Melanin Aggregation Research Articles

Transcriptomic Communication between Nucleus and Mitochondria during the Browning Process of Lentinula edodes.

To explore the reason for cytoplasmic replacement's significant effect on browning, transcriptomic data of nuclear (N) and mitochondrial (M) mRNAs and long noncoding RNAs (lncRNAs) in L808 and two cytoplasmic hybrids (cybrids) (L808-A2 and L808-B) of Lentinula edodes at three different culturing times (80, 100, and 120 days) were obtained. The results showed that the expression of N and M genes and lncRNAs changed with the culture time and cytoplasmic source. Cytoplasmic replacement significantly affected some M and N genes related to the internal mechanism and external morphological characteristics of L. edodes browning. The internal browning mechanism should be the nicotinamide adenine dinucleotide phosphate (NADPH)-mediated antioxidant machinery to protect mycelia against oxidative stress induced by the generation of reactive oxygen species under light irradiation. External morphological characteristics were the changing features of brown films by melanin (an antioxidant) aggregation on the surface of the mycelia of the bag or log. Especially, some genes were related to the remodeling of the plasma membrane, extracellular enzymes of celluloses and hemicellulases, small molecules, and NADPH metabolic processes. Additionally, communication between the nucleus and mitochondria mediated by M-rps3 was reported for the first time, and it is mainly appreciated in M structural assembly, functional implementation, and cooperation with other organelles.

Erbium: YAG laser treatment efficacy and association with histologic features for giant congenital melanocytic nevi management.

Limited research exists on laser treatment of giant congenital melanocytic nevus (GCMN). We sought to elucidate the efficacy of the Erbium: YAG laser on GCMN and the histologic factors associated with a positive clinical response. Between 2019 and 2022, we enrolled 30 medium-to-giant CMN patients who underwent Er: YAG laser treatment. All patients received biopsies before and after laser treatments. Clinical efficacy outcomes were evaluated by the investigator's global assessment (IGA), 5-point scale of depigmentation, and Vancouver Scar Scale (VSS) scores at least 6 months after treatment. Of the 30 cases, 18 (60.0%) showed improvement (IGA score ≥3). Eight (26.7%) patients showed repigmentation. Eight (26.7%) patients developed hypertrophic scars. The average IGA, depigmentation, and VSS scores were 2.93, 3.57, and 3.20. The IGA score was higher (3.24 ± 1.18 vs. 2.22 ± 0.97, p = 0.031) and a lower repigmentation rate (14.3% vs. 55.6%, p = 0.032) was observed in the cases with Grenz zone. The IGA score was higher (3.33 ± 1.24 vs. 2.13 ± 0.89, p = 0.023) and the repigmentation rate was lower (11.1% vs. 50.0%, p = 0.034) also in the cases with the melanocytes nests with aggregation of melanin. Lesions with superficial ablation resulted in less hypertrophic scar formation than those with deep ablation (5.9% vs. 53.8%, p < 0.05). The Er: YAG laser demonstrated effective clinical results for GCMNs. The grenz zone and the melanocytes nests with aggregation of melanin are promising predictors of laser efficacy.

Tissue regeneration effect of betulin via inhibition of ROS/MAPKs/NF-ĸB axis using zebrafish model

Betulin is the primary anti-inflammatory component of Betula platyphylla suk. cortex (birch bark), a time-honored Traditional Chinese Medicine (TCM) for healing trauma and tissue regeneration. However, the tissue regeneration effects and underlying molecular mechanism of betulin remain unknown. Therefore, it is necessary to investigate the wound repair effects and validate the mechanism of betulin in an appropriate model. In the present study, we evaluated the effects of tissue regeneration, melanin scavenging, and reactive oxygen species (ROS) inhibition of betulin using a zebrafish model. The mechanism of target genes and pathways were confirmed using quantitative polymerase chain reaction and western blotting in vivo, while molecular docking, absorption, distribution, metabolism, excretion, and toxicity investigations in-silico were conducted. Betulin significantly promoted the regeneration of zebrafish caudal fin length and area and alleviated melanin aggregation, as well as ROS generation. The relative mRNA expression of IL-1β, TNF-α, p38α, ERK1/2, and Caspase3, and the relative protein expression of p38α, ERK1/2, Caspase3, phosphorylated proteins of p-p38α, p-ERK1/2, and p-p65 were down-regulated following betulin administration. Meanwhile, the protein ratios of p-p38α/p38α, p-ERK/ERK, and p-p65/p65 were significantly decreased. In an in-silico study, binding affinities between betulin and P38α, ERK1, ERK2, and Caspase3, and the pharmacokinetic profile of betulin were predicted. The findings suggest that the tissue regeneration mechanism of betulin is based on the inhibition of excessive inflammatory responses, melanin aggregation, and the pro-apoptotic factor, Caspase3, during the proliferation phase via the ROS/MAPKs/NF-ĸB signaling axis. Our results suggest betulin as a potential candidate for tissue regeneration.

The pathogenesis of melasma and implications for treatment.

Melasma is a complex and poorly understood disorder, with high rates of treatment failure and recurrences. We aimed to review the current knowledge of the pathogenesis of melasma and apply this knowledge to clinical implications on relevant therapeutic interventions. A systematic PubMed search was performed using the search term "((melasma[Text Word]) OR facial melanosis[Text Word]) AND (pathogenesis OR causality[MeSH Terms])" for articles published between 1990 and 2020. Included articles were then evaluated by two authors and assessed for relevant pathomechanistic pathways, after which they were divided into groups with minimal overlap. We then reviewed current treatment modalities for melasma and divided them according to the involved pathomechanistic pathway. A total of 309search results were retrieved among which 76 relevant articles were identified and reviewed. Five main pathomechanisms observed in melasma were identified: (1) melanocyte inappropriate activation; (2) aggregation of melanin and melanosomes in dermis and epidermis; (3a) increased mast cell count and (3b) solar elastosis; (4) altered basement membrane; and (5) increased vascularization. Treatment modalities were then divided based on these five pathways and detailed in 6 relevant tables. The pathophysiology of melasma is multifactorial, resulting in treatment resistance and high recurrence rates. This wide variety of pathomechanisms should ideally be addressed separately in the treatment regimen in order to maximize results.

The effect of static magnetic field on melanophores in the sea trout (Salmo trutta m. trutta Linnaeus, 1758) embryos and larvae

AbstarctMagnetic field affects many living organisms, fishes included. The fish reactions are observed from the early stage of development, during embryogenesis and larval period, and into adult stage. Magnetic field affects, among other processes, water exchange in the eggs, embryo heart rate and respiration, directional reactions and spatial orientation. The few papers in this field published so far present melanophores morphology and development. Melanophores developing in embryos and in newly hatched sea trout (Salmo trutta m. trutta L.) larvae which had been incubated and kept under optimal conditions constituted the material for the study. The effect of static, generated magnetic field of intensity of 1, 3 and 5 mT on melanophores of sea trout embryos and larvae was studied. The control setting was kept under geomagnetic field. The appearance of melanophores in the trout embryos exposed to magnetic field was delayed. The number of melanophores was smaller, and the movement of melanin within the melanophores, visible as aggregation of pigment in the cells. Static magnetic field of relatively low intensity influences the reaction of melanophores of developing trout embryos and newly hatched larvae. Most probably, magnetite (Fe3O4) compounds, detected in many fish species, mainly migratory fishes such as trout, are responsible for such behaviour of melanophores.HighlightsStatic magnetic field delays the appearance of melanophores in the trout embryos and larvae.The largest aggregation of melanin in melanophores was found in larvae incubated in a 5 mT magnetic field.The largest number of melanophores was found in the embryos incubated in the geomagnetic field.

ASIP disruption via CRISPR/Cas9 system induces black patches dispersion in Oujiang color common carp

Pigment pattern formation in economical fish remains elusive to date. Oujiang color common carp (Cyprinus carpio var. color) is a variant of common carp with different pigmentation patterns and important farm fish in China. Agouti signaling protein (ASIP) is an endogenous antagonist of melanocortin in the melanogenesis pathway. In this study, two ASIP genes (ASIP 1 and ASIP 2) were first identified in color common carp. Their expression was higher in the pale belly than in the dark dorsal skin, which was consistent with the typical dorsal–ventral expression pattern such as that of wild common carp. In comparison, an unusually higher expression was detected in the non-melanophore side skin than in the melanophore side skin of Oujiang color common carp. Thus, CRISPR/Cas9 was used to disrupt the two ASIP genes in Oujiang color common carp of red with big black patches (RB) pattern. Consequently, the black patches disappeared and the melanophores dispersed along the dorsal skin in the F0 mosaic RB individuals. The findings of this study indicate that the ASIP gene is involved in regulating melanin aggregation and distribution during black patch formation in Oujiang color common carp.

Effect of cadmium chloride on general body colouration and chromatophores of stinging cat fish, Heteropneustes fossilis (Bloch)

Chromatophores, specialized pigment cells in poikilothermic animals, have shown great potential in their use as a cell-based biosensor in the detection of a broad range of environmental toxicants, as structure and number of chromatophores alters significantly under toxicant exposure. Skin coloration of Heteropneustes fossilis is due to melanin containing melanophores. Cadmium, the black listed and non essential heavy metal, is widely used that adversely affects vital activities of aquatic biota. H. fossilis, freshwater Indian stinging catfish, were subjected to exposure of 96 hour LC50 dose (392.92 mg/l) and 25% of 96 hour LC50 dose (98.23mg/l) of cadmium chloride (CdCl2) to evaluate toxic impact of cadmium on colouration and chromatophores. A significant decrease was observed in number of chromatophores after acute (highly significant (F = 70.50; P&lt;0.001) and sub acute (significant (F = 0.29; P&lt;0.05) exposure along with heavy nacrotic, lytic and degenerative changes. Chromatophore gradually changed from reticulate to punctate-stellate and punctuate type as they lost their dendritic processes and aggregation of melanin towards centre. Most of the chromatophores lost their cellular entity due to degenerative changes and melanin was found dispersed in surrounding matrix. Peeling and fading of skin was the common feature in all exposure durations. Fish chromatophores may serve as better biomarkers in reference to metallic pollution and will also be helpful in accessing the health status of economically important fishes as well as worsening status of aquatic bodies.

Genomewide analysis of copy number variants in alopecia areata in a Central European cohort reveals association with MCHR2.

Alopecia areata (AA) is a common hair loss disorder of autoimmune aetiology, which often results in pronounced psychological distress. Understanding of the pathophysiology of AA is increasing, due in part to recent genetic findings implicating common variants at several genetic loci. To date, no study has investigated the contribution of copy number variants (CNVs) to AA, a prominent class of genomic variants involved in other autoimmune disorders. Here, we report a genomewide- and a candidate gene-focused CNV analysis performed in a cohort of 585 patients with AA and 1340 controls of Central European origin. A nominally significant association with AA was found for CNVs in the following five chromosomal regions: 4q35.2, 6q16.3, 9p23, 16p12.1 and 20p12.1. The most promising finding was a 342.5-kb associated region in 6q16.3 (duplications in 4/585 patients; 0/1340 controls). The duplications spanned the genes MCHR2 and MCHR2-AS1, implicated in melanin-concentrating hormone (MCH) signalling. These genes have not been implicated in previous studies of AA pathogenesis. However, previous research has shown that MCHR2 affects the scale colour of barfin flounder fish via the induction of melanin aggregation. AA preferentially affects pigmented hairs, and the hair of patients with AA frequently shows a change in colour when it regrows following an acute episode of AA. This might indicate a relationship between AA, pigmentation and MCH signalling. In conclusion, the present results provide suggestive evidence for the involvement of duplications in MCHR2 in AA pathogenesis.

Environmental concentrations of prednisolone alter visually mediated responses during early life stages of zebrafish (Danio rerio)

The development of the eye in vertebrates is dependent upon glucocorticoid signalling, however, specific components of the eye are sensitive to synthetic glucocorticoids. The presence of synthetic glucocorticoids within the aquatic environment may therefore have important consequences for fish, which are heavily reliant upon vision for mediating several key behaviours. The potential ethological impact of synthetic glucocorticoid oculotoxicity however has yet to be studied. Physiological and behavioural responses which are dependent upon vision were selected to investigate the possible toxicity of prednisolone, a commonly occurring synthetic glucocorticoid within the environment, during early life stages of zebrafish. Although exposure to prednisolone did not alter the morphology of the external eye, aggregation of melanin within the skin in response to increasing light levels was impeded and embryos exposed to prednisolone (10 μg/l) maintained a darkened phenotype. Exposure to prednisolone also increased the preference of embryos for a dark environment within a light dark box test in a concentration dependent manner. However the ability of embryos to detect motion appeared unaffected by prednisolone. Therefore, while significant effects were detected in several processes mediated by vision, changes occurred in a manner which suggest that vision was in itself unaffected by prednisolone. Neurological and endocrinological changes during early ontogeny are considered as likely candidates for future investigation.

Melanopsin photoreception in the eye regulates light-induced skin colour changes through the production of α-MSH in the pituitary gland.

How skin colour adjusts to circadian light/dark cycles is poorly understood. Melanopsin (Opn4) is expressed in melanophores, where in vitro studies suggest it regulates skin pigmentation through a 'primary colour response' in which light photosensitivity is translated directly into pigment movement. However, the entrainment of the circadian rhythm is regulated by a population of melanopsin-expressing retinal ganglion cells (mRGCs) in the eye. Therefore, in vivo, melanopsin may trigger a 'secondary colour response' initiated in the eye and controlled by the neuro-endocrine system. We analysed the expression of opn4m and opn4x and melanin aggregation induced by light (background adaptation) in Xenopus laevis embryos. While opn4m and opn4x are expressed at early developmental times, light-induced pigment aggregation requires the eye to become functional. Pharmacological inhibition of melanopsin suggests a model whereby mRGC activation lightens skin pigmentation via a secondary response involving negative regulation of alpha-melanocyte-stimulating hormone (α-MSH) secretion by the pituitary.

Near-Infrared Excited State Dynamics of Melanins: The Effects of Iron Content, Photo-Damage, Chemical Oxidation, and Aggregate Size

Ultrafast pump–probe measurements can discriminate the two forms of melanin found in biological tissue (eumelanin and pheomelanin), which may be useful for diagnosing and grading melanoma. However, recent work has shown that bound iron content changes eumelanin’s pump–probe response, making it more similar to that of pheomelanin. Here we record the pump–probe response of these melanins at a wider range of wavelengths than previous work and show that with shorter pump wavelengths the response crosses over from being dominated by ground-state bleaching to being dominated by excited-state absorption. The crossover wavelength is different for each type of melanin. In our analysis, we found that the mechanism by which iron modifies eumelanin’s pump–probe response cannot be attributed to Raman resonances or differences in melanin aggregation and is more likely caused by iron acting to broaden the unit spectra of individual chromophores in the heterogeneous melanin aggregate. We analyze the dependence on optical intensity, finding that iron-loaded eumelanin undergoes irreversible changes to the pump–probe response after intense laser exposure. Simultaneously acquired fluorescence data suggest that the previously reported “activation” of eumelanin fluorescence may be caused in part by the dissociation of metal ions or the selective degradation of iron-containing melanin.

Identification of mRNAs coding for mammalian-type melanin-concentrating hormone and its receptors in the scalloped hammerhead shark Sphyrna lewini

Melanin-concentrating hormone (MCH) is a neuromodulator, synthesized in the hypothalamus, that regulates both appetite and energy homeostasis in mammals. MCH was initially identified in teleost fishes as a pituitary gland hormone that induced melanin aggregation in chromatophores in the skin; however, this function of MCH has not been observed in other vertebrates. Recent studies suggest that MCH is involved in teleost feeding behavior, spurring the hypothesis that the original function of MCH in early vertebrates was appetite regulation. The present study reports the results of cDNAs cloning encoding preproMCH and two MCH receptors from an elasmobranch fish, Sphyrna lewini, a member of Chondrichthyes, the earliest diverged class in gnathostomes. The putative MCH peptide is composed of 19 amino acids, similar in length to the mammalian MCH. Reverse-transcription polymerase chain reaction revealed that MCH is expressed in the hypothalamus in S. lewini MCH cell bodies and fibers were identified by immunochemistry in the hypothalamus, but not in the pituitary gland, suggesting that MCH is not released via the pituitary gland into general circulation. MCH receptor genes mch-r1 and mch-r2 were expressed in the S. lewini hypothalamus, but were not found in the skin. These results indicate that MCH does not have a peripheral function, such as a melanin-concentrating effect, in the skin of S. lewini hypothalamic MCH mRNA levels were not affected by fasting, suggesting that feeding conditions might not affect the expression of MCH in the hypothalamus.

Effect of gamma irradiation on biophysical and protection properties of melanin

Melanins are natural pigments distributed in living organisms and they are responsible for pigmentation of surface structure. In the present work, gel exclusion chromatography, spectrophotometric, and dielectric relaxation techniques were used to characterize DOPA melanin before and after irradiation with 60Co gamma rays in the range of 5 to 50 Gy. The results show that the studied melanin is composed of two structural groups. The extinction absorption coefficient and relative permittivity and conductivity of melanin sample increased after irradiation with gamma rays and were dose dependent in manner. These increases were attributed to the formation of melanin aggregation and crosslinks which result from the growing number of the formed free radicals by radiation. It was concluded from the results that melanin goes through some structural changes after irradiation with the gamma doses demonstrated. Further studies were recommended to investigate and evaluate whether these changes could affect its efficiency as a radio protector against gamma radiation. Key words: Melanin structure, gamma irradiation, gel chromatography absorption spectra, relative permittivity and conductivity.

On The Novel Action of Melanolysis by a Leaf Extract of Aloe vera and Its Active Ingredient Aloin, Potent Skin Depigmenting Agents

The present study was carried out to investigate the effects of an Aloe vera leaf extract, along with its standard active ingredient aloin, on the isolated tail melanophores of Bufo melanostictus tadpoles, which are a type of disguised smooth muscle cells offering excellent in vitro opportunities for studying the effects of pharmacological and pharmaceutical agents. It was found that the leaf extract of A.vera and its active ingredient aloin induced powerful, dose-dependent, physiologically significant melanin aggregating effects in the isolated tail melanophores of B. melanostictus similar to those of adrenaline per se. These preliminary findings clearly demonstrate that the extract of A.vera and its active ingredient aloin cause melanin aggregation leading to skin lightening via alpha adrenergic receptor stimulation. The present study opens new vistas for the use of A.vera regarding its clinical application as a new nontoxic melanolytic agent for the treatment of hyperpigmentation.

Molecular cloning and expression of two melanin-concentrating hormone receptors in goldfish

Melanin-concentrating hormone (MCH) is a neurohypophysial hormone and induces melanin aggregation in the skin in teleosts. MCH also has multiple roles in the central regulation of food intake in teleosts and mammals. MCH receptors (MCH-R) are among type I G-protein-coupled receptors. Here, we cloned two MCH receptors from goldfish, Carassius auratus. The amino acid sequence of goldfish MCH-R1 had 57–88% homology with fish MCH-R1 and 49–50% homology with mammalian MCH-R1, while the amino acid sequence of goldfish MCH-R2 had 72–92% homology with fish MCH-R2 and 32% homology with human MCH-R2. Phylogenetic analysis showed that these two MCH-Rs are orthologous to the respective mammalian MCH-Rs. The common amino acid residues for ligand binding, signal transduction, and receptor conformation were well conserved in these receptors, although some intracellular basic-amino-acid-rich domains, which have been shown to exist in human MCH-R1 and MCH-R2, were absent in goldfish MCH-R2. When stably expressed in HEK293 cells, both goldfish MCH-R1 and MCH-R2 displayed a strong, dose-dependent, transient elevation of intracellular calcium in response to salmon MCH (EC 50 = 0.8 nM and 31.8 nM, respectively). In contrast to goldfish MCH-R2, goldfish MCH-R1 signaling is not sensitive to pertussis toxin, suggesting an exclusive Gαq coupling of goldfish MCH-R1 in the mammalian cell-based assay. Reverse transcriptase PCR revealed that both MCH-R1 and MCH-R2 mRNA are distributed in various tissues in goldfish. The various tissues including the brain and skin express both MCH-R1 and MCH-R2. These results suggest that these functional receptors mediate multiple effects of MCH in goldfish.

Mc1rPathway Regulation of Zebrafish Melanosome Dispersion

Zebrafish rapidly alter their pigmentation in response to environmental changes. For black melanocytes, this change is due to aggregation or dispersion of melanin in the cell. Dispersion and aggregation are controlled by intracellular cyclic adenosine monophosphate (cAMP) levels, which increase upon stimulation by alpha melanocyte-stimulating hormone (alpha-MSH) or reduce with melanin-concentrating hormone (MCH). In mammals and birds, the melanocortin-1-receptor (MC1R) responds to MSH, and stimulates the synthesis of black eumelanin. While MSH-cAMP signaling stimulates melanogenesis in mammals, and melanosome dispersal in cold-blood vertebrates, the pathway components are highly conserved. However, it has only been assumed that mc1r mediates melanosome dispersal in fish. Here, using morpholino oligonucleotides designed to knockdown mc1r expression, we find that mc1r morphants are unable to disperse melanosomes when grown in dark conditions. We also use chemical modifiers of the cAMP pathway, and find an unexpected response to the specific phosphodiesterase 4 (PDE4) inhibitor, rolipram, in melanosome dispersal. When treated with the drug, melanosomes fail to fully disperse in dark conditions, despite presumed increased levels of cAMP, and in contrast to the effects of the nonselective PDE inhibitor, 3-isobutyl-1-methylxanthine. In conclusion, we demonstrate a direct role for mc1r in zebrafish melanosome dispersal in response to background, and use chemical modification of this pathway to uncover a possible new layer of regulation in melanosome dispersal in zebrafish.

Melanin photoprotection in the human retinal pigment epithelium and its correlation with light-induced cell apoptosis

Time-resolved electron paramagnetic resonance (TREPR) spectroscopy was used to study melanin free radicals in human retinal pigment epithelium (RPE) cells and tyrosine-derived synthetic melanin. TREPR signal traces from RPE cells reveal in vivo light-induced melanin free radical photochemistry in more detail than previously known. Electron spin polarization reflecting a non-Boltzmann population within the energy levels of the spin system is observed in RPE cells as the result of the triplet state photoproduction and subsequent disappearance of free radicals in the melanin polymer. In a set of RPE cells cultured from individual sources, differences in optical absorption, continuous wave EPR spectra, and TREPR signals were correlated with apoptosis assays performed by flow cytometry. Continuous wave EPR spectra of RPE cells and TREPR of acidified synthetic melanin suggest that increased melanin aggregation provides an increase in photoprotection in the RPE cells that are relatively less susceptible to blue light-induced apoptosis.

Isolation and biophysical studies of natural eumelanins: applications of imaging technologies and ultrafast spectroscopy.

The major pigments found in the skin, hair, and eyes of humans and other animals are melanins. Despite significant research efforts, the current understanding of the molecular structure of melanins, the assembly of the pigment within its organelle, and the structural consequences of the association of melanins with protein and metal cations is limited. Likewise, a detailed understanding of the photochemical and photophysical properties of melanins has remained elusive. Many types of melanins have been studied to date, including natural and synthetic model pigments. Such studies are often contradictory and to some extent the diversity of systems studied may have detracted from the development of a basic understanding of the structure and function of the natural pigment. Advances in the understanding of the structure and function of melanins require careful characterization of the pigments examined so as to assure the data obtained may be relevant to the properties of the pigment in vivo. To address this issue, herein the influence of isolation procedures on the resulting structure of the pigment is examined. Sections describing the applications of new technologies to the study of melanins follow this. Advanced imaging technologies such as scanning probe microscopies are providing new insights into the morphology of the pigment assembly. Recent photochemical studies on photoreduction of cytochrome c by different mass fraction of sonicated natural melanins reveal that the photogeneration of reactive oxygen species (ROS) depends upon aggregation of melanin. Specifically, aggregation mitigates ROS photoproduction by UV-excitation, suggesting the integrity of melanosomes in tissue may play an important role in the balance between the photoprotective and photodamaging behaviors attributed to melanins. Ultrafast laser spectroscopy studies of melanins are providing insights into the time scales and mechanisms by which melanin dissipates absorbed light energy.

The Photoreceptor Molecules in Xenopus Tadpole Tail Fin, in which Melanophores Exist

Abstract Melanophores of the isolated tail fin of the Xenopus tadpole respond to light, resulting in melanin aggregation in the melanophores. Western blot analysis showed that a protein in the Xenopus tail fins, in which photosensitive melanophores exist, had reacted with the antibody against bovine rhodopsin. RT-PCR and nested-PCR using rhodopsin-specific primers showed the expression of rhodopsin mRNA in the tail fins. The amino acid sequences deduced from the PCR products were completely identical to those of rhodopsin. We also detected the mRNA of melanopsin in the tail fin, another opsin originally described in cultured melanophores of Xenopus. These results indicate that these two types of opsin molecules exist in Xenopus tail fin and may take part in the photo-response in melanophores of the Xenopus tadpole.

Solution Structure of Copper Ion-Induced Molecular Aggregates of Tyrosine Melanin

Melanin, the ubiquitous biological pigment, provides photoprotection by efficient filtration of light and also by its antioxidant behavior. In solutions of synthetic melanin, both optical and antioxidant behavior are affected by the aggregation states of melanin. We have utilized small-angle x-ray and neutron scattering to determine the molecular dimensions of synthetic tyrosine melanin in its unaggregated state in D 2O and H 2O to study the structure of melanin aggregates formed in the presence of copper ions at various copper-to-melanin molar ratios. In the absence of copper ions, or at low copper ion concentrations, tyrosine melanin is present in solution as a sheet-like particle with a mean thickness of 12.5 Å and a lateral extent of ∼54 Å. At a copper-to-melanin molar ratio of 0.6, melanin aggregates to form long, rod-like structures with a radius of 32 Å. At a higher copper ion concentration, with a copper-to-melanin ratio of 1.0, these rod-like structures further aggregate, forming sheet-like structures with a mean thickness of 51 Å. A change in the charge of the ionizable groups induced by the addition of copper ions is proposed to account for part of the aggregation. The data also support a model for the copper-induced aggregation of melanin driven by π stacking assisted by peripheral Cu 2+ complexation. The relationship between our results and a previous hypothesis for reduced cellular damage from bound-to-melanin redox metal ions is also discussed.