肤色、黑色素皮质素受体1和紫外线

近期的研究表明,哺乳动物黑素细胞中黑色素皮质素受体1（MC1R）对调节棕黑色素和红黄色素的合成起关键的作用。MC1R基因的变异与动物的皮毛、人的皮肤和头发颜色差异密切相关。对小鼠的遗传学研究显示,MC1R是独特的、双功能控制受体。它由α-促黑色素皮质激素激活,其拮抗物为agouti蛋白,二者的共同作用导致哺乳动物表皮颜色的变异。另外,人类皮肤的色素沉着是决定于皮肤对外辐射的反应,以及由此引发皮肤癌的重要因素。MC1R变异与黑色素癌易感性相关。 Genotype,Melanocortin 1 Receptor and Ultrviolet Radiation Lü Xue-mei,SHI Peng,ZHANG Ya-ping Lab of Cellular & Molecular Evolution,Kunming Institute of Zoology,the Chinese Academy of Sciences,Kunming 650223,China Abstract:Recent work on the melanocortin 1 receptor (MC1R) suggests that MC1R plays a central role in regulation of eumelanin (brwon/black melanins) and phaeomelanin (red/yellow melanins) synthesis within the mammalian melanocyte.In the mouse,genetic studies show that the MC1R appears to be a unique,bifunctionally controlled receptor,activated by α–MSH and antagonized by agouti,both of which contribute to the variability seen in mammalian coat color.Variants of this receptor are associated with different animal's coat,human skin and hair colors.In addition,cutaneous pigmentation is a major determinant of the cutaneous response to ultraviolet radiation,and consequently of the risk of developing skin cancer.MC1R variants are a risk factor for melanoma susceptibility. Key words：Melanocortin-1 receptor gene; MC1R variants; ultraviolet radiation; skin and hair colors; skin cancer     

Ultrastructural change of melanosomes associated with agouti pattern formation in mouse hair.

We have studied the structural alteration of melanosomes in the melanocytes of agouti mice whose genetic characteristic is to produce eumelanin and phaeomelanin alternately in a single hair bulb. Melanocytes of hair bulbs from 1 to 2 day old mice of the black phase were observed to contain rod-shaped melanosomes of the eumelanin type (eumelanosome). In the melanocytes of the hair bulbs from 4 to 6-day old skin, which exclusively contain phaeomelanin, spherical melanosomes (phaeomelanosomes) were seen. On the other hand, the mice of the transitional phase from black to yellow possessed melanocytes that contained both eumelanosomes and phaeomelanosomes within a single cell. This result indicates that the shift from the eumelanin formation to the phaeomelanin formation or vice versa in agouti hair occurs within a single melanocyte.We observed multivesicular bodies in both the agouti melanocytes of the yellow phase and the genotypically yellow melanocytes. These bodies are considered to be the precursor of the phaeomelanin-containing melanosome. They are sometimes observed to have continuity with E. R. suggesting that the melanosomes are derived from E. R. in the phaeomelanin-forming melanocytes.     

Agouti: from mouse to man, from skin to fat

The agouti protein regulates pigmentation in the mouse hair follicle producing a black hair with a subapical yellow band. Its effect on pigmentation is achieved by antagonizing the binding of alpha-melanocyte stimulating hormone (alpha-MSH) to melanocortin 1 receptor (Mc1r), switching melanin synthesis from eumelanin (black/brown) to phaeomelanin (red/yellow). Dominant mutations in the non-coding region of mouse agouti cause yellow coat colour and ectopic expression also results in obesity, type 11 diabetes, increased somatic growth and tumourigenesis. At least some of these pleiotropic effects can be explained by antagonism of other members of the melanocortin receptor family by agouti protein. The yellow coat colour is the result of agouti chronically antagonizing the binding of alpha-MSH to Mc1r and the obese phenotype results from agouti protein antagonizing the binding of alpha-MSH to Mc3r and/or Mc4r. Despite the existence of a highly homologous agouti protein in humans, agouti signal protein (ASIP), its role has yet to be defined. However it is known that human ASIP is expressed at highest levels in adipose tissue where it may antagonize one of the melanocortin receptors. The conserved nature of the agouti protein combined with the diverse phenotypic effects of agouti mutations in mouse and the different expression patterns of human and mouse agouti, suggest ASIP may play a role in human energy homeostasis and possibly human pigmentation.     

Female Fertility and Mating Type Effects on Effective Population Size and Evolution in Filamentous Fungi

The murine agouti locus regulates a switch in pigment synthesis between eumelanin (black/brown pigment) and phaeomelanin (yellow/red pigment) by hair bulb melanocytes. We recently described a spontaneous mutation, hypervariable yellow (A(hvy)) and demonstrated that A(hvy) is responsible for the largest range of phenotypes yet identified at the agouti locus, producing mice that are obese with yellow coats to mice that are of normal weight with black coats. Here, we show that agouti expression is altered both temporally and spatially in A(hvy) mutants. Agouti expression levels are positively correlated with the degree of yellow pigmentation in individual A(hvy) mice, consistent with results from other dominant yellow agouti mutations. Sequencing of 5' RACE and genomic PCR products revealed that A(hvy) resulted from the integration of an intracisternal A particle (IAP) in an antisense orientation within the 5' untranslated agouti exon 1C. This retrovirus-like element is responsible for deregulating agouti expression in A(hvy) mice; agouti expression is correlated with the methylation state of CpG residues in the IAP long terminal repeat as well as in host genomic DNA. In addition, the data suggest that the variable phenotype of A(hvy) offspring is influenced in part by the phenotype of their A(hvy) female parent.     

The genetics of coat colors in the mongolian gerbil (Meriones unguiculatus)

Genetic studies demonstrated three loci controlling coat colors in the Mongolian gerbil. F1 hybrids of white gerbils with red eyes and agouti gerbils with wild coat color had the agouti coat color. The segregating ratio of agouti and white in the F2 generation was 3:1. In the backcross (BC) generation (white x F1), the ratio of the agouti and white coat colors was 1:1. Next, inheritance of the agouti coat color was investigated. Matings between agouti and non-agouti (black) gerbils produced only agouti gerbils. In the F2 generation, the ratio of agouti to non-agouti (black) was 3:1. There was no distortion in the sex ratios within each coat color in the F1, F2 and BC generations. This indicated that the white coat color of gerbils is governed by an autosomal recessive gene which should be named the c allele of the c (albino) locus controlling pigmentation, and the agouti coat color is controlled by an autosomal dominant gene which might be named the A allele of the A (agouti) locus controlling pigmentation patterns in the hair. The occurrence of the black gerbil demonstrated clearly the existence of the b (brown) locus, and it clearly indicated that the coat colors of gerbils can basically be explained by a, b, and c loci as in mice and rats.     

Molecular Genetic Characterization of Six Recessive Viable Alleles of the Mouse Agouti Locus

The agouti locus on mouse chromosome 2 encodes a secreted cysteine-rich protein of 131 amino acids that acts as a molecular switch to instruct the melanocyte to make either yellow pigment (phaeomelanin) or black pigment (eumelanin). Mutations that up-regulate agouti expression are dominant to those causing decreased expression and result in yellow coat color. Other associated effects are obesity, diabetes, and increased susceptibility to tumors. To try to define important functional domains of the agouti protein, we have analyzed the molecular defects present in a series of recessive viable agouti mutations. In total, six alleles (a(mJ), a(u), a(da), a(16H), a(18H), a(e)) were examined at both the RNA and DNA level. Two of the alleles, a(16H) and a(e), result from mutations in the agouti coding region. Four alleles (a(mJ), a(u), a(18H), and a(da)) appear to represent regulatory mutations that down-regulate agouti expression. Interestingly, one of these mutations, a(18H), also appears to cause an immunological defect in the homozygous condition. This immunological defect is somewhat analogous to that observed in motheaten (me) mutant mice. Short and long-range restriction enzyme analyses of homozygous a(18H) DNA are consistent with the hypothesis that a(18H) results from a paracentric inversion where one end of the inversion maps in the 5' regulatory region of agouti and the other end in or near a gene that is required for normal immunological function. Cloning the breakpoints of this putative inversion should allow us to identify the gene that confers this interesting immunological disorder.     

Pituitary hypoplasia in Pttg-/- mice is protective for Rb+/- pituitary tumorigenesis

Pituitary tumor transforming gene (Pttg) is induced in pituitary tumors and associated with increased tumor invasiveness. Pttg-null mice do not develop tumors, but exhibit pituitary hypoplasia, whereas mice heterozygous for the retinoblastoma (Rb) deletion develop pituitary tumors with high penetrance. Pttg-null mice were therefore cross-bred with Rb+/- mice to test the impact of pituitary hypoplasia on tumor development. Before tumor development, Rb+/-Pttg-/- mice have smaller pituitary glands with fewer cycling pituitary cells and exhibit induction of pituitary p21 levels. Pttg silencing in vitro with specific short hairpin interfering RNA in AtT20 mouse corticotrophs led to a marked induction of p21 mRNA and protein levels, decreased RB phosphorylation, and subsequent 24% decrease in S-phase cells. Eighty-six percent of Rb+/-Pttg+/+ mice develop pituitary adenomas by 13 months, in contrast to 30% of double-crossed Rb+/-Pttg-/- animals (P < 0.01). Pituitary hypoplasia, associated with suppressed cell proliferation, prevents the high penetrance of pituitary tumors in Rb+/- animals, and is therefore a protective determinant for pituitary tumorigenesis.     

The mouse pink-eyed dilution allele of the P-gene greatly inhibits eumelanin but not pheomelanin synthesis

The mouse pink-eyed dilution (p) locus is known to control eumelanin synthesis, melanosome morphology, and tyrosinase activity in melanocytes. However, it has not been fully determined whether the mutant allele, p affects pheomelanin synthesis. Effects of the p allele on eumelanin and phemelanin synthesis were investigated by chemical analysis of dorsal hairs of 5-week-old mice obtained from the F(2) generations (black, pink-eyed black, recessive yellow, pink-eyed recessive yellow, agouti, and pink-eyed agouti) between C57BL/10JHir (B10)-congenic pink-eyed black mice (B10-p/p) and recessive yellow (B10-Mc1r(e)/Mc1r(e)) or agouti (B10-A/A) mice. The eumelanin content was dramatically (>20-fold) decreased in pink-eyed black and pink-eyed agouti mice, whereas the pheomelanin content did not decrease in pink-eyed black, pink-eyed recessive yellow, or pink-eyed agouti mice compared to the corresponding P/- mice. These results suggest that the pink-eyed dilution allele greatly inhibits eumelanin synthesis, but not pheomelanin synthesis.     

The effect of methyl-containing supplements during pregnancy on the phenotypic modification of offspring hair color in rats

The effect of methyl supplements to the diet of pregnant homozygous (AAHH) female rats with agouti coat color mated with homozygous (aahh) males on the phenotypic modification of the coat color of their heterozygous offspring (AaHh) has been studied. Comparative morphological analysis of the main parameters of hair that determine coat color, including the total length of hairs of different types and the length of the upper black (eumelanin) and light (pheomelanin) parts of awn hairs has been performed. The pattern of pigment granule distribution among hair layers has been analyzed. The melanin content of the hair has been determined using electron spin resonance (ESR). Although all offspring have a typical agouti coat color (alternating black and light portions of hair), 39% of them have a darker coat color than control and other experimental rats have. The main differences between the offspring with darkened and standard coat colors are accounted for by the ratio between the eumelanin and pheomelanin portions of awn hairs. In darkened offspring, this ratio is significantly higher than in control rats. The possible mechanisms of the phenotypic modification of agouti coat color in experimental animals are discussed.     

The effect of methyl supplements during pregnancy on the phenotypic modification of offspring agouti coat color in rats

The effect of methyl supplements to the diet of pregnant homozygous (AAHH) female rats with agouti coat color mated with homozygous (aahh) males on the phenotypic modification of the coat color of their heterozygous offspring (AaHh) has been studied. Comparative morphological analysis of the main parameters of hair that determine coat color, including the total length of hairs of different types and the length of the upper black (eumelanin) and light (pheomelanin) parts of awn hairs has been performed. The pattern of pigment granule distribution among hair layers has been analyzed. The melanin content of the hair has been determined using electron spin resonance (ESR). Although all offspring have a typical agouti coat color (alternating black and light portions of hair), 39% of them have a darker coat color than control and other experimental rats have. The main differences between the offspring with darkened and standard coat colors are accounted for by the ratio between the eumelanin and pheomelanin portions of awn hairs. In darkened offspring, this ratio is significantly higher than in control rats. The possible mechanisms of the phenotypic modification of agouti coat color in experimental animals are discussed.     

Fox colors in relation to colors in mice and sheep

Color inheritance in foxes is explained in terms of homology between color loci in foxes, mice, and sheep. The hypothesis presented suggests that the loci A (agouti), B (black/chocolate brown pigment) and E (extension of eumelanin vs. phaeomelanin) all occur in foxes, both the red fox, Vulpes vulpes, and the arctic fox, Alopex lagopus. Two alleles are postulated at each locus in each species. At the A locus, the (top) dominant allele in the red fox, Ar, produces red color and the corresponding allele in the arctic fox, Aw, produces the winter-white color. The bottom recessive allele in both species is a, which results in the black color of the silver fox and a rare black color in the Icelandic arctic fox when homozygous. The B alleles are assumed to be similar in both species: B, dominant, producing black eumelanin, and b, recessive, producing chocolate brown eumelanin when homozygous. The recessive E allele at the E locus in homozygous form has no effect on the phenotype determined by alleles at the A locus, while Ed, the dominant allele is epistatic to the A alleles and results in Alaska black in the red fox and the dark phase in the arctic fox. Genetic formulae of various color forms of red and arctic fox and their hybrids are presented.     

Maternal methyl supplements affect the phenotypic variation of the <Emphasis Type="Italic">agouti</Emphasis> gene in the offspring of rats with different behavioral types

The effects of selection of agouti rats (with genotype AAHH) on the tame and aggressive behavior and dietary methyl given to females from the eighth day of pregnancy to the fifth day after the birth of the offspring on the intensity of the agouti coat color in the offspring have been studied. The morphometric parameters of hair determining the darkness of the agouti color (the total length of guard hairs, the lengths of their eumelanin end and pheomelanin band, the ratio between the lengths of the eumelanin and pheomelanin portions of the hair, the total length of the awn hairs, and the relative length of their widened “lanceolate” upper end) have been compared. It has been found that selection of agouti rats for aggressive behavior is accompanied by darkening of the coat color compared to tame rats due to an increase in the ratio of the length of the black eumelanin end of the guard hairs to the length of the yellow pheomelanin band. Methyl-containing additives to the diet of females affect the intensity of the agouti coat color in the offsprings with both types of behavior, but to different extents. Aggressive offspring is more sensitive to the mother’s methyl-containing diet: the percentage of animals that are darker than control rats is higher among aggressive animals than among tame ones due to a greater increase in the ratio between dark and light portions of hairs. The possible mechanisms of differences in the phenotypic modifications of coat color in control and experimental agouti rats with different types of behavior are discussed.     

Pigmentation in Black-boned sheep (Ovis aries): association with polymorphism of the MC1R gene

Variations in vertebrate skin and hair color are due to varied amounts of eumelanin (brown/black) and phaeomelanin (red/yellow) produced by the melanocytes. The melanocortin 1 receptor (MC1R) is a regulator of eumelanin and phaeomelanin production in the melanocytes, and MC1R mutations causing coat color changes are known in many vertebrates. We have sequenced the entire coding region of the MC1R gene in Black-boned, Nanping indigenous and Romney Marsh sheep populations and found two silent mutation sites of A12G and G144C, respectively. PCR-RFLP of G144C showed that frequency of allele G in Black-boned, Nanping indigenous and Romney Marsh sheep was 0.818, 0.894 and 0, respectively. Sheep with GG genotype had significantly higher (P < 0.05) tyrosinase activity than sheep with CC genotype in the all investigated samples. Moreover, there was significant effect of MC1R genotype on coat color, suggesting that MC1R gene could affect coat color but not black traits. There would be merit in further studies using molecular techniques to elucidate the cause of black traits in these Black-boned sheep.     

Allelic Variation within the Emv-15 Locus Defines Genomic Sequences Closely Linked to the agouti Locus on Mouse Chromosome 2

Gene(s) at the agouti locus act within the microenvironment of the hair follicle to switch pigment synthesis in the melanocyte between eumelanin (black or brown pigment) and phaeomelanin (yellow pigment). Many phenotypic variants of this locus have been described. The mechanism(s) of gene action causing such variation in coat-color phenotype is not known. The close linkage of an endogenous ecotropic murine leukemia provirus, Emv-15, to the lethal yellow mutation of the agouti locus provides a means to molecularly access genes at or near the agouti locus. We have identified and used a unique mouse sequence flanking the Emv-15 provirus to define three alleles of the Emv-15 locus. We found a correlation between the presence of specific Emv-15 alleles and the origins of specific agouti locus mutations, confirming close linkage. However, we found some exceptions which suggest that the Emv-15 locus is closely linked to, but genetically separable from, the agouti locus.     

Human hair melanins: what we have learned and have not learned from mouse coat color pigmentation

Hair pigmentation is one of the most conspicuous phenotypes in humans. Melanocytes produce two distinct types of melanin pigment: brown to black, indolic eumelanin and yellow to reddish brown, sulfur‐containing pheomelanin. Biochemically, the precursor tyrosine and the key enzyme tyrosinase and the tyrosinase‐related proteins are involved in eumelanogenesis, while only the additional presence of cysteine is necessary for pheomelanogenesis. Other important proteins involved in melanogenesis include P protein, MATP protein, α‐MSH, agouti signaling protein (ASIP), MC1R (the receptor for MSH and ASIP), and SLC7A11, a cystine transporter. Many studies have examined the effects of loss‐of‐function mutations of those proteins on mouse coat color pigmentation. In contrast, much less is known regarding the effects of mutations of the corresponding proteins on human hair pigmentation except for MC1R polymorphisms that lead to pheomelanogenesis. This perspective will discuss what we have/have not learned from mouse coat color pigmentation, with special emphasis on the significant roles of pH and the level of cysteine in melanosomes in controlling melanogenesis. Based on these data, a hypothesis is proposed to explain the diversity of human hair pigmentation.     

Fluorescence spectrophotometric analysis of melanins in the house mouse

We extracted the yellow melanin (phaeomelanin), black melanin (eumelanin), and mixed type of melanin from dorsal hair of dominant yellow (A ^y /a), non-agouti (a/a), and agouti (A/A) mice, respectively. Spectrophotometric and fluorescence spectrophotometric analysis demonstrated that the yellow melanin was qualitatively distinct from the black melanin and that the agouti hair contained both types of pigment.This work was supported by Grant 244004 from the Ministry of Education. Part of this work was presented at the X International Pigment Cell Conference.     

The role of melanocyte-stimulating hormone (MSH) receptor in bovine coat color determination

The melanocyte-stimulating hormone (MSH) receptor has a major function in the regulation of black (eumelanin) versus red (phaeomelanin) pigment synthesis within melanocytes. We report three alleles of the MSH-receptor gene found in cattle. A point mutation in the dominant allele E ^D gives black coat color, whereas a frameshift mutation, producing a prematurely terminated receptor, in homozygous e/e animals, produces red coat color. The wild-type allele E ⁺ produces a variety of colors, reflecting the possibilities for regulating the normal receptor. Microsatellite analysis, RFLP studies, and coat color information were used to localize the MSH-receptor to bovine Chromosome (Chr) 18.     

Electron spin resonance spectrometrical study of the melanins in the wool of some North European sheep in relation to their color inheritance

Additional peaks that were known on the esr-spectrograms of red human and reddish-brown Karakul hair to be diagnostic traits of phaeomelanin esr-signal also were found on esr-spectrograms of the tan, but not of black or chocolate brown wool from Icelandic sheep. This tan color is thought to depend on the presence of phaeomelanin and is due to the top dominant allele at the A locus. The two methods of distinguishing between eu- and phaeomelanin-dependent brown colors--esr-spectrometrical and genetical--are in agreement for European as well as for Asiatic breeds. Both light and dark brown Soay fleece samples lacked the additional peaks and are interpreted as eumelanin pigmentation.     

In situ localization of agouti signal protein in murine skin using immunohistochemistry with an ASP-specific antibody

Switching between production of eumelanin or pheomelanin in follicular melanocytes is responsible for hair color in mammals; in mice, this switch is controlled by the agouti locus, which encodes agouti signal protein (ASP) through the action of melanocortin receptor 1. To study expression and processing patterns of ASP in the skin and its regulation of pigment production in hair follicles, we have generated a rabbit antibody (termed alphaPEP16) against a synthetic peptide that corresponds to the carboxyl terminus of ASP. The specificity of that antibody was measured by ELISA and was confirmed by Western blot analysis. Using immunohistochemistry, we characterized the expression of ASP in the skin of newborn mice at 3, 6, and 9 days postnatally. Expression in nonagouti (a/a) black mouse skin was negative at all times examined, as expected, and high expression of ASP was observed in 6 day newborn agouti (A/+) and in 6 and 9 day newborn lethal yellow (A(y)/a) mouse skin. In lethal yellow (pheomelanogenic) mice, ASP expression increased day by day as the hair color became more yellow. These expression patterns suggest that ASP is delivered quickly and efficiently to melanocytes and to hair matrix cells in the hair bulbs where it regulates melanin production.     

Circling,deafness, and yellow coat displayed by yellow submarine (ysb) and light coat and circling (lcc) mice with mutations on chromosome 3

We describe here two mouse mutants, yellow submarine (Ysb) and light coat and circling (Lcc). Ysb arose as the result of insertions of a transgene, pAA2, into the genome. Lcc is an independent, radiation-induced mutation. Both mutants are characterized by recessive circling behavior and deafness, associated with a non-segregating, semi-dominant yellow coat color. Complementation tests showed that Ysb and Lcc are allelic. We attribute the yellow coat in Ysb and Lcc mice to the absence of black awl overhairs, increased agouti zigzag underhairs, and the presence of agouti awls with long subapical yellow pigment. Chromosomal mapping and genomic characterization showed the Ysb and Lcc mutations involve complex chromosomal rearrangements in overlapping regions of mouse chromosome 3, A2/A3-B/C and B-E1, respectively. Ysb and Lcc show for the first time, to our knowledge, the presence of genes in the B-C region of chromosome 3 important for balance and hearing and the pigmentation and specification of coat hair.