Short Report Polymorphisms Upstream of the Melanocortin-1 Receptor Coding Region are Associated with Human Pigmentation Variation in a Brazilian Population VANESSA NEITZKE-MONTINELLI, 1 TURAN P. URMENYI, 1 EDSON RONDINELLI, 1,2,3 PEDRO HERNAN CABELLO, 4 ROSANE SILVA, 1,2 AND RODRIGO S. MOURA-NETO 5,6 * 1 Instituto de Biofı´sica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil 2 Instituto Nacional para Pesquisa Translacional em Sau ´ de e Ambiente na Regia ˜ o Amazo ˆnica, Conselho Nacional de Desenvolvimento Cientı´fico eTecnolo ´gico/MCT, Brazil 3 Departamento de Clı ´nica Me ´dica, Faculdade de Medicina, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil 4 Fundac ¸a ˜ o Oswaldo Cruz, Instituto Oswaldo Cruz, Rio de Janeiro, RJ 21040-900, Brazil 5 Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil 6 DIPRO, Instituto Nacional de Metrologia Qualidade e Tecnologia, Duque de Caxias, RJ 25250-020, Brazil Objective: We describe an association of two SNPs, rs3212345:C>T and rs3212346:G>A, located approximately 2.5 kb upstream of the melanocortin-1 receptor (MC1R) translation initiation codon, with pigmentation phenotype varia- tion in a Southeast Brazilian miscegenated population. Methods: One hundred thirty-eight genetically unrelated subjects, with multicolor phenotype, were selected from the southeast region of Brazil. Skin, hair and eye color, and tanning ability were rated. Genotypes for each SNP (rs3212345:C>T and rs3212346:G>A) were determined. A logistic regression analysis was performed with the additive model to determine which of the polymorphisms contributed to a specific phenotype. Results: We found that the rs3212345:C>T is associated with light skin, red hair, and poor tanning ability, while the rs3212346:G>A is associated with dark skin, black hair, and strong tanning ability. The presence of rs3212345-C and rs3212346-A alleles in human, chimpanzee, gorilla, orangutan, and marmoset genomes suggests that they are the ancestral alleles. Conclusion: These data suggest that the rs3212345-T and rs3212346-G alleles may have contributed to lighter pigmentation phenotypes in modern humans. Genotyping for these SNPs may prove useful to the fields of molecular anthropology and forensic genetics. Am. J. Hum. Biol. 00:000–000, 2012. ' 2012 Wiley Periodicals, Inc. BACKGROUND The melanocortin-1 receptor (MC1R) belongs to the family of G protein-coupled receptors (GPCR) known as the melanocortin receptors (MCRs). The MC1R gene is located on chromosome 16q24.3 and has a single exon of 951 bp that encodes a protein of 317 amino acids. This re- ceptor is considered one of the key proteins in the modula- tion of melanin biosynthesis. Human melanogenesis involves the production of two melanin pigments that are derived from tyrosine. The most abundant is eumelanin, a dark pigment that is found in higher concentrations in darker hair and skin types. Pheomelanin is a red pigment, the concentration of which varies considerably between individuals within any given human group. Higher con- centrations of pheomelanin are often found among red- haired northern Europeans, as well in East Asian and Native Americans (Jablonski, 2004). Certain single nucleotide polymorphisms (SNPs) have been associated with different pigmentation phenotypes within a variety of isolated populations (Kayser and Schneider, 2009). The genetically admixed Brazilian popu- lation is derived from three parental populations: with phenotypical variation occurring by geographical region (Pimenta et al., 2006; Raimann et al., 2012). This work investigates the association between two SNPs upstream of the MC1R gene promoter region with pigmentation phenotype variation in a Southeast Brazilian miscegen- ated population. Such investigation can provide valuable information about phenotypic features from human skele- tal remains, for molecular anthropologists, and forensic geneticists (Bouakaze et al., 2009; Walsh et al., 2011). METHODS Population sample Subjects with multicolor phenotype, as defined by National Survey Inquiries of Brazilian Government, were selected from the southeast region of Brazil. We ascer- tained 138 genetically unrelated individuals and had clas- sified the pigmentation phenotype. Skin color was rated on a three-category scale (light, intermediate, and dark), hair color on a four-category scale (red, light, brown, and black), and eye color on a three-category scale (blue, green/hazel, and dark). Tanning ability was classified according to a modified Fitzpatrick scale (Fitzpatrick, 1975). Type I (no tanning ability) was not present in the sample due to the paucity of this complexion among Bra- zilians. Therefore, tanning ability was categorized in three types: Type II (poor tanning ability with high sensi- tivity to the sun), Type III (medium tanning ability and medium sensitivity to the sun), and Type IV (high tanning ability and insensitivity to the sun). Buccal cell DNA Additional Supporting Information may be found in the online version of this article. Contract grant sponsors: CAPES, CNPq; Contract grant sponsor: FAPERJ; Contract grant number: E26/111.537/2010. *Correspondence to: Dr. Rodrigo S. Moura-Neto, Laborato ´rio de Biologia Molecular Forense, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho, 373, Ed. CCS, Bloco A, Sala A0-087, Rio de Janeiro, RJ – CEP 21.941-902, Brasil. E-mail: rodrigomouraneto@ufrj.br Received 22 May 2012; Revision received 13 June 2012; Accepted 19 June 2012 DOI 10.1002/ajhb.22301 Published online in Wiley Online Library (wileyonlinelibrary. com). AMERICAN JOURNAL OF HUMAN BIOLOGY 00:000–000 (2012) V V C 2012 Wiley Periodicals, Inc.