ISSN 2074-9414 (Print),
ISSN 2313-1748 (Online)

Melanophilin Polymorphism of Exons 8 and 13 in Rabbits: Parent Breeds and Interbreed Cross Rodnik

Abstract
New breeds of fur-bearing animals are multi-breed crosses. Selection requires new methods for genetic control. This article describes a new early-maturing and highly-productive crossbreed of rabbits called Rodnik obtained at the Afanasyev Research Institute for Fur Farming and Rabbit Breeding, Russia. It was a result of multi-stage generations of crossings between such breeds as White Giant, Soviet Chinchilla, and Californian. However, in terms of polylocus genotypes of highly polymorphic elements, the Rodnik rabbits are closer to the White Giant breed. The article describes the changes in the genetic pool of this three-breed cross through a comparative analysis of the nucleotide sequences of exons 8 and 13 of the melanophilin (mlph) color gene. The research featured the rabbit breeds of White Giant, Soviet Chinchilla, and Californian, as well as their Rodnik cross. The analysis of the mlph gene in Oryctolagus cuniculus involved bioinformatics methods based on the nucleotide sequence of the reference genome UM_NZW_1.0 (GCF_009806435.1) from the GenBank database. The exon-intron structure was determined using the Splign software; the conditions and stages of PCR, including the design of primers for exonic regions, were developed using the online version of BLAST3. Exons 8 and 13 made it possible to identify the differences between chinchilla-colored rabbits and other breeds. In some color properties, the Rodnik cross was closer to the Californian parent breed. A single nucleotide polymorphism (SNP) was detected in two nucleotides in exon 8 and in one nucleotide in exon 13. The haplotypes of two SNPs in exon 8 and one SNP in exon 13 of the mlph gene were homozygous and coincided with the White Giant breed. The Soviet Chinchilla and Californian breeds carried a heterozygous haplotype for two SNPs in exon 8, as well as in the homozygote of another nucleotide (C–T) in exon 13 in the same positions. The identified SNPs did not correlate with color. However, the Rodnik cross was closer to the White Giant in terms of genotypes and polymorphisms of some highly polymorphic genomic elements. The mlph gene might become part of selection as it belongs to the Ras oncogene superfamily and the largest exophilin subfamily of Rab effector proteins that coordinate vesicular transport and some adipogenesis stages.
Keywords
Rabbits, breeding, population-genetic structure, melanophilin (mlph), SNP, fur color, intracellular organelles, crossbreed
REFERENCES
  1. Carneiro M, Afonso S, Geraldes A, Garreau H, Bolet G, et al. The genetic structure of domestic rabbits. Molecular Biology and Evolution. 2011;28(6):1801–1816. https://doi.org/10.1093/molbev/msr003
  2. Shchukina ES, Kosovsky GYu, Glazko VI, Kashapova IS, Glazko TT. Domestic rabbit Oryctolagus cuniculus var. domestica L. as a model in the study of domestication and biomedical researches (Review). The Agricultural Biology. 2020;55(4):643–658. (In Russ.) https://doi.org/10.15389/agrobiology.2020.4.643rus
  3. Dorożyńska K, Maj D. Rabbits – their domestication and molecular genetics of hair coat development and quality. Animal Genetics. 2021;52(1):10–20. https://doi.org/10.1111/age.13024
  4. Bennett DC, Lamoreux ML. The color loci of mice – A genetic century. Pigment Cell Research. 2003;16(4):333–344. https://doi.org/10.1034/j.1600-0749.2003.00067.x
  5. Alshanbari F, Castaneda C, Juras R, Hillhouse A, Mendoza MN, et al. Comparative FISH-mapping of MC1R, ASIP, and TYRP1 in New and Old World camelids and association analysis with coat color phenotypes in the dromedary (Camelus dromedarius). Frontiers in Genetics. 2019;10:340. https://doi.org/10.3389/fgene.2019.00340
  6. Jia X, Ding P, Chen S, Zhao S, Wang J, et al. Analysis of MC1R, MITF, TYR, TYRP1, and MLPH genes polymorphism in four rabbit breeds with different coat colors. Animals. 2021;11(1):81. https://doi.org/10.3390/ani11010081
  7. Neto MV, Hall MJ, Charneca J, Escrevente C, Seabra MC, et al. Photoprotective melanin is maintained within keratinocytes in storage lysosomes. Journal of Investigative Dermatology. 2025;145(5):1155–1165.e3. https://doi.org/10.1016/j.jid.2024.08.023
  8. Schiaffino MV. Signaling pathways in melanosome biogenesis and pathology. The International Journal of Biochemistry & Cell Biology. 2010;42(7):1094–1104. https://doi.org/10.1016/j.biocel.2010.03.023
  9. Morstein J, Bowcut V, Fernando M, Yang Y, Zhu L, et al. Targeting Ras-, Rho-, and Rab-family GTPases via a conserved cryptic pocket. Cell. 2024;187(22):6379–6392.e17. https://doi.org/10.1016/j.cell.2024.08.017
  10. Fukuda M, Kuroda TS, Mikoshiba K. Slac2-a/Melanophilin, the missing link between Rab27 and myosin Va: Implications of a tripartite protein complex for melanosome transport. Journal of Biological Chemistry. 2002;277(14):12432–12436. https://doi.org/10.1074/jbc.C200005200
  11. Kim D-H, Lee J, Ko J-K, Lee K. Melanophilin regulates dendritogenesis in melanocytes for feather pigmentation. Communications Biology. 2024;7:592. https://doi.org/10.1038/s42003-024-06284-5
  12. Martel JA, Michael D, Fejes-Tóth G, Náray-Fejes-Tóth A. Melanophilin, a novel aldosterone-induced gene in mouse cortical collecting duct cells. American Journal of Physiology-Renal Physiology. 2007;293(3):F904–F913. https://doi.org/10.1152/ajprenal.00365.2006
  13. Posbergh CJ, Staiger EA, Huson HJ. A stop-gain mutation within MLPH is responsible for the lilac dilution observed in jacob sheep. Genes. 2020;11(6):618. https://doi.org/10.3390/genes11060618
  14. Kosovsky GYu, Glazko VI, Abramov OI, Glazko TT. Melanophilin polymorphism in ferrets of different color. Doklady Biochemistry and Biophysics. 2023;513(Suppl 1):S12–S17. https://doi.org/10.1134/S1607672923700655
  15. Barral DC, Seabra MC. The melanosome as a model to study organelle motility in mammals. Pigment Cell Research. 2004;17(2):111–118. https://doi.org/10.1111/j.1600-0749.2004.00138.x
  16. Mercer JA, Seperack PK, Strobel MC, Copeland NG, Jenkins NA. Novel myosin heavy chain encoded by murine dilute coat colour locus. Nature. 1991;349:709–713. https://doi.org/10.1038/349709a0
  17. Wilson SM, Yip R, Swing DA, O'Sullivan TN, Zhang Y, et al. A mutation in Rab27a causes the vesicle transport defects observed in ashen mice. PNAS. 2000;97(14):7933–7938. https://doi.org/10.1073/pnas.140212797
  18. Matesic LE, Yip R, Reuss AE, Swing DA, O'Sullivan TN, et al. Mutations in Mlph, encoding a member of the Rab effector family, cause the melanosome transport defects observed in leaden mice. PNAS. 2001;98(18):10238–10243. https://doi.org/10.1073/pnas.181336698
  19. Lehner S, Gähle M, Dierks C, Stelter R, Gerber J, et al. Two-exon skipping within MLPH is associated with coat color dilution in rabbits. PLOS One. 2013;8(12):e84525. https://doi.org/10.1371/journal.pone.0084525
  20. Fontanesi L, Scotti E, Allain D, Dall'olio S. A frameshift mutation in the Melanophilin gene causes the dilute coat colour in rabbit (Oryctolagus cuniculus) breeds. Animal Genetics. 2014;45(2):248–255. https://doi.org/10.1111/age.12104
  21. Demars J, Iannuccelli N, Utzeri VJ, Auvinet G, Riquet J, et al. New insights into the Melanophilin (MLPH) gene affecting coat color dilution in rabbits. Genes. 2018;9(9):430. https://doi.org/10.3390/genes9090430
  22. Li J, Chen Y, Liu M, Chen Q, Zhou J, et al. Association of Melanophilin (MLPH) gene polymorphism with coat colour in Rex rabbits. World Rabbit Science. 2020;28(1):29–38. https://doi.org/10.4995/wrs.2020.12082
  23. Diribarne M, Mata X, Chantry-Darmon C, Vaiman A, Auvinet G, et al. A deletion in exon 9 of the LIPH gene is responsible for the Rex hair coat phenotype in rabbits (Oryctolagus cuniculus). PLOS One. 2011;6(4):e19281. https://doi.org/10.1371/journal.pone.0019281
  24. Shumilina AR. Dynamics of productive indicators of rabbits when creating the final three-breed cross. Krolikovodstvo i zverovodstvo. 2019;(6):9–15. (In Russ.) https://elibrary.ru/KODPJY
  25. Kosovsky GYu, Tinaev NI, Balakirev NA, Glazko VI. Rabbit breeding. Moscow: Moskovskij Dvor, 2023. 352 p. (In Russ.)
  26. Kolesnik ES, Glazko VI, Glazko TT, Kosovsky GYu. Intraspecific differentiation of the domestic rabbit (Oryctolagus cuniculus) using IRAP-PCR. Krolikovodstvo i zverovodstvo. 2022;(5):33–42. (In Russ.) https://doi.org/10.52178/00234885_2022_5_33
  27. Bueschbell B, Manga P, Schiedel AC. The many faces of G protein-coupled receptor 143, an atypical intracellular receptor. Frontiers in Molecular Biosciences. 2022;9:873777. https://doi.org/10.3389/fmolb.2022.873777
  28. Dai Y, Hu S, Bai S, Li J, Yang N, et al. CDK1 promotes the proliferation of melanocytes in Rex rabbits. Genes & Genomics. 2022;44:1191–1199. https://doi.org/10.1007/s13258-022-01283-4
  29. Wei M, Yang X, Yang X, Huang Y, Yuan Z, et al. MLPH regulates EMT in pancreatic adenocarcinoma through the PI3K-AKT signaling pathway. Journal of Cancer. 2024;15(17):5828–5838. https://doi.org/10.7150/jca.94573
  30. Tanwar J, Ahuja K, Sharma A, Sehgal P, Ranjan G, et al. Mitochondrial calcium uptake orchestrates vertebrate pigmentation via transcriptional regulation of keratin filaments. PLOS Biol. 2024;22(11):e3002895. https://doi.org/10.1371/journal.pbio.3002895
  31. Sharma N, Sharma A, Motiani RK. A novel gain of function mutation in TPC2 reiterates pH-pigmentation interplay: Emerging role of ionic homeostasis as a master pigmentation regulator. Cell Calcium. 2023;111:102705. https://doi.org/10.1016/j.ceca.2023.102705
  32. Kim M-Y, Kim Y-H, Park E-R, Shin Y, Kim GH, et al. MLPH is a novel adipogenic factor controlling redox homeostasis to inhibit lipid peroxidation in adipocytes. Biochemical and Biophysical Research Communications. 2024;734:150459. https://doi.org/10.1016/j.bbrc.2024.150459
  33. Wang H, Mizuno K, Takahashi N, Kobayashi E, Shirakawa J, et al. Melanophilin accelerates insulin granule fusion without predocking to the plasma membrane. Diabetes. 2020;69(12):2655–2666. https://doi.org/10.2337/db20-0069
  34. Chon NL, Tran S, Miller CS, Lin H, Knight JD. A conserved electrostatic membrane-binding surface in synaptotagmin-like proteins revealed using molecular phylogenetic analysis and homology modeling. Protein Science. 2024;33(1):e4850. https://doi.org/10.1002/pro.4850
  35. Babina M, Franke K, Bal G. How "neuronal" are human skin mast cells? International Journal of Molecular Sciences. 2022;23(18):10871. https://doi.org/10.3390/ijms231810871
  36. Macharadze DSh. Mast cells and tryptase. modern aspects. Medical Immunology (Russia). 2021;23(6):1271–1284. (In Russ.) https://doi.org/10.15789/1563-0625-MCA-2193
  37. Grigorev IP, Korzhevskii DE. Mast cells in the vertebrate brain: localization and functions. Zhurnal evolyutsionnoi biokhimii i fiziologii. 2021;57(1):17–32. (In Russ.) https://doi.org/10.31857/S0044452921010046
  38. Castaño-Jaramillo LM, Lugo-Reyes SO, Cruz Muñoz ME, Scheffler-Mendoza SC, Duran McKinster C, et al. Diagnostic and therapeutic caveats in Griscelli syndrome. Scandinavian Journal of Immunology. 2021;93(6):e13034. https://doi.org/10.1111/sji.13034
  39. Kim DH, Lee J, Suh Y, Chen PR, Lee K. Up-regulation of Melanophilin (MLPH) gene during avian adipogenesis and decreased fat pad weights with adipocyte hypotrophy in MLPH knockout quail. Poultry Science. 2024;104(2):104720. https://doi.org/10.1016/j.psj.2024.104720
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