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Cross-phyletic bioactivity of arthropod neurohormones and molluscan ganglion extracts: evidence of an extended peptide family.

Two structurally related arthropod neuropeptides, red pigment concentrating hormone (RPCH) and adipokinetic hormone (AKH), are potent excitors of the heart of the clam Mercenaria mercenaria. The response is bimodal: whereas the threshold for affected hearts is 1-3 X 10(-9) M, about 40% of the preparations are virtually unresponsive. Aqueous extracts of Mercenaria ganglia contain a substance which concentrates the red pigment in the erythrophores of intact destalked Uca pugilator and even of its isolated legs. This substance is retained on Sephadex G-15 and co-elutes with synthetic shrimp RPCH. The active fractions also concentrate the erythrophores and the leucophores of destalked shrimp (Penaeus). Neither dopamine nor the molluscan neuropeptide FMRFamide had any chromatophorotropic effect in these assays. The activity of the ganglion extracts was abolished by digestion with chymotrypsin. In conclusion, molluscan ganglion extracts contain a peptide factor, possibly an analog of RPCH, that concentrates the pigments of crustacean chromatophores by a direct action on the cells.

Animals↗

Genetic and experimental studies on a new pigment mutant in Xenopus laevis.

White lethal (wl) is a recessive mutation affecting the differentiation of the three types of chromatophores in Xenopus laevis and eventually leading to the death of the mutants around stage 50. Melanophores appear at st. 33 but differentiate abnormally, remaining pale grey, and do not proliferate after st. 41. The rare xanthophores present contain only a few differentiated pterinosomes, and the iridophores consist of noniridescent white dots. When the albino gene (ap) is combined with wl, melanophores do not differentiate. Reciprocal heterotopic and orthotopic trunk neural crest grafts have shown that the defect is intrinsic to the neural crest cells but is not due, in the case of melanophores, to a tyrosinase deficiency as revealed by the dopa reaction. The mode of action of the gene, the abnormal pattern, and lethality are discussed.

Animals↗

Ultrastructural analysis of iridophore organellogenesis in a lizard, Sceloporus graciosus (Reptilia: Phrynosomatidae).

Transmission electron microscopy (TEM) data from the ultrastructure of lizard skin iridophores (reflective dermal chromatophores) are used to illustrate the organellogenesis of small rectangular reflecting platelets, which are the color-generating components of these cells. During the development of reflecting platelets, crystals are deposited within double-membraned vesicles from electron-dense material located within the vesicles. The crystals are initially small but expanded lengthwise eventually to fill the vesicle that contains them. The inner membrane then tightly surrounds the crystal whereas the outer membrane is much more loosely associated with the inner-membrane-bound crystal. These observations allow discussion of the possible origin of the precursor double-membraned vesicles from endoplasmic reticulum (ER) and Golgi-derived vesicles. A model is proposed that incorporates our findings and other published reports to explain the origin of the precursor double-membraned vesicles via three alternative pathways.

Animals↗

Observations on the development of unusual melanization of leopard frog ventral skin.

The ontogeny of ventral pigmentation of two species of leopard frog, Rana pipiens and R. chiricahuensis, was examined by light microscopy and transmission electron microscopy to reveal how the unusual melanistic ventral pigmentation of R. chiricahuensis is achieved at the cellular level. Ventral skin of R. pipiens is always white. Ventral skin of adult R. chiricahuensis is white when frogs are background-adapted to a white substrate, but ventral skin becomes nearly as dark colored as the dorsal skin when frogs darken in response to a black background. Skin samples from tadpoles of both species, newly metamorphosed frogs, and adult frogs were analyzed for chromatophore composition and distribution. Ventral skin of R. pipiens larvae, newly metamorphosed frogs, and adults and of R. chiricahuensis larvae was white due to abundant iridophores and no melanophores. Melanophore density in the ventral integument of R. chiricahuensis was 9.1 +/- 2.8/mm2 in newly metamorphosed frogs and 87.0 +/- 4.8/mm2 in adult frogs. Pigment within ventral melanophores migrated during physiological color change during background adaptation.

Animals↗

Characterization of chromatophorotropic neuropeptides from the kuruma prawn Penaeus japonicus.

Three chromatophorotropic neuropeptide hormones were purified from an aqueous extract of the sinus glands of the kuruma prawn Penaeus japonicus by two steps of reverse-phase HPLC and their amino acid sequences determined. One of them was found to show pigment concentrating activity and to have an amino acid sequence identical with that of the known red pigment concentrating hormone (RPCH), and therefore it was named Pej-RPCH. The other two peptides showed pigment dispersing hormone (PDH) activity and were named Pej-PDH-I and -II. They both consisted of 18 amino acid residues with a free amino-terminus and an amidated carboxyl-terminus, the sequences of Pej-PDH-I and -II being NSELINSLLGIPKVMTDAamide and NSELINSLLGLPKFMIDAamide, respectively. Three amino acid residues at positions 11, 14, and 16 differed between the two PDHs. Pej-PDH-II was about 5-, 7-, and 10-fold more potent than Pej-PDH-I for erythrophores, xanthophores, and melanophores, respectively. The major reason for the difference in potency between the two PDHs was attributed to differences in residues at position 16. In addition, they were found to be produced by a single individual. The order of sensitivity of the four types of chromatophores to Pej-RPCH and both PDHs was found to be erythrophores = xanthophores > melanophores > leukophores.

Amino Acid Sequence↗

Ultrastructure of the extracutaneous pigment cells in the plaice (Pleuronectes platessa, L., Teleostei).

The extracutaneous pigment cell system of the plaice (Pleuronectes platessa L.) was examined by light and electron microscopy in selected regions, including two cutaneous regions for comparison. The extracutaneous pigmentation consists of guanocytes and melanocytes with differing distributions within the body. The eyeless side lacks melanocytes. The pigment cells are differentiated as very flat elements with long processes. They display an affinity for loose connective tissue at boundary layers such as the peritoneal epithelium, organ capsules or blood vessels, to which they are parallelly arranged at a very constant distance. In some locations guanocytes are intimately associated with melanocytes forming "reduced chromatophore units". Extracutaneous pigment cells are poor in mitochondria, endoplasmic reticulum, microfilaments, caveolae intracellulares, ribosomes and glycogen granules, all of which are more abundant in cutaneous pigment cells and pigment cells of the eye. In extracutaneous guanocytes the crystals are loosely arranged parallel to the cell surface, in cutaneous guanocytes perpendicular. Cells with rod-like vesicular cisternae are described as "guanoblasts". No single pigment cell was found exhibiting different types of pigment granules. The varying colors of extracutaneous pigmentation arise from varying combinations of guanocytes and melanocytes in addition to the color of the tissue itself.

Animals↗

Reflector cells in the skin of Octopus dofleini.

The cells that form the reflecting layer beneath the chromatophore organs of the octopus are conspicuous elements of its dermal chromatic system. Each flattened, ellipsoidal reflector cell in this layer bears thousands of peripherally radiating, discoidal, reflecting lamellae. Each lamella consists of a proteinaceous reflecting platelet enveloped by the plasmalemma. The lamellae average 90 nm in thickness and have variable diameters with a maximum of about 1.7 micrometer. Sets of reflecting lamellae are organized into functional units called reflectosomes. The lamellae in each reflectosome form a parallel array - similar to a stack of coins. The average number of lamellae in a reflectosome is 11. Adjacent lamellae are uniformly separated by an extracellular gap of about 60 nm in embedded specimens. The reflectosomes are randomly disposed over the surface of the reflector cell. The observed organization of the reflectosomes is compatible with its role as a quarter-wave thin-film interference device. The alternating reflecting lamellae and intelamellar spaces constitute layers of high and low refractive indices. Using measurements of the thicknesses and refractive indices of the platelets and interlamellar spaces, we have calculated that the color of reflected light should be blue - green, as seen in vivo. The sequence of events leading to the definitive arrangement of the reflectosomes is uncertain. The reflector cells of O. dofleini are compared and contrasted with the iridophores of squid.

Animals↗

Hybrid pigment organelles in an invertebrate.

Observations of a number of vertebrate chromatophores have revealed the presence of more than one type of pigment organelles, suggesting that the different types are all derived from an equipotential organelle able to differentiate into any of the major pigment-containing organelles (Bagnara, 1972). Observations are presented concerning the occurrence of hybrid pigment inclusions, i.e., all kinds of intergrades between melanosomes, pterinosomes, and reflecting platelets in pigment cells of the daddy-long-legs. It therefore seems possible that pigment organelles in some invertebrates may also be derived from a common pluripotential primordial organelle.

Animals↗

The pigmentary system of planaria. II. Physiology and functional morphology.

The pigmentary system of the planaria, Dugesia gonocephala s.l. (Platyhelminthes, Turbellaria, Tricladida), consists of granules contained in chromatophore cells distributed in the parenchyma tissue. The administration of MSH release-inhibiting Factor (M.I.F.) leads to an easily observable general decolouration of the animal due to the migration of the pigment granules towards the deeper-lying cell nucleus. In planarians bisected transversely through the pharyngeal region, the decolouration occurs only in the cephalic segment, and the caudal segment remains dark. When, however, the decapitated caudal segment regenerates a head region, a decolouration response occurs when exposed to M.I.F. The significance of these results is discussed, and an hypothesis on the hormonal regulation of the pigmentary system is proposed.

Animals↗

On the dielectrically observable consequences of the diffusional motions of lipids and proteins in membranes. 2. Experiments with microbial cells, protoplasts and membrane vesicles.

The dielectric properties of suspensions of intact cells of Methylophilus methylotrophus, Paracoccus denitrificans and Bacillus subtilis have been measured in the frequency range 1 kHz to 13 MHz. All possess a pronounced dispersion corresponding in magnitude and relaxation time to the "beta-dispersion" in a terminology defined by Schwan [Adv. Biol. Med. Phys. 5:147-209 (1957)]. The latter two strains, but not M. methylotrophus, also possess a substantial alpha-dispersion. The relaxation time of the beta-dispersion of B. subtilis is significantly lower than that of the other two strains, due to the higher internal K+ content of this Gram-positive organism. Treatment of P. denitrificans or B. subtilis with lysozyme greatly reduces the magnitude of the alpha-dispersion; in the latter case it is virtually abolished. The magnitude of both the alpha- and beta-dispersions of protoplasts of these organisms is significantly decreased by treatment with the cross-linking reagent glutaraldehyde, indicating that diffusional motions of the lipids and/or proteins in the protoplast membranes contribute to the dielectric relaxations observed in this frequency range. Such motions cannot be unrestricted, as in the "fluid mosaic" model, since the relaxation times of the lipids and proteins, if restricted by hydrodynamic forces alone, should then correspond, in protoplasts of this radius (0.4-0.5 micron), to approximately 10 Hz. Even after treatment of the (spherical) protoplasts with glutaraldehyde, the breadth of the remaining beta-dispersion is still significantly greater than (a) that of a pure Debye dispersion and (b) that to be expected solely from a classical Maxwell-Wagner-type mechanism. It is recognised that the surfaces of the protein complexes in such membranes extend significantly beyond the membrane surface as delineated by the phospholipid head-groups; such molecular granularity can in principle account for the broadened dielectric relaxations in the frequency range above 1 kHz, in terms of the impediment to genuinely tangential counterion relaxation caused by the protruding proteins themselves. The relaxation time of a previously observed, novel, low-frequency, glutaraldehyde-sensitive (mu-) dispersion in bacterial chromatophore suspensions, as well as that of their alpha-dispersion, is significantly increased by increasing the aqueous viscosity with glycerol. This finding is consistent with the view that, from a dielectric standpoint, the motions of charged proteins (and lipids) in biological membranes are rather tightly coupled to those of the adjacent ions and dipoles in the electric double layer.(ABSTRACT TRUNCATED AT 400 WORDS)

Bacillus subtilis↗

Resolution and reconstitution of Rhodospirillum rubrum pyridine dinucleotide transhydrogenase. II. Solubilization of the membrane-bound component.

The Rhodospirillum rubrum pyridine dinucleotide transhydrogenase system is comprised of a membrane-bound component and an easily dissociable soluble factor. Active transhydrogenase complex was solubilized by extraction of chromatophores with lysolecithin. The membrane component was also extracted from membranes depleted of soluble factor. The solubilized membrane component reconstituted transhydrogenase activity upon addition of soluble factor. Various other ionic and non-ionic detergents, including Triton X-100, Lubrol WX, deoxycholate, and digitonin, were ineffectual for solubilization and/or inhibited the enzyme at higher concentrations. The solubilized membrane component was significantly less thermal stable than the membrane-bound component. None of the pyridine dinucleotide substrate affected the thermostability of the solubilized membrane-bound component, whereas NADP+ and NADPH afforded protection to membrane-bound component. NADPH stimulated trypsin inactivation of membrane-bound component to a greater extent that NADP+, but inactivation of solubilized membrane component was stimulated to the same extent by both pyridine dinucleotides. The solubilized membrane component appears to have a slightly higher affinity for soluble factor than does the membrane-bound component.

Bacterial Chromatophores↗

A comparison of electron transport and photophosphorylation systems of Rhodopseudomonas capsulata and Rhodospirillum rubrum. Effects of antimycin A and dibromothymoquinone.

The photophosphorylation systems of Rhodopseudomonas capsulata and Rhodospirillum rubrum chromatophores have been compared in respect to the effects of artificial electron carriers [N-methylphenazonium methosulfate (PMS) and diaminodurene], reducing agents (ascorbate in particular), and various quinones in the absence and presence of the electron transport inhibitors antimycin A and dibromothymoquinone (DBMIB). In addition, the effects of both inhibitors on photosynthetic electron transport through cytochromes b and c has been followed. From the results obtained, it appears that in both organisms: a) ubiquinone functions as an electron carrier between the cytochromes, and b) both antimycin A and DBMIB inhibit cyclic electron flow in the segment...cytochrome b leads to ubiquinone leads to cytochrome c..., but at different sites. The systems apparently differ mainly in respect to the nature of the electron flow by-pass "shunt" that is evoked in the presence of PMS; thus, in R. rubrum, PMS catalyzes a shunt that by-passes both cytochrome b and ubiquinone, whereas in Rps. capsulata the PMS shunt seems to circumvent only ubiquinone.

Antimycin A↗

[Biomicroscopic aspects of normal iris tissue in culture (author's transl)].

The cells shown were cultivated from iris pieces of 1-5 mg wet weight incubated in TCM 199 after adding 20% calf serum and antibiotics. Different morphologic aspects allowed some coordination of the cells to the corresponding iris layer. Even after 1 year's incubation, fibroblastic stroma cells showed a radiate growth from the insemination point. Pigment cell-like elements were packed with yellowish, bright, and intensely refracting spherical bodies, indicating a specific function. An analogous finding in the human eye after a perforating iris lesion was growth of chromatophores on the anterior lens capsule.

Animals↗

Evolution of photosystems of photosynthetic organisms.

It is generally accepted that two photosystems function successively in photosynthetic electron transport chain of plants and algae. The interaction of these photosystems results in the enhancement of photosynthesis. It was suggested that only one photosystem is present in purple bacteria, the most primitive photosynthetic organisms. The functioning of this photosystem is accompanied by absorption changes at 890 nm. Recently new spectral changes were found in Chramatium chromatophores under reductive conditions, more favorable for bacterial growth. Some of that spectral changes take place even at liquid nitrogen temperature. It is proposed these absorption changes could be related to other photosystem functioning in low potential region. Such a photosystem is necessary for reduction of NAD in Chromatium, for which the reverse electron transport to NAD was not shown. In contrast to photosystems of plants, the bacterial photosystems appear to function independently because the enhancement of bacterial photosynthesis is not found. Apparently the evolution of photosystems involved interaction between independent photosystmes, one of them functioning under more oxidative conditions.

Bacterial Chromatophores↗

Ultrastructural changes accompanying mitosporogenesis in Ectocarpus parvus.

The plurilocular zoosporangium of Ectocarpus parvus Saunders in initiated as a uniseriate branch from a vegetative cell. Subsequent anticlinal and periclinal divisions of the cells leads to enlargement of the sporangium and the formation of multiple locules. Each locule containes a single biflagellate zoospore. Ultrastructural changes accompanying these events include a reduction in the size of chromatophores, a reduction in the number of dictyosomes and the appearance of plasmalemmasomes in later stages. Enlargement of the mitochondria and a decrease in size of the nucleus occur in the final stages of zoospore formation. Comparison with the vegetative and sporogenous cells of other brown algae are made.

Cell Division↗

Cloning and tissue expression of 6-pyruvoyl tetrahydropterin synthase and xanthine dehydrogenase from Poecilia reticulata.

Guppy is a popular ornamental fish owing to its diverse body and fin coloration. More than 40 established color varieties have been selectively bred. The complementary DNAs for 2 enzymes that are involved in the de novo synthesis of pteridines and purines, which are important for the production of color pigments, were cloned from the caudal fin. Two cDNA isoforms for 6-pyruvoyl tetrahydropterin synthase (PTPS), with an open reading frame of 130 and 147 amino acids, respectively, were cloned from the Red Tail variety. The deduced amino acid sequence of the longer isoform shows an overall identity of about 65% to the mammalian PTPS sequences. The cDNA for xanthine dehydrogenase (XDH) was cloned from the Yellow Tail variety, and consists of an open reading frame of 1331 amino acids. Although it shows a higher overall identity to bovine aldehyde oxidase (AO; 54%) than to chicken XDH (51%), it has a NAD-binding domain that is specific to XDHs. Northern blot analysis indicated that both PTPS and XDH messenger RNAs were highly expressed in the liver, but absent in the muscle. In the caudal fins, guppy varieties with a higher proportion of xanthophores and erythrophores showed higher expression of PTPS, while XDH mRNA levels were too low to indicate obvious differential expression among the color guppy varieties. The results implied that high expression of PTPS is correlated with the biosynthesis of pteridines in the erythrophores and xanthophores, while the association between the putative guppy XDH with specific chromatophores is less clear.

Amino Acid Sequence↗

Enzymatic method for continuous monitoring of inorganic pyrophosphate synthesis.

A sensitive method for the analysis of inorganic pyrophosphate (PPi) which utilizes the enzymes ATP sulfurylase and firefly luciferase is described. The assay is based on continuous monitoring of the ATP formed in the ATP sulfurylase reaction using purified firefly luciferase. The assay can be completed in less than 2 s and is not affected by inorganic phosphate. The method has been used for continuous monitoring of formation of PPi in Rhodospirillum rubrum chromatophores. The assay is extremely sensitive, the linear range of the assay being 1 X 10(-9) - 5 X 10(-7) M PPi. It is suitable for routine applications. It is also possible to use the method for determination of low amounts of adenosine 5'-phosphosulfate.

Bacterial Chromatophores↗