Effects of melatonin and light on porphyrin synthesis in the bovine retina, pigment epithelium and choroid.

The quantity of porphyrin synthesized in the presence of 10(-3) M delta-aminolevulinic acid (ALA) is several times higher in the bovine pigment epithelium than in the retina. Synthesis in the retina was found to be increased by illumination, whereas synthesis in the pigment epithelium was decreased if the whole anatomical unit (retina-pigment epithelium-choroid) was cultivated together. The quantity of porphyrin synthesized in the presence of 10(-3) M ALA or 10(-6) M melatonin was different when the pigment epithelium and retina were separated. The combination 10(-3) M ALA with 10(-6) M melatonin inhibited retinal porphyrin synthesis after green light adaptation, while in the pigment epithelium green light adaptation induced porphyrin synthesis. It is postulated that the light-sensitive porphyrin-haeme synthesis of the retina-pigment epithelium-choroid functional unit may serve and modulate the synthesis of guanylate cyclase for cGMP.     

Inhibitory effects of dibutyryl cyclic AMP and theophylline on the melanosome transformation in the embryonic chick pigmented retina cultured in vitro.

When the retinal pigment epithelial cells of chick embryo are cultured in monolayer conditions, the pigment granules are lost from the cytoplasm. The first structural change in depigmentation is the transformation of pigment granules into the degradative organelles designated as the dense body and melanosome complex. The cells are grown in medium containing DBcAMP of various doses from 10^?5 to 10^?2M. Cell proliferation is retarded by treatment with DBcAMP (10^?3M). The transformation of pigment granules is almost completely prevented in all 1-day cultured cells. In 5-day cultured cells continuously treated with more than 10^?4M, the transformation is not only prevented, but the synthesis of pigment granules is stimulated. A similar result is obtained by the administration of 10^?3M theophylline. 5′-AMP does not prevent the transformation of pigment granules but seems to stimulate the synthesis of pigment granules. On the other hand, cGMP is ineffective both on prevention of transformation and on synthesis of pigment granules. The mechanisms of the transformation of pigment granules are discussed.     

Cloning and regulation of Erwinia herbicola pigment genes.

The genes coding for yellow pigment production in Erwinia herbicola Eho10 (ATCC 39368) were cloned and localized to a 12.4-kilobase (kb) chromosomal fragment. A 2.3-kb AvaI deletion in the cloned fragment resulted in the production of a pink-yellow pigment, a possible precursor of the yellow pigment. Production of yellow pigment in both E. herbicola Eho10 and pigmented Escherichia coli clones was inhibited by glucose. When the pigment genes were transformed into a cya (adenylate cyclase) E. coli mutant, no expression was observed unless exogenous cyclic AMP was provided, which suggests that cyclic AMP is involved in the regulation of pigment gene expression. In E. coli minicells, the 12.4-kb fragment specified the synthesis of at least seven polypeptides. The 2.3-kb AvaI deletion resulted in the loss of a 37K polypeptide and the appearance of a polypeptide of 40 kilodaltons (40K polypeptide). The synthesis of the 37K polypeptide, which appears to be required for yellow pigment production, was not repressed by the presence of glucose in the culture medium, as was the synthesis of other polypeptides specified by the 12.4-kb fragment, suggesting that there are at least two types of gene regulation involved in yellow pigment synthesis. DNA hybridization studies indicated that different yellow pigment genes exist among different E. herbicola strains. None of six pigmented plant pathogenic bacteria examined, Agrobacterium tumefaciens C58, Cornyebacterium flaccumfaciens 1D2, Erwinia rubrifaciens 6D364, Pseudomonas syringae ATCC 19310, Xanthomonas campestris 25D11, and "Xanthomonas oryzae" 17D54, exhibited homology with the cloned pigment genes.     

Terminal Synthesis of Xanthommatin in DROSOPHILA MELANOGASTER. III. Mutational Pleiotropy and Pigment Granule Association of Phenoxazinone Synthetase

Phenoxazinone synthetase, which catalyzes the condensation of 3-hydroxykynurenine to xanthommatin, the brown eye pigment of Drosophila, is shown to exist in association with a particle which resembles the cytologically defined Type I pigment granule. Several classical eye color mutants (v, cn, st, ltd, cd, w), including two which effect other enzymes in the xanthommatin pathway (v, cn), have low levels of phenoxazinone synthetase activity and disrupt the normal association of the enzyme with the pigment granule. A model is proposed depicting several structural and enzymatic interrelationships involved in the developmental control of xanthommatin synthesis in Drosophila.     

Thylakoid membrane biogenesis in Chlamydomonas reinhardtii 137+. II. Cell-cycle variations in the synthesis and assembly of pigment

Synthesis of the chlorophyll and the major carotenoid pigments and their assembly into thylakoid membrane have been studied throughout the 12-h light/12-h dark vegetative cell cycle of synchronous Chlamydomonas reinhardtii 137+ (wild-type). Pulse exposure of cells to radioactive acetate under conditions in which labeling accurately reflects lipogenesis, followed by cellular fractionation to purify thylakoid membrane, allowed direct analysis of the pigment synthesis and assembly attendant to thylakoid biogenesis. All pigments are synthesized and assembled into thylakoids continuously, but differentially, with respect to cell-cycle time. Highest synthesis and assembly rates are confined to the photoperiod (mid-to-late G1) and support chlorophyll and carotenoid accretion before M-phase. The lower levels at which these processes take place during the dark period (S, M, and early-to- mid G1) have been ascribed to pigment turnover. Within this general periodic pattern, pigment synthesis and assembly occur in a "multi- step" manner, i.e., by a temporally-ordered, stepwise integration of the various pigments into the thylakoid membrane matrix. The cell-cycle kinetics of pigment assembly at the subcellular level mirror the kinetics of pigment synthesis at the cellular level, indicating that pigment synthesis not only provides chlorophyll and carotenoid for thylakoid biogenesis but may also serve as a critical rate-determinant to pigment assembly.     

A 4-vinylprotochlorophyllide complex accumulated by "phofil" mutant of Rhodopseudomonas spheroides. An authentic intermediate in the development of the photosynthetic apparatus.

A photosynthetically competent mutant strain of Rhodopseudomonas spheroides was isolated. In addition to bacteriochlorophyll, this organism produced particle-bound precursor 4-vinylprotochlorophyllide. The spectral characteristics of the pigment complexes(es) accumulated in the culture medium were very variable. The spectral form occurring within the bacteria was characterized from fluorescence data. Its particle weight, 130 000, was determined by Sephadex G200 filtration. The main components of the complex were protein, lipid and pigment (6.8:61, w/w). As indicated by qualitative analysis, the lipid components were characteristic constituents of the photosynthetic membrane. Kinetics of pigments synthesis showed that the total pigment synthesis was not affected by the mutation; bacteriochlorophyll content was always lower in the mutant than in the parent strain. The repigmentation process was followed by fluorescence emission. The results indicated that the mutation affected membrane component synthesis required for the bacteriochlorophyll(ide) incorporation. The pigment complex was concluded to be an authentic intermediate in photosynthetic apparatus morphogenesis. The reasons for its excretion are discussed.     

Red pigment in Bacillus megaterium spores.

Bacillus megaterium QM B1551 spores contained a unique red pigment in their membranes that was not found in other species. This red pigment, presumably a carotenoid, was synthesized about the time of dipicolinic acid synthesis during sporulation and was associated with the forespores. A yellow pigment was synthesized during sporulation in rich medium and was found in the mother cell compartment. Although the yellow pigment was also associated with spores, it could be removed by two different extraction procedures without impairing germination; it was absent when sporulation occurred in a minimal medium. Although the yellow pigment of the mother cell appeared to be dispensable, the red pigment may serve a more critical function, such as membrane stabilization.     

Effect of histidine on purine nucleotide synthesis and utilization in Neurospora crassa.

Histidine affects de novo purine nucleotide synthesis and purine nucleotide pool utilization in Neurospora crassa. The former effect was assessed qualitatively by the presence or absence of purple pigment production in ad3B and ad3A mutants. Tryptophan also affected the de novo purine nucleotide synthesis. The effect of histidine on purine nucleotide pool utilization resulted in stimulated germination of ad8 and ad4 mutant conidia in adenine-deficient medium. Increased germination was correlated with increased net levels of nucleic acids in these strains. Possible mechanisms for the dual action of histidine are discussed.     

Negative effect of ammonium nitrate as nitrogen source on the production of water-soluble red pigments by Monascus sp.

Ammonium salts, especially ammonium nitrate, have been used as nitrogen sources for production of traditional water-insoluble Monascus pigments. However, we noted that defined media employing NH₄NO₃ as the sole nitrogen source in fermentations supported only poor pigment production by Monascus sp., and the pigments produced were mainly cell-bound. NH₄NO₃ was found not to (a) repress pigment synthase formation, (b) enhance synthase decay, or (c) serve as a nitrogen source for pigment production by resting cells; it had a weak inhibitory effect on the action of pigment synthase(s). The high level of cell-bound did not exert a feedback effect on the further synthesis of pigments. These observations indicate that the reason why NH₄NO₃ supports only low pigment production during fermentations is the poor ability of NH₄NO₃ to donate nitrogen in the Schiff-base reaction converting orange pigments to red ones.     

Studies in Carotenogenesis. Some Observations on Carotenoid Synthesis in Two Varieties of Euglena gracilis.

Euglena gracilis v. bacillaris and E. gracilis v. fuscopunctata produce the same three carotenoids, β-carotene, lutein and neoxanthin in approximately the same relative amounts. Lutein is the major pigment and β-carotene represents about 10–15% of the total. The concentration of carotenoids in E. gracilis v. bacillaris is very high, reaching 700 mg./100 g. dry weight.
Streptomycin (0.02%, w/v) and darkness reduced growth of E. gracilis v. bacillaris by about 50% and carotenoid synthesis by about twenty times. Diphenylamine (1/70,000) reduced growth and carotenoid synthesis equally (about 50%). In no case was the synthesis of more saturated polyenes (the phytofluene series) stimulated.
The nature of the eye spot pigment is discussed.     

Studies on the biosynthesis of aspergillin by Aspergillus niger.

Two inhibitors of the biosynthesis of aspergillin, the black spore pigment of Aspergillus niger, have been investigated. 2,4-Dithiopyrimidine exerted its inhibitory effect by intracellularly chelating cupric ion required for normal pigmentation. Dimethylsulfoxide prevented the synthesis of certain phenolic precursors of the native pigment. Partial purification and characterization of pigments from mature cultures revealed the presence of at least three components: (i) a high-molecular-weight (approximately 20,000) native pigment fraction in untreated mold cultures, (ii) a lower-molecular-weight (approximately 5,000) melanin pigment found in both types of inhibited cultures, and (iii) a low-molecular-weight (368) green pigment found only in the 2,4-dithiopyrimidine-inhibited cultures and proposed to be a pentacyclic quinonoid derivative. A pathway for aspergillin biosynthesis is suggested based on these results.     

Lichtinduzierte Farbstoffbildung bei Saccharomyces cerevisiae var. ellipsoideus

Zusammenfassung Die lichtinduzierte Pigmentbildung wird in Abhängigkeit von verschiedenen Faktoren untersucht und ihre Kinetik gemessen. Der Farbstoff entsteht nur in einem stickstoffhaltigen Medium. — Der Temperaturbereich, innerhalb dessen Pigmentbildung möglich ist, liegt zwischen 14° und 39° C mit einem Optimum bie 29° C. — Für die Anfangskonzentration von Glucose im Medium wird ein Optimum bei 3% gefunden. Nach Verbrauch der Glucose hört die Farbstoffbildung auf, sofern nicht ein anderer Faktor begrenzend wirkt wie z. B. hohe Lichtintensität, durch die die Pigmentproduktion stark eingeschränkt, schließlich sogar völlig verhindert wird. In Abhängigkeit von der Lichtintensität findet sich ein Optimum der Pigmentbildung je nach Lichtsensibilität der Zellen zwischen 26 und 45 kerg · sec^-1 · cm^-2. Hohe Lichtintensität wirkt zerstörend auf bereits entstandenes Pigment. — Die Aktivität des pigmentbildenden Systems sowie die erreichbare Farbstoffendkonzentration hängen ferner vom Beginn der Belichtung ab. Nennenswerte Pigmentsynthese ist nur möglich bei einem Belichtungsbeginn innerhalb der ersten 10–12 Std und ist um so intensiver, je früher die Belichtung einsetzt. — Bei im übrigen günstigen Kulturbedingungen arbeitet das pigmentbildende System nur während der Lichteinwirkung. — Kinetische Untersuchungen der Farbstoffbildung zeigen, daß das Pigment im Medium entsteht und von dort in die Zellen aufgenommen wird. Demnach muß eine Synthese von Pigmentvorstufen in den Zellen und ihre Abgabe an das Medium angenommen werden.

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Light-induced pigment production by Saccharomyces cerevisiae var. ellipsoideus Physiology of pigment production

Summary Light-induced pigment synthesis by cells of Saccharomyces cerevisiae var. ellipsoideus is investigated under the influence of various factors, and kinetic studies of its formation are performed. Pigmentation only occurs with nitrogen in the medium. The temperature favouring pigment production ranges between 14° C and 39° C, and has its optimum at 29° C. Optimum glucose concentration in the medium is 3%. Pigment formation continues until glucose is completely exhausted provided that no other factor — as for instance high light intensity — limits pigment synthesis before. The lowest possible light intensity for pigment production was found at 3 kerg · sec^-1 · cm^-2, the optimum lies between 26 and 45 kerg ¢ sec^-1 · cm^-2 — depending on the light sensibility of the cells. Still higher light intensities finally destroy part of the pigment already formed. — There is a clear influence of the onset of light exposure a) on the activity of the pigment producing system and b) on the end concentration of pigment. Only cells exposed to light within the first 10–12 hours of culture give rise to considerable pigment synthesis which is the more active the more the onset of light approaches that of the culture. If all other factors are favourable the pigment producing system once working continues to do so only as long as light is on. —Kinetic experiments show the pigment to be formed in the medium and then to be incorporated by the cells. It must be concluded therefore that pigment precursors are synthesised in the cell and from there transferred into the medium.

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Herrn Prof. Dr. W. Simonis zum 60. Geburtstag gewidmet.     

Pigment synthesis in the Himalayan mouse

The effect of temperature on pigment synthesis in adult and juvenile Himalayan mice was investigated. Since pigment synthesis only occurs in actively growing hair, adult mice were plucked to induce hair growth. The extent of darkening of the hair was recorded by photography against a reference scale. The presence of pigment granules in hair follicles was investigated histologically. Housing adult and juvenile mice at 15 degrees C results in the synthesis of pigment in growing hair follicles whereas housing at 30 degrees C results in the absence of pigment granules in the growing hair follicles.     

Radioautographic study of melanin synthesis in cells of the pigmented epithelium of the retina in adult tritons following surgical removal of the retina]

The results are given for the autoradiographic study of melanin synthesis in the pigment epithelium cells of eye retina in the adult newts by the incorporation of 3H-dioxyphenylalanine (3H-DOPA). The synthesis of melanin began 4--8 days after the operation in the peripheral parts of the eye and but it was not observed in the central region of the eye fundus even 25 days alter the operation. A suggestion is put forward to the effect of the existence of different mechanisms for the initiation of melanin synthesis in these subpopulations of pigment epithelium.     

The effect of light on four protochlorophyllide-binding polypeptides of barley (Hordeum vulgare)

Protochlorophyllide-binding proteins were investigated and possible changes in the pigment-protein association during light-induced chlorophyll synthesis were analyzed. Three major results were obtained. (1) Four protochlorophyllide-binding polypeptides were separated electrophoretically on polyacrylamide gels and visualized by their fluorescence. The number and size of these protochlorophyllide-binding proteins isolated from darkgrown and illuminated plants were not affected by light. (2) The association of pigment with these proteins was studied using exogenous [³H]protochlorophyllide as substrate in an NADPH-dependent in vitro chlorophyll synthesis assay. It was found that a constant exchange of protein-bound and free pigment occurs and that freshly synthesized chlorophyllide does not accumulate on any of the four pigment-binding proteins in vitro. (3) NADPH does not affect the amount of pigment bound to protein in vitro, even during chlorophyll synthesis which occurred only in the presence of NADPH.     

Trypsin-induced cell surface changes in ascidian embryonic cells: regulation of differentiation of a tissue-specific protein.

Developing animal quartets removed surgically from 8-cell stage of Ascidia malaca and Phallusia mamillata have been treated for a short time with a low concentration of trypsin. The result is a differentiation of some neural structures, of pigment spots and of a tissue-specific enzyme, brain pigment cell tyrosinase. Tyrosinase activity, as detected histochemically, appeared in the pigment cells some hours before the normal time independently of any inductive interactions with related embryonic tissues. A study with the electron microscope has given evidence of the presence of presumed nervous cells and melanin granules related to them. An autoradiographic study using [³H]uridine has demonstrated presumed RNA synthesis which suggests gene activation. The results are discussed in relation to the possible role of the plasma membrane during embryonic development.     

Melanin granule metabolism in the iris and retinal pigment epithelium in adult tritions after completion of eye regeneration

It was concluded that the newly synthesized melanin granules were replaced in the pigmented tissues of the newt eye on the basis of redistribution of the cells of pigment epithelium of retina and iris labelled by 3H-DOPA 2.5 and 6.5 months after the isotope injection. The replacement of melanin granules and, correspondingly, melanin synthesis proceed more actively in the peripheral zones of the pigment epithelium of retina. The depigmentation of cells preceding the melanin synthesis appears to be realized with the participation of macrophages.     

Chloroplast autonomy in pigment synthesis

Summary An experimental design developed usingAcetabularia permits a novel approach to studies of cytoplasmic genetic functions. The site of the genes for the enzymes of the plastid pigment pathways were examined by 1. determining pigment content per cell and per chloroplast in intact cells and during long periods of enucleate cell growth, 2. comparing pigment synthetic activities of plastids isolated from intact and enucleate cells at various times postenucleation and 3. comparing the ability of intact and enucleate cells to modulate pigment content in response to various light regimens.Intact cells grow and increase their pigment content exponentially. Enucleate cells grow at the control rate for several weeks and increase their chloroplast number and pigment content proportionally. Pigment content per plastid remains constant in both intact and enucleate cells. Pigment synthesis in isolated chloroplasts from enucleate cells is normal up to 65 days post-enucleation when compared with isolated chloroplasts from intact control cells. Enucleate cells differentially modulate their pigment content in response to various light regimens in a manner indistinguishable from normal cells.The problems in interpreting these and other results are discussed and it is concluded that plastid autonomy in pigment synthesis is the simplest explanation.     

FACE条件下大气O3浓度增高对小麦剑叶光合色素含量的影响

应用FACE研究平台,采用烟农19、扬麦16、嘉兴002、扬麦15和扬辐麦2号等5个小麦品种,以O₃自然浓度为对照,研究了大气O₃浓度增高50%对不同小麦品种剑叶光合色素含量的影响.结果表明：开放式大气O₃浓度增高条件下,小麦剑叶叶绿素a (Chl a)、叶绿素b (Chl b)、叶绿素总量Chl(a+b)和类胡萝卜素含量在孕穗期和开花期与对照差异不显著,而花后各时期均不同程度的降低,其中,Chl a、Chl b和Chl(a+b)含量下降达显著水平,说明大气O₃浓度增高对叶绿素合成影响较小,但加速了其衰降过程.不同品种小麦剑叶光合色素含量对大气O₃浓度增高的反应存在基因型差异,扬麦15和嘉兴002对大气O₃浓度增高的敏感性弱于扬麦16、扬辐麦2号和烟农19.在籽粒灌浆盛期(花后21 d左右),剑叶Chl a、Chl b和Chl(a+b)含量与千粒重呈显著正相关.