08/10/2023
THE GENETICS OF THE COLOR SPECTRUM IN OUR PUERTO RICAN PASO FINOS
Paola Matos Ruiz & Xiomara R. Arias | Born to Gait Magazine (Fall 2023 Issue)
The genetics article in the previous magazine issue focused on the genetics of tiger eyes, which is a unique trait of the Pure Puerto Rican Paso Finos. In this article, we plan to continue promoting the genetic diversity of the Pure Puerto Rican Paso Fino breed by exploring the genetics of coat colors and blue eyes. We also plan to illustrate the genetics of coat and eye color through a case-based learning series using a Pure Puerto Rican Paso Fino cremello horse as an example.
Mutations. We hear this word and may have vastly different impressions of what it means. Do you think of ugly mutants, abominable creatures that come from the depth of toxic waste? When we are children, this is what may have come first to our minds. Despite being perceived as atrocious by some, mutations allow the world to be filled with diverse beauty and benefits for species’ survival. Divergent biological aspects create a planet with many new and appealing sights. Our world would not be nearly as beautiful or diverse without the possibility of mutations. Due to mutations, we have the diversity of coats and eye colors we appreciate nowadays in horses.
Red and black are the primary pigments in horses’ base coat colors. The interaction of the genes Melanocortin Receptor 1 (MC1R, abbreviated as E) and Agouti-signaling protein (abbreviated as A) determines the base coat color in horses.1-2 How these genes are co-inherited dictates the final base coat color product.
The MC1R gene plays a role in determining the black and red pigments and is known as “extension” or the red factor locus. Three alleles are known for the base coat color dictated by the MC1R gene: E, e, and ea. Two copies of the recessive alleles (e/e, e/ea or ea /ea ) alleles would produce only horses with a red base coat color (e.g, chestnuts, palominos, red duns) because this genotype masks the expression of other genotypes that could be co-inherited through a genetic mechanism known as epistasis. The Agouti gene determines the distribution of black pigmentation in horses and acts as a modifier to determine bay versus black. From the absence of the Agouti dominant copy (A/a or A/A) and recessive red factor alleles (e/e, e/ea or ea /ea ), we can get black horses if the horse genotype at the MC1R gene is E/E or E/e. In the presence of one or two copies of Agouti (A/a or A/A) and one or two for the MC1R gene (E/e or E/E), the horse may have a bay color phenotype with black pigmentation restricted to the mane, tail, lower legs and ear rims. These areas of black pigmentation are referred to as “points”. In other words, Agouti modifies the black pigmentation on the horse's body, except for the points mentioned above.
While black and red (also known as eumelanin and pheomelanin, respectively) are the only pigment types present in horses, mutations can cause a different distribution of pigments, resulting in our diverse color wheel of horse hair. For example, the cremello, smoky black, smoky cream, and perlino colors are the results of a mutation, primarily affecting the Cream locus9-10.
The Cream (C) locus located in chromosome 21 is where the Cr and N alleles are found.9-10 Cr refers to an allele with a variant in the MATP gene that produces color dilution, whereas N refers to an allele without the variant.9-10 Since horses are diploid, they will inherit an allele from each of their parents. The Cream locus can present a different phenotype for homozygotes (e.g., Cr/Cr) and heterozygotes (Cr/N). For example, if a horse is homozygous for the Cream allele (e.g., Cr/Cr), it means it has two copies of the Cream dilution variant. Depending on the base coat color (e.g., chestnut, bay, or black) the homozygous (Cr/Cr) horse would be expected to be cremello (chestnut base), perlino (bay base), or smoky cream (black base). If the horse is heterozygous, Cr/N, it has one copy of the Cream dilution allele, it would be expected to be either palomino (chestnut base), buckskin (bay base), or smoky black (black base). If the horse is recessive for the absence of the variant at the Cream dilution locus, N/N, its phenotype should show its base coat color.
The presence of two copies of the Cream dilution allele at the C gene locus does not render a horse an albino. Albinism refers to the inability to produce pigment and, therefore, a complete lack of color, which presents itself as white. The presence of Cream does not result in the absence of color; rather, it changes the distribution of pigment. Albinism is currently not a phenomenon that has been described for horses. Although from manifestations in other species, albinism often causes reddish/pink eyes.11 Cremello and perlino horses have blue eyes, therefore demonstrating that they are not albinos.
Pintos, similarly, are a result of a variety of genetic mutations. The patterns of pintos that have been observed in the Puerto Rican Paso Finos are: Tobiano pintos (TO), Sabino pintos (SB1) and Dominant White 20 (DW20). Tobiano pintos are a result of a chromosomal inversion in chromosome #3.8 The specific genetic mutation observed in our breed’s Sabino pintos has not been identified, but it might involve the KIT gene. The Sabino mutations tend to vary among horse breeds; the specific mutation for the Pure Puerto Rican Paso Finos has not been identified scientifically, but we predict it might involve the KIT gene.4 The DW20 results from a point mutation at the KIT gene (c.2045G>A) and results in prominent white markings in the leg and face, more in its homozygous form than its heterozygous form.3 From talking to breeders in Puerto Rico who own offspring of the Grand Sire of the Breed, Linaje D’ La Excelencia, the DW20 mutation has been confirmed genetically through genetics panels ordered by the breeders themselves. Based on pedigree analysis, most likely, Linaje is a carrier of this mutation because of his paternal grandmother, Jackeline.
So far, research has not found evidence about the Tobiano, Sabino, and DW20 mutations resulting in adverse health effects. In contrast, gray horses, which have also been observed in our breed, do experience adverse effects by being at a higher risk of developing melanomas in their lifetime due to depigmentation of their skin. Gray horses are a result of a mutation (i.e. duplication of intron 6) in the gene syntaxin 17 (STX17) that results in progressive depigmentation of the skin by inhibiting the production of melanocytes.5-6 They are born gray and progressively their hair color transforms to white. The pattern of inheritance is autosomal dominant, which implies you only need one copy of the mutated allele of gene STX17 to develop this pattern. In other words, they lose their hair pigmentation which serves as an armor to protect their bodies against the dangerous sun UV light and are prone to develop skin cancer. According to most recent research, 70-80% of gray horses will develop melanoma (skin cancer) by the time they are 15 years old.5-6 It is important to clarify that some of the skin tumors gray horses develop are benign; about 66% become malignant.7
To current knowledge, there are only two Pure Puerto Rican Paso Fino double cream diluted stallions worldwide. They are both cremellos, with Sueño de Paz residing in Puerto Rico and Rayito de Oro residing in the mainland United States. Sueño de Paz sire is the palomino stallion Ensueño Dorado and palomino dam Platanera Dorada. Rayito de Oro is the son of the palomino stallion Cale's Helado Oro and buckskin dam Tostada Hidalgo.
These stallions will always pass on one of their Cream alleles to their offspring. This means that if you mate them to a bay mare, you will (mostly) get buckskins. We say ‘mostly’ because if the bay horse base coat color genotype is heterozygous for the MC1R gene (e.g., E/e), there is a 50% chance the offspring will be a palomino, and if you mate them to a chestnut (e/e), you will get a palomino.
The cremellos of our breed not only contribute to the maintenance and boost of the Pure Puerto Rican Paso Fino coat color spectrum but also eye color diversity. These cremellos have blue eyes primarily due to their genotype at the Cream locus. It is important to highlight that blue eyes have been observed on horses of all coat colors but are more prevalent and more likely to be inherited from double-dilute horses, such as cremellos, and genes that cause white, such as pintos. In our beloved Pure Puerto Rican Paso Finos, blue eyes have been observed in pintos, cremellos, and even bay horses. Interestingly, some of the bay horses with blue eyes have one of the two eyes affected. This phenomenon is called heterochromia.
Sueño de Paz (Ensueño Dorado x Platanera Dorada) is a cremello with blue eyes - Sueño de Paz sire and dam were both palominos. Therefore, he inherited one copy of the Cr from each parent to have a cremello phenotype. Sueño de Paz’s owner, Ms. Viveca Venegas Vilaró, kindly accepted our request for the genetic panels of her stallion and some of his offspring. His genetic panel confirms he is red-based (e/e) and homozygous for the Cream locus (Cr/Cr). He is also homozygous for the Agouti trait, but this trait is masked by the presence of two copies of the red factor (e/e). Given his homozygosity on the Cream locus, he can only produce buckskins or palominos when bred to a bay mare or palominos when bred to a chestnut mare. When bred to a palomino, Sueño de Paz offers a 50% chance of producing a cremello. When bred to a chestnut mare, he produces 100% palominos.
Interestingly, Sueño de Paz is a carrier of the tiger eye allele 2. Therefore, this outstanding cremello Pure Puerto Rican Paso Fino horse not only can produce palominos and buckskins, but can also help increase the number of Pure Puerto Rican Paso Finos with tiger eyes if bred to a carrier or tiger-eye mare. From the data shared by Ms. Venegas Vilaró, Sueño de Paz, has already produced offspring with tiger eyes. Here are some examples of his offspring (genetic panel information was confirmed):
1- Compositora de Sueños (Sueño de Paz x Sara de Plebeyo*) - Palomino mare with tiger eyes.
2- Cantares de Nan (Sueño de Paz x Sara de Plebeyo*) - Buckskin mare with tiger eyes.
*Sara de Plebeyo (Plebeyo x Sarita M.) - Is a bay mare heterozygous for the red factor (Ee). Therefore, she has a chance of producing 50% palominos and 50% buckskins when bred to Sueño de Paz.
The versatility of the Puerto Rican Paso Fino is one of the key features of the breed. While we often focus on what they can do in terms of riding and intelligence, we must not neglect to recognize the brilliant diversity of color that also contributes to the uniqueness of the breed. Preserving color while maintaining the important aspects of the Puerto Rican Paso Fino—gait, conformation, intelligence—is an important task to take accountability for and continue. We must not forget that the tiger eye is one of the defining features of the breed that can genetically show the Puerto Rican Paso Fino is different from other similarly gaited breeds. The tiger eye indicates that our breed has been bred separately from other populations, creating a unique breed in need of conservation.
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PLEASE SEE THE TABLE IN THE ACCOMPANYING PHOTOS
Table 1. Summary of the genotype and corresponding phenotype for some of the colors observed in the Puerto Rican Paso Fino.
Disclaimer: This table is not comprehensive and does not include the different color subtypes described in horses of our breed.
*N= normal/wild-type allele
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Acknowledgments:
We want to thank Viveca Venegas Vilaró, who collaborated with us to make this article possible.
References:
1- Rieder, S., Taourit, S., Mariat, D. et al. Mutations in the agouti (ASIP), the extension (MC1R), and the brown (TYRP1) loci and their association to coat color phenotypes in horses. Equus caballus. 12, 450–455 (2001).
2- UC Davis website (Bay, black and chestnut horses): https://vgl.ucdavis.edu/test/agouti-horse, https://vgl.ucdavis.edu/test/red-factor-horse
3- Book- Horse Genetics, 3rd edition | Drs. Ernest Bailey and Samantha Brooks | Wallingford, Oxfordshire, UK; Boston; CABI [2020].
4- Hauswirth R, Jude R, Haase B, Bellone RR, Archer S, Holl H, Brooks SA, Tozaki T, Penedo MC, Rieder S, Leeb T. Novel variants in the KIT and PAX3 genes in horses with white-spotted coat color phenotypes. Anim Genet. 44(6):763-5 (2013).
5- Curik I, Druml T, Seltenhammer M, Sundström E, Pielberg GR, Andersson L, Sölkner J. Complex inheritance of melanoma and pigmentation of coat and skin in Grey horses. PLoS Genet. 9(2):e1003248 (2013).
6- UC Davis website (Grey Horses): https://vgl.ucdavis.edu/test/gray
7- Yi Z, Gao Y, Yu F, Zhu Y, Liu H, Li J, Murua Escobar H. Interventions for treatment of cutaneous melanoma in horses: a structured literature review. Vet Res Commun. (2022).
8- Brooks SA, Lear TL, Adelson DL, Bailey E. A chromosome inversion near the KIT gene and the Tobiano spotting pattern in horses. Cytogenet Genome Res. 119(3-4):225-30 (2007).
9- Mariat D, Taourit S, Guérin G. A mutation in the MATP gene causes the cream coat color in the horse. Genet Sel Evol. 35(1):119-33 (2003).
10- Locke MM, Ruth LS, Millon LV, Penedo MC, Murray JD, Bowling AT. The cream dilution gene, responsible for the palomino and buckskin coat colors, maps to horse chromosome 21. Anim Genet. 32(6):340-3 (2001).
11- White, D., Rabago-Smith, M. Genotype–phenotype associations and human eye color. J Hum Genet 56, 5–7 (2011).
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The Pure Puerto Rican Paso Fino is an endangered breed of horse as recognized by The Livestock Conservancy and Equus Survival Trust.
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