Recent studies have described the role of shedding vesicles as physiological conveyers of intracellular components between neighboring cells. Here we report that melanosomes are one example of shedding vesicle cargo, but are processed by a previously unreported mechanism. Pigment globules were observed to be connected to the filopodia of melanocyte dendrites, which have previously been shown to be conduits for melanosomes. Pigment globules containing multiple melanosomes were released from various areas of the dendrites of normal human melanocytes derived from darkly pigmented skin. The globules were then captured by the microvilli of normal human keratinocytes, also derived from darkly pigmented skin, which incorporated them in a protease-activated receptor-2 (PAR-2)-dependent manner. After the pigment globules were ingested by the keratinocytes, the membrane that surrounded each melanosome cluster was gradually degraded, and the individual melanosomes then spread into the cytosol and were distributed primarily in the perinuclear area of each keratinocyte. These results suggest a melanosome transfer pathway wherein melanosomes are transferred from melanocytes to keratinocytes via the shedding vesicle system. This packaging system generates pigment globules containing multiple melanosomes in a unique manner.

Photoaging of the skin46 Solar radiation at the surface of the earth includes ultraviolet radiation (UV : 290-400nm), visible light (400-760nm) and infrared radiation (760nm-1mm) (Fig 1).Extrinsic skin aging is superimposed on intrinsic skin aging process and is due primarily to UVR (solar ultraviolet radiation) and partly by other factors, such as infrared light, smoking and air pollutants. UVR has been divided into ultraviolet B (UVB: 290-320nm) which principally generates pyrimidine dimer type DNA damage through direct absorption and ultraviolet A (UVA: 320-400nm), which indirectly produces base oxidation via UVinduced ROS.Recently , UVA radiation at high dose is reported to produce cyclobutane pyrimidine dimmers.Intrinsic aging of the skin, on the other hand, is characterized by the decline of biological function, a decrease in adaptation to stress, and structural damage due to reactive oxygen species (ROS) from cellular metabolism.Recent advances in understanding mechanisms of aging and photoaging have enhanced our ability to develop strategies to prevent, slow, and rejuvenate the altered structure and function of photoaged skin.In this review, we discuss the mechanisms of photoaging of the skin with relevance to acute and chronic skin reactions to solar UVB, UVA and infrared radiation, and summarize briefly the clinical approaches for prevention and the treatment of photoaging with topical and systemic use of anti-aging materials. Finally, a range of therapeutic modalities available to reverse or retard the visible signs of photoaged skin will be discussed briefly. There are three lights with different waveband, infrared light (IR) having waveband between 760nm and 1mm, visible light (VL) from 400nm to 760nm, and ultraviolet light (UV) from 290nm to 400nm. IR, VL and UV occupy 42 %, 52 % and 6 % of the solar light on the earth, respectively. UV light is divided into two types according to waveband, UVA having waveband between 320nm and 400nm and UVB from 290nm to 320nm. Sunburn, acute skin reaction is caused predominantly by UVB which occupies only 5~6 % of total UV light. UVA radiation, however, penetrates deeply into the dermis, around 20 % of the surface of the skin.

Proteasomes are multicatalytic proteinase complexes within cells that selectively degrade ubiquitinated proteins. We have recently demonstrated that fatty acids, major components of cell membranes, are able to regulate the proteasomal degradation of tyrosinase, a critical enzyme required for melanin biosynthesis, in contrasting manners by relative increases or decreases in the ubiquitinated tyrosinase. In the present study, we show that altering the intracellular composition of fatty acids affects the post-Golgi degradation of tyrosinase. Incubation with linoleic acid (C18:2) dramatically changed the fatty acid composition of cultured B16 melanoma cells, i.e. the remarkable increase in polyunsaturated fatty acids such as linoleic acid and arachidonic acid (C20:4) was compensated by the decrease in monounsaturated fatty acids such as oleic acid (C18:1) and palmitoleic acid (C16:1), with little effect on the proportion of saturated to unsaturated fatty acid. When the composition of intracellular fatty acids was altered, tyrosinase was rapidly processed to the Golgi apparatus from the ER (endoplasmic reticulum) and the degradation of tyrosinase was increased after its maturation in the Golgi. Retention of tyrosinase in the ER was observed when cells were treated with linoleic acid in the presence of proteasome inhibitors, explaining why melanin synthesis was decreased in cells treated with linoleic acid and a proteasome inhibitor despite the abrogation of tyrosinase degradation. These results suggest that the intracellular composition of fatty acid affects the processing and function of tyrosinase in connection with the ubiquitin-proteasome pathway and suggest that this might be a common physiological approach to regulate protein degradation.

The mechanism of melanosome transfer from melanocytes to keratinocytes has not been fully clarified. We now show a route of melanosome transfer using co-cultures of normal human melanocytes and keratinocytes. Substantial levels of melanosome transfer were elicited in co-cultures of melanocytes and keratinocytes separated by a microporous membrane filter. The melanocyte dendrites penetrated into the keratinocyte layer through the filter and many pigment globules were observed in keratinocytes. Electron microscopic observations revealed that melanosomes incorporated in keratinocytes were packed in clusters enclosed by a double membrane. Numerous pigment globules budded off from melanocyte dendrites and were released into the culture medium. Those pigment globules were filled with multiple melanosomes and a few mitochondria but no nuclei. When those globules were added to the culture medium of keratinocytes, they were incorporated and showed double membrane-enclosed melano-phagolysosomes consistent with the structures obtained from the co-culture system. In contrast, when individual naked melanosomes isolated from melanocytes were added to keratinocytes, they were also phagocytosed by keratinocytes but were enclosed by a single membrane in a manner distinct from the co-culture system. These results suggest a novel mechanism of melanosome transfer, wherein melanosomes are packed in membrane globules that bud off from melanocyte dendrites, where they are released into the extracellular space and then phagocytosed by keratinocytes.