassicornis. The response to a flash is a complex potential change involving an initial depolarization, a hyperpolarization, and a depolarizing tail. None of the phases of the response are due to synaptic interactions.2. Polarization of the membrane by extrinsic current indicates that three separate conductance changes are associated with the response. The initial depolarization and hyperpolarization are accompanied by conductance increases and the tail with a conductance decrease. The initial depolarization has a positive reversal potential and the hyperpolarizing and tail phase have a reversal voltage more negative than resting potential.3. The different processes that give rise to the conductance changes have similar spectral sensitivities but are affected unequally by light adaptation. Strong light adaptation reduced the depolarizing phases more than the hyperpolarizing phase, so that following an adapting stimulus the cell responded to illumination with a pure hyperpolarization (isolated hyperpolarization).4. Removal of external Na(+) ions greatly reduced the initial depolarization. In Na(+)-free sea water the cell responds to dim flashes with a slow depolarization (isolated tail) that involves a conductance decrease, and has the same reversal potential as the hyperpolarizing response recorded from light adapted cells.5. The amplitude of the isolated hyperpolarization and tail varied inversely with the external K(+) concentration.6. It is concluded that in Hermissenda photoreceptors light initiates processes that result in three distinct permeability changes. Following a brief flash there is: a rapid and transient increase in Na(+) permeability that is responsible for the initial depolarization, a less rapid increase in K(+) permeability that is responsible for the hyperpolarizing phase, and a delayed decrease in K(+) permeability that gives rise to the depolarizing tail.

Adaptation, Physiological

Responses of hair cells in the statocyst of Hermissenda.

Responses to mechanical stimulation were recorded from hair cells in the statocyst of Hermissenda crassicornis. The response to a brief stimulus is a depolarizing wave which reaches peak in about 25 msec and decays slowly. 2. Hyperpolarization by extrinsic currents increases the amplitude of the response; depolarization decreases it and eventually reverses its polarity. It is inferred from these results that the primary outcome of the transduction process is an increase of membrane conductance and that the voltage change (generator potential) follows as a secondary event. 3. The features of the conductance change were reconstructed from the time course of the generator potential and the passive properties of the membrane. It was found that the increase of membrane conductance develops slowly and is roughly proportional to the energy delivered by the stimulus. 4. The time course of the conductance change required to reproduce the generator potential is similar to the output of a model involving a sequence of transformations. 5. The generator potential is sensitive to temperature, becoming faster as temperature is raised. This effect is reproduced by the model if the transition rates are assumed to be temperature-dependent, with a Q10 of about 2. 6. It is concluded that a chain of temperature-sensitive processes is interposed between the stimulus and the increase of membrane conductance.

Animals