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Biomedical subjects

W L Pak

Publications and source records attributed to W L Pak.

At least 55 records · Page 3Linked to original sources

Drosophila locus with gene-dosage effects on rhodopsin.

Mutations that decrease the amplitude of the prolonged depolarizing afterpotential (PDA) in Drosophila melanogaster have been shown to have reduced rhodopsin content in the rhabdomeres of photoreceptor cells. In the present study, a genetic analysis of a class of third chromosome PDA-defective mutants localized the ninaE locus to the salivary band region 92A-B. In flies with only one copy of this region instead of the normal two copies, and in ninaE heterozygotes, the rhodopsin content of the major class of photoreceptors is reduced. Three doses of this region increase the rhodopsin content of these photoreceptors. These characteristics of the ninaE locus are expected of the structural gene encoding the major species of opsin in the Drosophila compound eye.

Journal Article↗

Light-induced modification of Drosophila retinal polypeptides in vivo.

The effect of light on the polypeptide map profile of the Drosophila eye preparation was examined by two-dimensional polyacrylamide gel electrophoresis. The results show (i) that illuminating the living fly reversibly changes the isoelectric points of three classes of polypeptides specific for the photoreceptor layer and (ii) that the norpA mutation, which prevents the generation of the receptor potential, blocks the modifications.

Animals↗

Freeze-fracture study of the Drosophila photoreceptor membrane: mutations affecting membrane particle density.

The photoreceptor membrane of Drosophila melanogaster (wild type, vitamin A-deprived wild type, and the mutants ninaAP228, ninaBP315, and oraJK84) was studied by freeze-fracture electron microscopy. The three mutations caused a decrease in the number of particles on the protoplasmic face of the rhabdomeric membrane. The ninaAP228 mutation affected only the peripheral photoreceptors (R1-6), while the ninaBP315 mutation affected both the peripheral (R1-6) and the central photoreceptors (R7). The oraJK84 mutation, which essentially eliminates R1-6 rhabdomeres, was found to drastically deplete the membrane particles in the vestigial R1-6 rhabdomeres but not in the normal rhabdomeres of R7 photoreceptors, suggesting that the failure of the oraJK84 mutant to form normal R1-6 rhabdomeres may be due to a defect in a major R1-6 photoreceptor-specific protein in the mutant. In all cases in which both the rhabdomeric particle density and rhodopsin content were studied, the mutations or vitamin A deprivation was found to reduce both these quantities, supporting the idea that at least the majority of the rhabdomeric membrane particles are closely associated with rhodopsin. Vitamin A deprivation and the mutations also reduced the number of particles in the plasma membrane as in the rhabdomeric membrane, suggesting that both classes of membrane contain rhodopsin.

Animals↗

Light-induced pigment granule migration in the retinular cells of Drosophila melanogaster. Comparison of wild type with ERG-defective mutants.

The dependence of pigment granule migration (PGM) upon the receptor potential was examined using several strains of electroretinogram (ERG)-defective mutants of Drosophila melanogaster. The mutants that have a defective lamina component but a normal receptor component of the ERG (no on-transient A [nonA] and tan) exhibited normal pigment granule migration. The mutants that have very small or no receptor potentials (certain no receptor potential A [norpA] alleles), on the other hand, exhibited no PGM. In the case of the temperature-sensitive norpA mutant, norpAH52, normal PGM was present at 17 degrees but not at 32 degrees C or above, corresponding to its electrophysiological phenotype. In the transient receptor potential (trp) mutant, whose receptor potential decays to the baseline within a few seconds during a sustained light stimulus, the pigment granules initially moved close to the rhabdomere when light was turned on but moved away after about 5 s during a sustained light stimulus. All these results lend strong support to the notion that PGM is initiated by a light-evoked depolarization of the receptor membrane, i.e., the receptor potential. However, under certain experimental conditions, the receptor potentials failed to induce PGM in the trp mutant. The depolarization of the receptor, thus, appears to be closely associated with PGM but is not a sufficient condition for PGM.

Animals↗

Mutation that selectively affects rhodopsin concentration in the peripheral photoreceptors of Drosophila melanogaster.

A Drosophila mutant (ninaAP228) that is low in rhodopsin concentration but identical to the wild-type fly in photoreceptor morphology has been isolated. R1-6 photoreceptors of the mutant differ from those of wild type in that (a) the prolonged depolarizing afterpotential (PDA) is absent, (b) concentrations of rhodopsin and opsin are substantially reduced, and (c) intramembrane particle density in the membranes of the rhabdomeres is low. Each of these traits is mimicked by depriving wild-type flies of vitamin A. The ninaAP228 mutation differs from vitamin A deprivation in that in the mutant (a) the rhabdomeric membrane particle density is reduced only in the R1-6 photoreceptors and not in R7 or R8, (b) the PDA can be elicited from the R7 photoreceptors, and (c) photoconversion of R1-6 rhodopsin to metarhodopsin by ultraviolet (UV) light is considerably more efficient than in vitamin A-deprived flies. The absorption properties of the mutant rhodopsin in the R1-6 photoreceptors appear to be identical to those of wild type as judged from rhodopsin difference spectra. The results suggest that the mutation affects the opsin, rather than the chromophore, component of rhodopsin molecules in the R1-6 photoreceptors. The interaction between the chromophore and R1-6 opsin, however, appears to be normal.

Animals↗

Heterogenic components of a fast electrical potential in Drosophila compound eye and their relation to visual pigment photoconversion.

The electroretinogram of the dipteran compound eye in response to an intense flash contains an early, diphasic potential that has been termed the M potential. Both phases of the M potential arise from the photostimulation of metarhodopsin. The early, corneal-negative component, the M1, can be recorded intracellularly in the photoreceptors and has properties similar to the classical early receptor potential (ERP). The M1 is resistant to cold, anaesthesia, and anoxia and has no detectable latency. It depends on flash intensity and metarhodopsin fraction in the manner predicted for a closed, two-state pigment system, and its saturation is shown to correspond to the establishment of a photoequilibrium in the visual pigment. On the other hand, the dominant, corneal-positive component, the M2, does not behave like an ERP. It arises, not in the photoreceptors, but deeper in the retina at the level of the lamina, and resembles the on-transient of the electroretinogram in its reversal depth and sensitivity to cooling or CO2. The on-transient, which is present over a much wider range of stimulus intensity than the M potential, has been shown to arise from neurons in the lamina ganglionaris. Visual mutants in which the on-transient is absent or late are also defective in the M2. It is proposed that the M2 and the on-transient arise from the same or similar groups of second-order neurons, and that the M2 is a fast laminar response to the depolarizing M1 in the photoreceptors, just as the on-transient is a fast laminar response to the depolarizing late receptor potential. Unlike the M1, the M2 is not generally proportional to the amount of metarhodopsin photoconverted, and the M2 amplitude is influenced by factors, such as a steady depolarization of the photoreceptor, which do not affect the M1.

Action Potentials↗

Ionic mechanism for the generation of horizontal cell potentials in isolated axolotl retina.

The ionic mechanism of horizontal cell potentials was investigated in the isolated retina of the axolotl Ambystoma mexicanum. The membrane potentials of both receptors and horizontal cells were recorded intracellularly while the ionic composition of the medium flowing over the receptor side of the retina was changed. The membrane potential of the horizontal cell is highly depender side of the retina was changed. The membrane potential of the horizontal cell is highly dependent on the extracellular concentration of sodium. When the external ion concentration of either chloride or potassium was changed independently of the other, there were shifts in the membrane potential of the horizontal cell which could not be explained by changes in the equilibrium potential of these ions. If the external concentrations of both potassium and chloride ions were varied so that the product of their external concentrations did not change, the shift in the membrane potential of the horizontal cell was in the direction predicted by the Nernst equation. The results are consistent with the suggestion that in the dark the receptors release a synaptic transmitter which increases primarily the sodium conductance of the horizontal cell postsynaptic membrane.

Ambystoma↗

Light-induced voltage noise in the photoreceptor of Drosophila melanogaster.

The Drosophila photoreceptor potential is thought to be composed of discrete unit potentials called bumps. The steady-state receptor potential and the accompanying voltage fluctuations were recorded intracellularly under steady illumination. The occurrence rate, effective amplitude, and duration of the bumps were deduced by assuming a shot noise model. Over a wide range of light intensity, the duration of bumps remained essentially constant (25-30 ms). Below the saturation intensity for the receptor potential, the bump rate was roughly proportional to the intensity, and the adjustment of bumps to smaller size at higher intensity was mainly responsible for the nonlinear behavior of the receptor potential. The reduction in size of bumps at increasing light intensity was found to be due mainly to the diminishing magnitude of the bump current, and not to some other secondary effects. The bump rate saturated at about 3 x 105-106 events/s.

Animals↗

Horizontal cell potentials: dependence on external sodium ion concentration.

The membrane potential of the horizontal cell of the axolotl is highly dependent on the extracellular concentration of sodium. Experimental results reported here are consistent with the suggestion that in the dark the receptors release a synaptic transmitter which increases primarily the sodium conductance of the postsynaptic membrane. Externally applied aspartate or glutamate depolarizes the horizontal cell membrane and eliminates the light response of the horizontal cell. However, it appears to increase the conductances of the postsynaptic membrane to several ions rather than just to sodium ions.

Ambystoma↗

Quantal basis of photoreceptor spectral sensitivity of Drosophila melanogaster.

Small potential fluctuations ("bumps"), boyh spontaneous and light induced, can be recorded intracellularly from the photoreceptors of Drosophila melanogaster. Statistical analyses of these bumps in the spectral range, 400-600 nm, lead to the following interpretations; (a) For weak stimuli at least, these bumps are the quantal units of the receptor potential. (b) Quanta of various wavelengths, when effectively absorbed, will elicit bumps of the same average size. (c) The spectral sensitivity of the receptor potential appears to have its origin in the relative efficiency of quantum bump production at different wavelengths, and not in the intrinsic difference in the properties of bumps produced by quanta of differenct wavelengths.

Animals↗