Hair Follicular Melanocytes Research Articles

Worldwide cutaneous malignant melanoma incidences analyzed by sex, age, and skin type over time (1955-2007): Is HPV infection of androgenic hair follicular melanocytes a risk factor for developing melanoma exclusively in people of European-ancestry?

ABSTRACTThe cutaneous malignant melanoma (CMM) incidence has been increasing in an exponential manner in certain populations around the world for over 7 decades. To help illuminate the etiology, we performed worldwide temporal (1955–2007) CMM incidence analysis by sex, age (0–14, 15–29, 30–49, 50–69, 70–85+), and skin type on 6 continents using data from the International Agency for Research on Cancer. We observe an exponential increase in the CMM incidence over time and an increase of about 2 orders of magnitude between age groups 0–14 and 15–29 exclusively in European-ancestry populations around the world independent of skin type (I–III or III–IV). Other populations like the Chinese (III-IV) had much lower CMM incidences that either remained stable or temporally decreased but did not display a dramatic increase between the youngest age groups. The dramatic increase in the incidence between the youngest age groups found only in European-ancestry populations suggests one of the most important risk factors for CMM may be developing androgenic hair, the occurrence of which appears to correlate with the distribution of CMM over male and female body sites. Besides that potential new risk factor, the increasing CMM incidence with increasing age, known not to be from cumulative UV doses, may be associated with age-related changes to skin, i.e., thinning epidermis causing lower vitamin D3 levels, and hair, i.e., whitening from higher reactive oxygen species. The temporal exponential increasing CMM incidence in European-ancestry populations may be due to Human Papilloma Virus infection of follicular hair melanocytes, found in CMM biopsies.

The biology of hair diversity

Hair diversity, its style, colour, shape and growth pattern is one of our most defining characteristics. The natural versus temporary style is influenced by what happens to our hair during our lifetime, such as genetic hair loss, sudden hair shedding, greying and pathological hair loss in the various forms of alopecia because of genetics, illness or medication. Despite the size and global value of the hair care market, our knowledge of what controls the innate and within-lifetime characteristics of hair diversity remains poorly understood. In the last decade, drivers of knowledge have moved into the arena of genetics where hair traits are obvious and measurable and genetic polymorphisms are being found that raise valuable questions about the biology of hair growth. The recent discovery that the gene for trichohyalin contributes to hair shape comes as no surprise to the hair biologists who have believed for 100years that hair shape is linked to the structure and function of the inner root sheath. Further conundrums awaiting elucidation include the polymorphisms in the androgen receptor (AR) described in male pattern alopecia whose location on the X chromosome places this genetic contributor into the female line. The genetics of female hair loss is less clear with polymorphisms in the AR not associated with female pattern hair loss. Lifestyle choices are also implicated in hair diversity. Greying, which also has a strong genetic component, is often suggested to have a lifestyle (stress) influence and hair follicle melanocytes show declining antioxidant protection with age and lowered resistance to stress. It is likely that hair research will undergo a renaissance on the back of the rising information from genetic studies as well as the latest contributions from the field of epigenetics.

Functionally distinct melanocyte populations revealed by reconstitution of hair follicles in mice

Hair follicle reconstitution analysis was used to test the contribution of melanocytes or their precursors to regenerated hair follicles. In this study, we first confirmed the process of chimeric hair follicle regeneration by both hair keratinocytes and follicular melanocytes. Then, as first suggested from the differential growth requirements of epidermal skin melanocytes and non-cutaneous or dermal melanocytes, we confirmed the inability of the latter to be involved as follicular melanocytes to regenerate hair follicles during the hair reconstitution assay. This clear functional discrimination between non-cutaneous or dermal melanocytes and epidermal melanocytes suggests the presence of two different melanocyte cell lineages, a finding that might be important in the pathogenesis of melanocyte-related diseases and melanomas.

Estrogen Increases Hair Pigmentation in Female Recessive Yellow Mice

Murine recessive yellow (Mc1r(e)) is a loss-of-function mutation in the receptor for alpha-melanocyte-stimulating hormone, melanocortin receptor 1 (Mc1r), and results in a yellow coat by inducing pheomelanin synthesis in hair follicular melanocytes. We previously showed that eumelanin and pheomelanin content in dorsal hair in female Mc1r(e)/Mc1r(e) mice 5 weeks after birth was greater than that in male mice. To better understand the sex difference in hair pigmentation, estrogen, progesterone, and androgen were injected subcutaneously to newborn mice, and the effects of these sex hormones on hair pigmentation at 8.5 days were investigated. Although these sex hormones failed to affect the ratio of pheomelanic to eumelanic hair, they increased total hair pigmentation. Chemical analyses showed that total melanin content was increased by a low dose of estrogen in female mice. Moreover, estrogen increased pheomelanin content in female hair, whereas the hormone greatly decreased both eumelanin and pheomelanin content in male hair. High doses of progesterone, in contrast, increased total melanin content in both female and male hair. Moreover, a high dose of androgen increased total melanin content exclusively in male hair. These results suggest that estrogen is the main factor in determining the higher content of eumelanin and pheomelanin in female hair of Mc1r(e)/Mc1r(e) mice.

Excess tyrosine rescues the reduced activity of proliferation and differentiation of cultured recessive yellow melanocytes derived from neonatal mouse epidermis

The murine recessive yellow ( Mc1r e ) is a loss-of-function mutation in the receptor for α-melanocyte-stimulating hormone, melanocortin receptor 1 (Mc1r) and produces yellow coats by inducing pheomelanin synthesis in hair follicular melanocytes. However, it is not known whether the Mc1r e mutation affects the proliferation and differentiation of melanocytes. In this study, the proliferation and differentiation of recessive yellow epidermal melanocytes cultured in dibutyryl cyclic AMP-supplemented serum-free medium were investigated in detail. The melanocytes produced mainly eumelanin in this culture system. The proliferation of recessive yellow melanocytes was decreased compared with that of wild-type at the e-locus, black melanocytes. The differentiation of melanocytes was also delayed and inhibited in recessive yellow mice. Tyrosinase (TYR) activity and TYR-related protein 1 (TRP1) and TRP2 (dopachrome tautomerase, DCT) expressions were decreased and, in addition, the maturation of stage IV melanosomes was inhibited. Excess l-tyrosine ( l-Tyr) added to the culture media rescued the reduced activity of proliferation of melanocytes. l-Tyr also stimulated TYR activity and TRP1 and TRP2 expressions as well as the maturation of stage IV melanosomes and pigmentation. These results suggest that the Mc1r e mutation affects the proliferation and differentiation of melanocytes and l-Tyr rescues the reduced proliferative and differentiative activities by stimulating TYR activity and TRP1 and TRP2 expressions as well as melanosome maturation.

The Hair Follicle: A Specialised UV Receptor in the Human Skin?

Coat colour changes in polar animals are related to seasonal variation in photic inputs. The present work was performed to study the photoresponses of hair follicular melanocytes in human skin. The melanocytes, being photosensitive cells, can function as UV biosensors, since dendrites extend towards the source of UV light. Fifty-one skin biopsies from the margin of vitiligo were subjected to whole skin organ cultures. These were exposed to a pulse of UV light to study hair bulb melanocytes in vitiligo. It is observed that the melanocytes are seen within the anagen matrix. These melanocytes are poorly dendritic in control and dark-incubated cultures. On UV exposure, they become highly dendritic, the dendrites extending towards the hair shaft in 93.5%. They show prominent catechol oxidase and noradrenaline positivity, all features of UV responsiveness. The melanocytes within the hair follicle are not directly exposed to UV light. The melanocyte dendricity and the alignment of dendrites towards the shaft on UV exposure indicate that the columns of the cells in the hair shaft act as an efficient fibre-optic system, transmitting UV light. Morphologically, the keratinocytes in the hair shaft are arranged in compressed linear columns which resemble the coaxial bundles of commercial fibre-optic strands as is observed in plants. Keratinocytes in the inner and outer sheaths do not show this arrangement. Thus the hair follicle functions as a specialised UV receptor in the skin responding to nuances of photic inputs in human skin. This is reflected in coat colour changes in animals exposed to large variations in day-night cycles.

Proliferating Activity of the Hair Follicular Melanocytes at the Early and Anagen III Stages in the Hair Growth Cycle: Detection by Immunocytochemistry for Bromodeoxyuridine Combined with DOPA Reaction Cytochemistry

In order to demonstrate the existence of proliferating activity in hair follicular melanocytes during the early and anagen III stages, immunocytochemistry for bromodeoxyuridine (BrdU) incorporated in the nuclei of the melanocytes for 1 hr was undertaken at the light and electron microscopic levels in combination with DOPA reaction cytochemistry. Light microscopically, about 30% of the nuclei of the DOPA reaction positive melanocytes at the anagen III stage of the hair growth cycle were labeled by anti-BrdU antibody. These melanocytes were observed in about one hundred hair follicles. Electron microscopically, the nuclei of a few undifferentiated melanocytes at the anagen II stage were labeled by anti-BrdU antibody-affiliated secondary antibody-gold particles. At the anagen III stage, localization of the immunogold particles in the DNA replication domains of the nuclei of the undifferentiated and redifferentiating melanocytes covered the patterns of the early, middle, and late S-phase. These findings suggest that the hair follicular melanocytes begin proliferation at the anagen II stage and that the melanocytes proliferate actively at the anagen III stage to repopulate the hair matrix during subsequent hair growth.

Effects of melanocyte-stimulating hormone on tyrosinase expression and melanin synthesis in hair follicular melanocytes of the mouse.

Tyrosinase mRNA, synthesis and activity were measured in the skin during the first 2 weeks of life of C3H-HeAvy mice. Tyrosinase mRNA levels were found to peak on days 3-4 and were followed by increases in tyrosinase synthesis and activity which peaked on days 6-7 and 7-8 respectively. These changes in tyrosinase expression were presumably associated with the growth of the first coat of hair that in neonatal C3H-HeAvy mice is yellow in colour as a result of the increased proportion of phaeomelanin. By the time hair growth had ceased there was no expression of tyrosinase at both mRNA and protein levels. Daily administration of alpha-melanocyte stimulating hormone (alpha-MSH) enhanced the expression of tyrosinase mRNA transcripts, tyrosinase synthesis and activity. The increase in tyrosinase activity paralleled the change in the amount of tyrosinase, suggesting that the primary action of alpha-MSH is to stimulate new synthesis of the enzyme. This induction of tyrosinase was associated with the growth of hair that was darker in colour than that of the controls and contained an increased proportion of eumelanin. This increase in eumelanin reflected a decrease in phaeomelanin content. It was concluded that, through its actions on the enzyme tyrosinase, alpha-MSH is able to switch the synthesis of phaeomelanin to that of eumelanin in hair follicular melanocytes.

Tyrosinase and the regulation of coat color changes in C3H-HeAvy mice.

Hair follicular tyrosinase activity was measured during hair growth in neonatal, pubertal, and adult C3H-HeAvy mice that show differences in coat color as a result of changes in the synthesis of eumelanin and pheomelanin. Tyrosinase activity increased during hair growth in all mice but higher levels were found at puberty, when the mice grow a dark, eumelanin coat of hair, than during early and adult life, when the hair follicular melanocytes produce mainly pheomelanin. This suggests that tyrosinase is more important for the synthesis of eumelanin than that of pheomelanin. The increased tyrosinase activity associated with eumelanogenesis in the pubertal mice could not be explained by enhanced transcription of the tyrosinase gene or enzyme synthesis and appeared to be the result of a post-translational activation. Such an activation of tyrosinase was lacking in the neonatal and adult mice; in the latter this was accompanied by a reduction in the glycosylation of tyrosinase and the proportion of enzyme associated with the melanosomal fraction. Our findings suggest that post-translational mechanisms are important control points in the regulation of tyrosinase and that differences in their level of activation are responsible for determining the patterns of melanogenesis in the C3H-HeAvy mice, but it is still not clear how these mechanisms are regulated. Although cyclic AMP increased tyrosinase synthesis it had no post-translation activating effect. The neonatal mice, unlike their pubertal and adult counterparts, also lacked dopachrome converting activity and TRP tyrosinase-related protein-1 expression.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of tyrosinase in hair follicular melanocytes of the mouse during the synthesis of eumelanin and phaeomelanin.

Tyrosinase activity, synthesis, mRNA, and its posttranslational processing were compared in hair follicular melanocytes of the C3HHeAvy mouse during eumelanogenesis and phaeomelanogenesis. Tyrosinase activity was increased during eumelanogenesis; this increase was accompanied by an increase in tyrosinase synthesis. Tyrosinase activity was also increased during phaeomelanogenesis, but only to a peak level that was 50% of that during eumelanogenesis. However, tyrosinase synthesis and mRNA levels were the same in follicles during eumelanin and phaeomelanin synthesis. The lower level of tyrosinase activity is, therefore, presumably due to posttranslational regulation. Less tyrosinase was associated with the particulate fraction during phaeomelanogenesis than during eumelanogenesis. Glycosylation of tyrosinase during phaeomelanogenesis was also reduced and may be the mechanism of control. Bromo-adenosine 3,5-cyclic monophosphate sodium salt increased glycosylation in both eumelanin and phaeomelanin, producing follicles; but this did not result in an increased uptake of tyrosinase onto the melanosome membrane. Therefore, although cAMP increased glycosylation of tyrosinase, the uptake of tyrosinase by the melanosome membrane appeared to be regulated by other systems that are limiting during phaeomelanogenesis, resulting in a lower level of tyrosinase activity.

Effects of γ-radiation on the differentiation of mouse melanocytes in the hair follicles

Pregnant mice were whole-body irradiated with a single acute dose of γ-rays ( 60Co) to investigate the effect of γ-radiation on embryonic melanoblasts. The effect was studied by scoring changes in the differentiation of melanocytes in the hair follicles of mice heterozygous for the recessive coat color mutation pink-eyed dilution ( p). Abnormal round melanocytes were found in the hair matrix and the dermal papilla of F 1 offspring 3.5 days after birth. However, these round melanocytes possessed a melanin deposition of similar intensity to normal hair follicular melanocytes. The frequency of the abnormal hair follicles increased in a dose-dependent manner. Moreover, higher frequencies were found in the animals irradiated at earlier stages of embryonic development. These results indicate that γ-radiation affects dendritogenesis and the location of mouse melanocytes in the hair follicles, with greater effects seen at the earlier stages of development.

Regulation of Tyrosinase Synthesis and its Processing in the Hair Follicular Melanocytes of the Mouse During Eumelanogenesis and Phaeomelanogenesis

Tyrosinase synthesis and its posttranslational processing was compared in hair follicular melanocytes of C3H-HeAvy mice during eumelanogenesis and phaeomelanogenesis. Tyrosinase activity was increased during eumelanogenesis and this was paralleled by an increase in tyrosinase synthesis, as measured by the incorporation of [35S]methionine. Although tyrosinase activity was lower during phaeomelanogenesis there was no change in tyrosinase synthesis. alpha-Melanocyte stimulating hormone (alpha-MSH) increased tyrosinase activity and its synthesis during eumelanogenesis but not during phaeomelanogenesis. Bromo-adenosine 3,5-cyclic monophosphate sodium salt (8-bromo-cAMP) was similarly effective during eumelanogenesis, but unlike alpha-MSH stimulated tyrosinase synthesis during phaeomelanogenesis. This suggests that during phaeomelanogenesis the melanocytes may fail to express MSH receptors. This cannot account for the lower tyrosinase activity, however, for alpha-MSH acts predominantly at the level of tyrosinase synthesis and this was similar during eumelanin and phaeomelanin production. The reduced tyrosinase activity is, therefore, presumably due to some posttranslational change. Accordingly, less tyrosinase was associated with the melanosomal fraction during phaeomelanogenesis than during eumelanogenesis. Glycosylation of tyrosinase, as measured by the incorporation of [3H]glucosamine was also reduced during phaeomelanogenesis. Although 8-bromo-cAMP increased glycosylation of tyrosinase in both eumelanin and phaeomelanin producing melanocytes, it failed to alter the subcellular distribution of the enzyme. It would appear that, although glycosylation of tyrosinase is lower during phaeomelanogenesis, the reduced tyrosinase expression is the result of decreased uptake of tyrosinase by the melanosome.

Effect of alpha-melanocyte-stimulating hormone on tyrosinase activity in hair follicular and epidermal melanocytes of the mouse.

In this study, the effect of alpha-MSH on tyrosinase activity was compared in epidermal and hair follicular melanocytes of mice. It had no effect on epidermal tyrosinase activity in dorsal skin from neonatal non-agouti black mice (C57BL/6J) in both in-vivo and in-vitro experiments. Theophylline and 8-bromocyclic (c)AMP were similarly without effect in in-vitro experiments. In-vivo administration of alpha-MSH and theophylline for 7 days was also without effect on epidermal tyrosinase activity in ear skin of adult non-agouti mice, and the same was true for alpha-MSH in wild-type agouti mice. Activation of the epidermal melanocytes in the non-agouti and wild-type agouti mice with ultraviolet radiation also failed to bring about a response to alpha-MSH and to theophylline in the case of the former. No tyrosinase activity was detected in the epidermis of viable yellow mice (C3H-HeAvy), but, as shown previously, tyrosinase activity was present in the hair follicle when the hair was actively growing and was increased in those mice given either alpha-MSH or theophylline. alpha-MSH and theophylline had no such effects on hair follicular tyrosinase activity in the non-agouti mice. The present results suggest that alpha-MSH- and cAMP-dependent mechanisms have little or no importance in the regulation of tyrosinase expression in mouse epidermal melanocytes. alpha-MSH may, however, regulate tyrosinase expression in hair follicular melanocytes, but even in these melanocytes its action may be restricted to mice that express the agouti gene.

Regulation of tyrosinase synthesis by alpha-melanocyte-stimulating hormone in hair follicular melanocytes of the mouse.

Tyrosinase activity was increased in hair follicular melanocytes of C3H-HeAvy mice during the hair cycle and reached higher levels on days 6-8 after plucking than on day 12. Similarly, the rate of incorporation of [35S]methionine into tyrosinase was greater on days 6-8 than on day 12, but the relative difference was much less. alpha-MSH had no effect on tyrosinase activity or the rate of [35S]methionine incorporation on day 12 and, while it increased both on days 6 and 8, it had a greater effect upon the latter. Pulse-chase experiments showed that the half-life of tyrosinase was 3.5 h and that this was unaffected by alpha-MSH. The results indicate that the increases in tyrosinase activity which occur during the hair cycle involve changes in both the synthesis and activation of the enzyme and that the predominant effect of alpha-MSH is on the former of these two processes.

Dopaminergic inhibition of tyrosinase activity in hair follicular melanocytes of the mouse.

Bromocriptine, a dopamine agonist that blocks the secretion of MSH, inhibits melanogenesis in the hair follicular melanocytes of pubertal C3H-HeAvy mice. However, since this effect cannot be explained by a reduction in circulating alpha-MSH, we have examined the possibility that dopaminergic mechanisms may have a direct inhibitory effect on these melanocytes. Bromocriptine decreased tyrosinase activity in skin explants from 30- to 35-day-old mice that were growing dark hair. This decrease in tyrosinase activity was blocked by dopamine receptor antagonists, haloperidol or spiperone. The specific D2 agonist LY 171555 also inhibited tyrosinase activity in the skin explants in a dose-related manner and the effect was blocked by sulpiride, a D2-receptor antagonist. Neither bromocriptine nor LY 171555 had any effect on tyrosinase activity in skin explants taken from adult mice that were growing yellow hair. The D1-receptor agonist SKF 38393 had no effect on tyrosinase activity in skin explants from either group of mice. The present results support the idea that dopamine D2-receptor agonists have a direct inhibitory effect upon tyrosinase activity of hair follicular melanocytes of the C3H-HeAvy mouse. However, this effect was confined to periods of dark hair growth when the melanocytes produce eumelanin. The D2 agonists were ineffective in reducing tyrosinase activity during adult life when the melanocytes produce predominantly phaeomelanin. This suggests that different control mechanisms may operate in the hair follicular melanocytes during periods of eumelanin and phaeomelanin synthesis.

Melanocyte-stimulating hormone and the regulation of tyrosinase activity in hair follicular melanocytes of the mouse.

Skin tyrosinase activity increases during hair growth in C3H-HeAvy mice and reaches higher levels in young (30- to 35-day-old) mice when the hair follicular melanocytes synthesize the black pigment, eumelanin, than in older (6-month-old) mice when they produce the golden yellow pigment, phaeomelanin. To examine the regulation of the melanocytes at these different stages we have compared the effect of alpha-MSH and other agents that act, through cyclic AMP-dependent mechanisms, on skin tyrosinase activity in both young and old mice during hair growth, initiated by plucking. Daily administration of alpha-MSH, isoprenaline or theophylline increased coat darkness, and skin tyrosinase activity in the younger mice 7-9 days after plucking, but they were ineffective in the older mice. Similarly alpha-MSH, 8-bromo-cyclic AMP or theophylline increased tyrosinase activity in skin explants from the younger mice incubated for up to 24 h but had no effect in explants from older mice. Cyclic GMP had no effect on tyrosinase activity in skin explants from both young and old mice. It is suggested that whereas cyclic AMP-dependent mechanisms may operate to regulate tyrosinase activity in the hair follicular melanocytes of younger mice that produce eumelanin these systems may not operate in the older mice when these melanocytes synthesize phaeomelanin. Phaeomelanin synthesis, unlike that of eumelanin, may not depend upon tyrosinase and its regulation by cyclic AMP and this could explain the low levels of this enzyme in the skin and its failure to respond to alpha-MSH and other activators of the cyclic AMP system during periods of phaeomelanin production.

Melanocyte-stimulating hormone, tyrosinase activity and the regulation of eumelanogenesis and phaeomelanogenesis in the hair follicular melanocytes of the mouse.

Skin tyrosinase levels and the eumelanin and phaeomelanin contents of the hair were measured in pubertal and adult C3H-HeA*vy mice that grow dark and golden yellow hair respectively. Hair growth was initiated by plucking and the skin tyrosinase levels, which increased during the growth of new hair and peaked at around 9 days after plucking, were higher during the growth of dark hair in the pubertal mice than during the growth of yellow hair in the adult mice. Although there was only a twofold difference in the phaeomelanin contents of these two types of hair, the dark hair of the pubertal mice contained over 20 times more eumelanin than the golden-yellow hair of the adult mice. These results suggest that the changes in coat colour in C3H-HeA*vy mice are due mainly to changes in eumelanin synthesis by the hair follicular melanocytes and that the production of this pigment requires higher levels of the enzyme tyrosinase than does the production of phaeomelanin. These changes did not appear to be related to plasma alpha-MSH levels. Nevertheless, administration of alpha-MSH increased skin tyrosinase activity in the pubertal mice that were growing dark hair and produced a twofold increase in the eumelanin content of the hair. However, it had no such effects in adult mice and also failed to affect the phaeomelanin content of the hair in both groups of mice. In contrast to alpha-MSH, bromocriptine decreased skin tyrosinase levels and the eumelanin content and increased the phaeomelanin content of the hair in pubertal mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Histogenesis of the intradermal melanocytic tumor in BDF1 mice induced by topical application of 9,10-dimethyl-1,2-benzanthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA).

The histogenesis of the intradermal melanocytic tumor induced in the skin of 88 female BDF1 mice by DMBA and TPA is studied light-and electron-microscopically. Four types of melanocytes were found in the mouse skin. The epidermal melanocytes transiently appeared several days after topical application of DMBA. Hair follicles turned into anagen phase and follicular melanocytes became apparent by week 3. After week 3, intradermal melanocytes of the perifollicular melanocytic networks (PFM), which were scattered in untreated mouse skin, proliferated to form the intradermal melanocytic tumors, and 267 tumors, composed of oval melanocytes in medullary growth, were induced in all of the 19 treated mice by week 32. Large dendritic melanocytes located deep in the reticular dermis, which did not participate in the formation of the tumor. Only 3 papillomas were induced in 2 of the 19. Light-and electron-microscopically, there was no evidence of migration of epidermal and hair follicular melanocytes into the dermis. Additionally, melanin-producing activity in the cytoplasm of Schwann cell and perineural epithelium was observed. The histogenesis of the intradermal melanocytic tumor was closely related to the melanocytes of the perifollicular melanocytic network (PFM).