The epidermal melanin unit in human skin is composed of melanocytes and keratinocytes. Melanocytes, located in the basal layer of the epidermis, manufacture melanin-loaded organelles called melanosomes. Through their dendritic processes, melanocytes distribute melanosomes to neighboring keratinocytes, where their presence confers to the skin its characteristic color and photoprotective properties. In this study, we used murine melanocytes and keratinocytes alone and in coculture to characterize the processes involved in melanosome transfer. Ultraviolet (UV) radiation induced an accumulation of melanosomes in melanocytes, whereas treatment with a-melanocyte-stimulating hormone (MSH) induced exocytosis of melanosomes accompanied by ruffling of the melanocyte membrane. We found that keratinocytes phagocytose melanosomes and latex beads equally well and that this phagocytic process was increased by exposure of keratinocytes to UV radiation or to MSH. Coculture of melanocytes and keratinocytes resulted in an increase in MSH released to the medium. Gene array analysis of MSH-treated melanocytes showed up-regulation of many genes associated with exocytosis. In our studies, we never observed cytophagocytosis of melanosome-filled processes. This result, together with the other findings, suggests that a combination of signals that increase melanosome production and release by melanocytes and that stimulate phagocytosis by keratinocytes are the most relevant mechanisms involved in skin tanning.
The two main pigment types in bird feathers are the red, orange, and yellow carotenoids and the black, gray, and brown melanins. Reports conflict, however, regarding the potential for melanins to produce yellow colors or for carotenoids to produce brown plumages. We used high-performance liquid chromatography to analyze carotenoids and melanins present in the yellow and brown feathers of five avian species: Eastern Bluebirds (Sialia sialis), Barn Swallows (Hirundo rustica), King Penguins (Aptenodytes patagonicus), Macaroni Penguins (Eudyptes chrysolophus), and neonatal chickens (Gallus domesticus). In none of these species did we detect carotenoid pigments in feathers. Although carotenoids are reportedly contained in the ventral plumage of European Barn Swallows (Hirundo rustica rustica), we instead found high concentrations of both eumelanins and phaeomelanins in North American Barn Swallows (H. r. erythrogaster). We believe we have detected a new form of plumage pigment that gives penguin and domestic- chick feathers their yellow appearance. No Puedes Juzgar un Pigmento por su Color: Contenido de Carotenoide y Melanina de Plumas Amarillas y Marrones en Golondrinas, Azulejos, Pingüinos y Gallinas Domésticas Resumen. Los dos tipos principales de pigmentos que las aves incorporan en sus plumas son carotenoides, para desarrollar plumajes rojo, naranja o amarillo, y melaninas, para adquirir coloración negra, marrón, gris o tonalidades color tierra. Sin embargo, existe información conflictiva sobre la potencial coloración de plumas amarillas basadas en melanina y la presencia de caroteniodes en el plumaje marrón de ciertas especies. En este estudio, usamos cromatografía líquida de alto rendimiento para analizar los tipos y cantidades de carotenoides y melaninas presentes en las plumas amarillas y marrones de cinco especies de aves: el azulejo Sialia sialis y la golondrina Hirundo rustica, los pingüinos Aptenodytes patagonicus y Eudyptes chrysolophus y el plumón natal amarillo de la gallina doméstica Gallus domesticus. En ninguna de estas especies detectamos pigmentos carotenoides en las plumas. A pesar de que los carotenoides han sido encontrados en el plumaje ventral de la golondrina Hirundo rustica rustica, nosotros en cambio encontramos altas concentraciones de eumelaninas y feomelaninas en H. r. erythrogaster y en azulejos que variaron entre individuos y regiones de plumaje. Creemos que hemos detectado una nueva forma de pigmento de plumaje que le da a las plumas de pingüinos y pollos domésticos su apariencia amarilla.
Melanosomes were isolated from the retinal pigment epithelium (RPE), iris and choroid of mature (age >2 years) and newborn (age <1 week) bovine eyes. Scanning electron microscopy was utilized to analyze the morphology of the melanosomes, which were found to vary among different tissues and different ages. While the total content of amino acids differs slightly (ranging from 9% to 15% by mass), the distributions of the amino acids are similar. The pheomelanin content is low in the choroid and the RPE (0.1–0.5%), and moderate in the iris (<2%); therefore, the major melanin component of bovine eye melanosomes is eumelanin, independent of the shape of the melanosomes. The yields of pyrrole‐2,3,5‐tricarboxylic acid from melanosomes decrease in the following order: choroid > iris > RPE, and exhibit decreasing yields with age. 13C solid‐state nuclear magnetic resonance (NMR) spectroscopic analysis of iris and choroid melanosomes indicates the same trends. These observations suggest that the 5,6‐dihydroxyindole‐2‐carboxylic acid contents decrease in the following order: choroid > iris > RPE, and decrease with age. Moreover, the 13C solid‐state NMR spectra show (1) for the same age samples, the CH:Cq ratio for choroid is larger than that for iris melanosomes; and (2) an increase in the concentration of carbonyl groups with age within each type of melanosome.
Cutaneous pigmentation is determined by the amounts of eumelanin and pheomelanin synthesized by epidermal melanocytes and is known to protect against sun-induced DNA damage. The synthesis of eumelanin is stimulated by the binding of α-melanotropin (α-melanocyte-stimulating hormone)to the functional melanocortin 1 receptor (MC1R) expressed on melanocytes. The human MC1R gene is highly polymorphic and certain allelic variants of the gene are associated with red hair phenotype, melanoma and non-melanoma skin cancer. The importance of the MC1R gene in determining skin cancer risk led us to examine the impact of specific polymorphisms in this gene on the responses of human melanocytes to α-melanotropin and UV radiation. We compared the ability of human melanocyte cultures, each derived from a single donor, to respond to α-melanotropin with dose-dependent stimulation of cAMP formation, tyrosinase activity and proliferation. In each of those cultures the MC1R gene was sequenced, and the eumelanin and pheomelanin contents were determined. Human melanocytes homozygous for Arg160Trp, heterozygous for Arg160Trp and Asp294His, or for Arg151Cys and Asp294His substitutions, but not melanocytes homozygous for Val92Met substitution, in the MC1R demonstrated a significantly reduced response toα-melanotropin. Additionally, melanocytes with a non-functional MC1R demonstrated a pronounced increase in their sensitivity to the cytotoxic effect of UV radiation compared with melanocytes expressing functional MC1R. We conclude that loss-of-function mutations in the MC1R gene sensitize human melanocytes to the DNA damaging effects of UV radiation, which may increase skin cancer risk.
In order to develop melanoma-targeted <i>in situ</i> peptide vaccine immunotherapy, magnetite nanoparticles were conjugated with a melanogenesis substrate, N-propionyl cysteaminylphenol (NPrCAP). Magnetite nanoparticles introduced thermotherapy which caused non-apoptotic cell death and generation of heat shock protein (HSP) upon exposure to alternating magnetic field (AMF). NPrCAP was expected to develop a melanoma-targeted therapeutic drug because of its selective incorporation into melanoma cells and production of highly reactive free radicals, that result in not only oxidative stress but also apoptotic cell death by reacting with tyrosinase
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