Abstract. When purple-membrane fragments from Halobacterium halobium are added to one aqueous phase of a positively-charged black lipid membrane, the membrane becomes photoelectrically active. Under normal conditions the steady-state photo-current is extremely low, but increases considerably when the lipid bilayer is doped with proton-permeable gramicidin channels or with a lipophilic acid-base system. These findings indicate that the purple-membrane sheets are bound to the surface of the bilayer, forming a sandwich-like structure. The time-behaviour of the photocurrent may be interpreted on the basis of a simple equivalent circuit which contains the conductance and capacitance of the purple membrane in series with the conductance and capacitance of the lipid bilayer. From the dependence of the photocurrent on the polarization of the exciting light the average angle between the transition moment of the retinal chromophore and the plane of the bilayer was calculated to be about 28 degrees. Furthermore, it was shown that chromophore-free apomembrane binds to the lipid bilayer and that its photoelectrical activity can be restored in situ by adding all-trans-retinal to the aqueous phase.

According to a recent hypothesis the melanin granules in the retinal pigment epithelium of mammals originate from photosensory membrane degradation. To test this hypothesis the retinal pigment epithelium of cattle was kept in tissue culture and exposed to gold-labelled rod outer segments. Gold granules were later detected inside phagosomes, melanosomes and mature melanin granules. Tyrosinase, the key enzyme in melanogenesis, was additionally localized inside phagosomes. These results indicate that in cultured retinal pigment epithelium the matrix of the melanosome can originate from phagosomes. Therefore, the melanosome is a specialized lysosome.

No abstract

Light-activated single channel currents were measured in Limulus ventral photoreceptors in the cellattached configuration at 14°C. The results show three channel types with conductances of 6.2, 10.4 and 28.7 pS. The most active channels have the 10 pS conductance; the open time histograms of these channels could be best fitted by the sum of two exponentials with time constants (and weights) of 0.58 ms (0.78) and 4.32 ms (0.22), suggesting two populations of channels or two open states. The mean open time was 1.38 ms.The open time histogram of the channels with the 29 pS conductance could be best fitted by a single exponential with a time constant of 3.35 ms. First latencies of the 10 pS channels were between 40 and 280 ms but those of the 29 pS conductance channels were > 300 ms. These findings suggest that the two channel types are gated by two different intracellular transmitters or mechanisms.

The light-induced membrane voltage response (receptor potential, ReP) and the absorption change of the intracellularly injected calcium indicator arsenazo III (arsenazo response) were recorded simultaneously in Limulus ventral nerve photoreceptor cells. A double pulse technique was applied for stimulation. After pressure injection of the indicator into the cell absorption changes were measured at 646 nm to obtain a measure of the changes of the intracellular calcium ion concentration. 1. The size of the arsenazo response increases with increasing intensity of the light stimulus. The intensity dependence of the size of the arsenazo response delta A max shows almost no correlate with the peak amplitude of the ReP, but correlates rather well with the time integral of the ReP. 2. Decreasing light adaptation (caused by prolongation of the repetition time of the pulse pairs) leads to an increase in size of the arsenazo response. Also here delta A max correlates better with the time integral of the ReP than with its peak amplitude. 3. Lowering the calcium concentration in the superfusate (from 10 mmol/l to ca. 40 mumol/l) causes after ca. 10 min a drastical diminution of the arsenazo response to values below the noise level, and a less marked reduction in size of the ReP. The falling phase of the ReP is slower. After return to normal calcium concentration the arsenazo response recovers partly in ca. 50 min, while the ReP recovers faster. The results show two opposite correlation between ReP and arsenazo response: Increase in size and duration of the ReP causes a greater transient increase of the intracellular calcium ion concentration. This in turn tends to reduce and shorten the ReP. Which effect dominates obviously depends on the conditions of the experiment: when the calcium concentration in the superfusate is reduced to ca. 40 mumol/l a slow decrease of the ReP is accompanied by a small increase of the intracellular calcium ion concentration.