Comparison of horse chromosome 3 with donkey and human chromosomes by cross-species painting and heterologous FISH mapping.
Abstract: The melanocortin 1 receptor (MC1R), mast/stem cell growth factor receptor (KIT), and platelet-derived growth factor receptor alpha (PDGFRA) are loci that all belong to equine linkage group 2 (LG2). Of these, KIT was fluorescent in situ hybridization (FISH) mapped to ECA3q21 with equine cDNA and heterologous porcine BAC probes, while MC1R was localized to ECA3p12 and PDGFRA to ECA3q21 with heterologous porcine BAC probes. A three-step comparison between ECA3 and donkey chromosomes was carried out. First, microdissected ECA3 painting probe was used on donkey chromosomes, which showed disruption of the equine synteny. Next, human (HSA) Chromosomes (Chrs) 16q and 4 specific paints, known to be homologous to ECA3p and 3q, respectively, were applied to detect homologous chromosomal segment(s) in donkey. Finally, four genes (MC1R, ALB, PDGFRA, KIT) and two equine microsatellite markers (SGCV18 and SGCV33) located on ECA3 were FISH mapped to donkey chromosomes. The findings refined the cross species painting homology results and added six new markers to the nascent donkey gene map. The hypothesis that Tobiano coat color in horses may be associated with a chromosomal inversion involving genes within LG2 was tested by G-banding-based cytogenetic analysis and ordering of four loci-KIT, PDGFRA, albumin (ALB), and MC1R-in Tobiano and non-tobiano (homozygous as well as heterozygous) horses. However, no difference either in banding patterns or location/relative order of the genes was observed in the three classes. The study highlights successful FISH mapping of BAC probes across evolutionarily diverged species, viz., pig and horse/donkey, and represents the first use of large-sized individual clones across distantly related farm animals.
Publication Date: 1999-03-02 PubMed ID: 10051324DOI: 10.1007/s003359900986Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research focuses on the genetic comparisons and mapping of horse chromosome 3 with that of donkeys and humans. It finds that large-sized individual clones, or segments of DNA, can be successfully employed across distantly related farm animals like horses and pigs.
Chromosome Mapping and FISH technique
- The researchers analyzed specific genetic locations (KIT, MC1R, and PDGFRA) on equine linkage group 2 (LG2) using a technique known as fluorescent in situ hybridization (FISH). FISH works by using probes that “light up” under a fluorescent microscope, allowing the visualization of these genes within the cells.
- KIT, which plays a part in cell signalling and organ development, was mapped to ECA3q21. Simultaneously, MC1R – involved in the pigmentation and immune response – was localized to ECA3p12 and PDGFRA responsible for cell growth, to ECA3q21. This was accomplished using porcine (pig) BAC probes, which help identify the position of these genes on their corresponding chromosomes.
Comparisons and Findings
- The team undertook a trifold comparison between horse chromosome 3 and donkey chromosomes using a microdissected ECA3 painting probe, making evident a noticeable disruption of the equine synteny, or the linear sequence of genes.
- Next, by using specific paint markers for human chromosomes 16q and 4, homologous segments in the donkey chromosome were detected. These painted segments are known to be similar to corresponding parts of horse chromosome 3.
- Four particular genes (MC1R, ALB, PDGFRA, KIT) and two horse microsatellite markers were then successfully mapped onto the donkey chromosomes. This furthered the understanding of relatedness within species and added six new markers to the emerging donkey gene map.
The Role of Chromosomal Inversion
- The researchers also dived into the hypothesis linking the Tobiano coat color in horses to a chromosomal inversion involving genes within LG2, basically suggesting that flipping of the gene order could influence coat color. They compared the banding patterns and relative gene order of four loci (KIT, PDGFRA, ALB, and MC1R) in Tobiano and non-Tobiano horses.
- However, they found no difference in either the banding patterns or the order/location of genes across these horse types, debunking the initial hypothesis.
Significance of the Study
- This study demonstrates the successful application of FISH mapping (especially using large-sized individual clones) across distantly related species – in this case, the pig and the horse/donkey. It signifies a step forward in genetic research and understands the relatedness and differences between different species’ genes.
Cite This Article
APA
Raudsepp T, Kijas J, Godard S, Guérin G, Andersson L, Chowdhary BP.
(1999).
Comparison of horse chromosome 3 with donkey and human chromosomes by cross-species painting and heterologous FISH mapping.
Mamm Genome, 10(3), 277-282.
https://doi.org/10.1007/s003359900986 Publication
Researcher Affiliations
- Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Box 7023, S-750 07 Uppsala, Sweden.
MeSH Terms
- Animals
- Base Sequence
- Chromosomes
- Chromosomes, Human
- DNA Primers
- DNA Probes
- DNA, Complementary
- Equidae / genetics
- Horses / genetics
- Humans
- In Situ Hybridization, Fluorescence / methods
- Species Specificity
Citations
This article has been cited 12 times.- Finno CJ, Bannasch DL. Applied equine genetics.. Equine Vet J 2014 Sep;46(5):538-44.
- De Lorenzi L, Genualdo V, Perucatti A, Pia Di Meo G, Molteni L, Iannuzzi L, Parma P. Chromosomal assignment of R-spondin genes in the donkey (Equus asinus, 2n = 62).. J Appl Genet 2010;51(3):319-21.
- Trifonov VA, Stanyon R, Nesterenko AI, Fu B, Perelman PL, O'Brien PC, Stone G, Rubtsova NV, Houck ML, Robinson TJ, Ferguson-Smith MA, Dobigny G, Graphodatsky AS, Yang F. Multidirectional cross-species painting illuminates the history of karyotypic evolution in Perissodactyla.. Chromosome Res 2008;16(1):89-107.
- Chowdhary BP, Raudsepp T. The horse genome derby: racing from map to whole genome sequence.. Chromosome Res 2008;16(1):109-27.
- Raudsepp T, Santani A, Wallner B, Kata SR, Ren C, Zhang HB, Womack JE, Skow LC, Chowdhary BP. A detailed physical map of the horse Y chromosome.. Proc Natl Acad Sci U S A 2004 Jun 22;101(25):9321-6.
- Yang F, Fu B, O'Brien PC, Nie W, Ryder OA, Ferguson-Smith MA. Refined genome-wide comparative map of the domestic horse, donkey and human based on cross-species chromosome painting: insight into the occasional fertility of mules.. Chromosome Res 2004;12(1):65-76.
- Chowdhary BP, Raudsepp T, Kata SR, Goh G, Millon LV, Allan V, Piumi F, Guérin G, Swinburne J, Binns M, Lear TL, Mickelson J, Murray J, Antczak DF, Womack JE, Skow LC. The first-generation whole-genome radiation hybrid map in the horse identifies conserved segments in human and mouse genomes.. Genome Res 2003 Apr;13(4):742-51.
- Santani A, Raudsepp T, Chowdhary BP. Interstitial telomeric sites and NORs in Hartmann's zebra (Equus zebra hartmannae) chromosomes.. Chromosome Res 2002;10(7):527-34.
- Raudsepp T, Chowdhary BP. Correspondence of human chromosomes 9, 12, 15, 16, 19 and 20 with donkey chromosomes refines homology between horse and donkey karyotypes.. Chromosome Res 2001;9(8):623-9.
- Raudsepp T, Christensen K, Chowdhar BP. Cytogenetics of donkey chromosomes: nomenclature proposal based on GTG-banded chromosomes and depiction of NORs and telomeric sites.. Chromosome Res 2000;8(8):659-70.
- Metallinos D, Rine J. Exclusion of EDNRB and KIT as the basis for white spotting in Border Collies.. Genome Biol 2000;1(2):RESEARCH0004.
- Raudsepp T, Chowdhary BP. Construction of chromosome-specific paints for meta- and submetacentric autosomes and the sex chromosomes in the horse and their use to detect homologous chromosomal segments in the donkey.. Chromosome Res 1999;7(2):103-14.
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