PubMed Health⌕ Search

Biomedical subjects

P L Walne

Publications and source records attributed to P L Walne.

18 recordsLinked to original sources

The photoreceptor protein of Euglena gracilis.

We isolated the photoactive protein Erh, isolated from the photoreceptor of the unicellular photosynthetic flagellate Euglena gracilis. It is a 27 kDa protein with a photocycle resembling that of sensory rhodopsin, but with at least one stable intermediate. We recorded the absorption spectrum of the parent form of this protein both under native form and in the presence of hydroxylamine and sodium borohydride, and the fluorescence spectra of both the parent and intermediate forms. We suggest that Erh is a rhodopsin-like protein and propose a simple photocycle. This protein shows optical bistability, without thermal deactivation.

Animals↗

In vivo photocycle of the Euglena gracilis photoreceptor.

We present the light-induced photocycle of the paraflagellar swelling of Euglena gracilis. The kinetics of this process was reconstructed by sampling its fluorescence emission and switching the excitation light from 365 nm to 436 nm. Stable intermediates in the photocycle were manifested. The measured millisecond resolution kinetics best fits a Michaelis-Menten equation. The data provide strong evidence that the paraflagellar swelling, a three-dimensional natural crystal of a light-detecting protein, is the true Euglena photoreceptor.

Animals↗

Effects of hydroxylamine, digitonin and triton X-100 on photoreceptor (paraflagellar swelling) and photoreception of Euglena gracilis.

We present experiments that test the effects of agents commonly used in visual pigment investigations, namely hydroxylamine (NH2OH), digitonin and triton X-100, on the photoreceptor and photoreception of Euglena. Hydroxylamine reacts with free and opsin-bound retinal, in aqueous solution, to form stable oximes, whereas digitonin and triton X-100 are the most common extractants of rhodopsin. Since previous data indicate that the chromophore present in Euglena photoreceptor is retinal, we investigated the influence of these chemicals on this organelle. The effects of these agents were studied by means of phase contrast, fluorescence and transmission electron microscopy and photobehaviour experiments. Hydroxylamine inhibited the formation of the Euglena photoreceptor. Photoaccumulation experiments on hydroxylamine-treated cells showed that they are unable to perceive light. Digitonin solubilized the crystalline structure of the photoreceptor, whereas the triton effect was limited to the membranous structures of the cell, leaving the photoreceptor unimpaired.

Animals↗

Effect of pronase treatment on step-down and step-up photophobic responses in Euglena gracilis.

Pronase-treated cells of Euglena gracilis Z show no discernible ultrastructural effects on the photoreceptor apparatus; however, there are physiological effects on swimming speed and on step-up and step-down photophobic responses, especially the latter. Pronase acts differently on the two photophobic responses: the step-down response is completely inhibited after 2 hr., whereas inhibition of the step-up response occurs in only 50% of the cells even after 24 hr. The effects are fully reversible, with step-up recovery quite rapid and step-down recovery considerably slower.

Animals↗

Algal photosensory apparatus probably represent multiple parallel evolutions.

We postulate that algal photoresponse mechanisms are of relatively recent origin and represent numerous parallel evolutions. Functional differences among them are evident, and it is unlikely that any one can be taken as a "model system" representing all. It is probable that the light antenna is the only truly novel part of the apparatus in most cases, with the signal-processing and motile elements being borrowed from some other, more ancient, sensory system. It is thus anticipated that the light-antennae will show the greatest phylogenetic variation, whereas the signal-processors and motile elements will resemble those of other sensory systems in the same groups of organisms.

Biological Evolution↗

Ultrastructure and physical characteristics of proplastids and chloroplasts isolated by zonal centrifugation from Euglena gracilis var. bacillaris.

Proplastids at various stages of development and mature chloroplasts were isolated by zonal centrifugation in a double-sigmoid, sucrose-sorbitol gradient from Euglena gracilis var. bacillaris grown in darkness and subjected to various periods of illumination with white light. Computer programs were used in characterizing physical properties of the isolated organelles and in predicting the location of plastids of different developmental stages in the gradient. The technique permits computation of these parameters in any gradient material. The convoluted nature of the early developing plastids, revealed by electron microscopy, suggests that the frictional coefficients should be taken into account in describing the physical constants of the organelles, since their sedimentation coefficients are lower than would be expected for spherical particles of the same size and density.

Animals↗

Studies on scale morphogenesis in the Golgi apparatus of Pyramimonas tetrarhynchus (Prasinophyceae).

Ultrastructural investigations of scale formation in the cisternae of the Golgi apparatus have been carried out on the prasinophycean flagellate Pyramimonas tetrarhynchus, whose cell surfaces are covered with 6 different scale types, 3 on the flagella and 3 on the cell body. Our results suggest that at least 4 and probably all 6 scale types can be formed together within the same cisterna and that there is some degree of intracisternal differentiation, since the formation of 2 scale types (the small underlayer scales on cell body and flagella) is restricted to the cisternal peripheries, whereas the remaining scale types are formed in the more central portions. Detailed studies of morphogenesis of the larger body scales reveal the earliest identifiable stages as 8-armed figures, with 8 thin arms in the intermediate body scales (IBS), and 4 thick and 4 thin arms in the outer body scales (OBS). From these incipients structures that bear little resemblance to the finished products, the complex, 3-dimensional mature body scales are elaborated in each of the cell's 4 distyosomes, and maintain throughout their different developmental sequences a close relationship to the inner surfaces of the cisternal membranes, as well as a particular orientation within the dictyosomes. Preliminary calculations of total numbers of scales that cover cell and flagellar surfaces are included. The small, underlayer scales, which on the flagella are shown to be arranged in 24 rows, number about 350000; larger scales of more complex construction number about 20 000.

Animals↗

Aliphatic Chains of Esterified Lipids in Isolated Eyespots of Euglena gracilis var. bacillaris.

Isolated eyespot granules of Euglena gracilis Klebs var. bacillaris Pringsheim contained approximately 6% lipids (based on protein). Separation of the lipid extracts by thin layer chromatography revealed four major fractions: wax esters, triacylglycerols, free fatty acids, and phospholipids. Methanolysis of each fraction yielded between 27 and 29 different fatty acids ranging from 12:0 to 22:6. Acetates of the fatty alcohols of the wax fraction consisted of 11:0 to 18:0 carbon chains, with 14:0 being the major component; unsaturated alcohols were not detected.

Journal Article↗

Large Scale Isolation and Purification of Eyespot Granules from Euglena gracilis var. bacillaris.

Large volumes of eyespot granules were isolated from homogenates of Euglena gracilis Klebs var. bacillaris Pringsheim by flotation centrifugation in a Beckman Ti-15 zonal rotor, and were further purified by centrifugation in a swinging bucket rotor. Examination with the electron microscope showed the eyespot granules to be free from other cellular material. Freezing had no apparent effect on the structure or on the absorption properties of the eyespots. Absorption spectra of pure fractions of eyespot granules free of chloroplast contamination showed the previously reported curves in the range of 360 to 520 nanometers, as well as a peak at 660 to 675 nanometers. The procedure for the large scale isolation of eyespot granules from Euglena gracilis is compared with other methods which have employed conventional centrifugation, and the significance of the use of zonal rotors for isolating large quantities of pure eyespot granules is discussed.

Journal Article↗

Photodestruction of chloroplast ultrastructure by red light: location of chlorophyll.

A lethal dose (1 Mrad) of gamma rays was given to wheat leaf tissue to restrict migration of the energy of red light photoexcitation to the photosynthetic pigment system. This dose of gamma radiation had no apparent effect on chloroplast ultrastructure in tissue that received no illumination. Subsequent illumination with red light, used to induce chlorophyll-sensitized photodestruction of ultrastructure, permitted inferences about the location of chlorophyll within the chloroplast. The red light produced extensive disruption in chloroplasts but not in any other cellular organelles. The sequence of events in this photodestruction was different from that which occurs when chloroplasts are destroyed by methods that do not involve illumination. The red light caused peculiar abnormalities only in the internal lamellar system of the chloroplast and not in the envelope or any other regions. These findings support other studies which suggest that chlorophyll is located exclusively in grana and stroma lamellae.

Journal Article↗

Nonphotosynthetic retardation of chloroplast senescence by light.

Excised apical portions of green wheat leaf sections were treated with aminotriazole to prevent formation of new chloroplasts. Illumination retarded the decline in chlorophyll content per leaf section, the disintegration of chloroplast ultrastructure, and the loss of capacity for photosynthetic carbon fixation. We interpret these 3 effects of illumination as facets of a single light effect in retarding chloroplast senescence. This light effect in retarding chloroplast senescence has features differing from characteristics of photosynthetic carbon fixation. For example, A) application of the photosynthetic inhibitor 3-(3,4-dichlorophenyl)-1, 1-dimethylurea did not decrease, and may have even slightly increased, the effectiveness of light; B) although the action spectrum contains peaks in the blue and red regions, it differs from the action spectrum for photosynthetic CO(2) assimilation in wheat; C) in nonphotosynthesizing tissue, application of sugars did not retard chloroplast senescence; D) light saturation was achieved by only a few hundred microwatts/cm(2). Considered together with the well-known light requirement for chloroplast formation, our results indicate that light has a dual, photomorphogenetic control in maintaining the green status of the plant by also exerting a second effect: retarding of senescence of chloroplasts already present.

Journal Article↗