Derived variants at six genes explain nearly half of size reduction in dog breeds

Selective breeding of dogs by humans has generated extraordinary diversity in body size. A number of multibreed analyses have been undertaken to identify the genetic basis of this diversity. We analyzed four loci discovered in a previous genome-wide association study that used 60,968 SNPs to identify size-associated genomic intervals, which were too large to assign causative roles to genes. First, we performed fine-mapping to define critical intervals that included the candidate genes GHR, HMGA2, SMAD2, and STC2, identifying five highly associated markers at the four loci. We hypothesize that three of the variants are likely to be causative. We then genotyped each marker, together with previously reported size-associated variants in the IGF1 and IGF1R genes, on a panel of 500 domestic dogs from 93 breeds, and identified the ancestral allele by genotyping the same markers on 30 wild canids. We observed that the derived alleles at all markers correlated with reduced body size, and smaller dogs are more likely to carry derived alleles at multiple markers. However, breeds are not generally fixed at all markers; multiple combinations of genotypes are found within most breeds. Finally, we show that 46%-52.5% of the variance in body size of dog breeds can be explained by seven markers in proximity to exceptional candidate genes. Among breeds with standard weights <41 kg (90 lb), the genotypes accounted for 64.3% of variance in weight. This work advances our understanding of mammalian growth by describing genetic contributions to canine size determination in non-giant dog breeds.

[1]  P. Clayton,et al.  A dominant-negative mutation of the growth hormone receptor causes familial short stature , 1997, Nature Genetics.

[2]  C. Heldin,et al.  The regulation of TGFβ signal transduction , 2009, Development.

[3]  M. Stephens,et al.  Accounting for Decay of Linkage Disequilibrium in Haplotype Inference and Missing-data Imputation , 2022 .

[4]  James A. Cuff,et al.  Genome sequence, comparative analysis and haplotype structure of the domestic dog , 2005, Nature.

[5]  V. Hwa,et al.  Evidence for a continuum of genetic, phenotypic, and biochemical abnormalities in children with growth hormone insensitivity. , 2011, Endocrine reviews.

[6]  Inês Barroso,et al.  Meta-Analysis of Genome-Wide Scans for Human Adult Stature Identifies Novel Loci and Associations with Measures of Skeletal Frame Size , 2009, PLoS genetics.

[7]  M. Hofreiter,et al.  Fossil dogs and wolves from Palaeolithic sites in Belgium, the Ukraine and Russia: osteometry, ancient DNA and stable isotopes , 2009 .

[8]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[9]  J. Lundeberg,et al.  Genetic Evidence for an East Asian Origin of Domestic Dogs , 2002, Science.

[10]  Bjarni V. Halldórsson,et al.  Many sequence variants affecting diversity of adult human height , 2008, Nature Genetics.

[11]  D. Haussler,et al.  Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomes. , 2005, Genome research.

[12]  K. Chada,et al.  Mini-mouse: phenotypic characterization of a transgenic insertional mutant allelic to pygmy. , 1994, Genetical research.

[13]  Manuel A. R. Ferreira,et al.  Assumption-Free Estimation of Heritability from Genome-Wide Identity-by-Descent Sharing between Full Siblings , 2006, PLoS genetics.

[14]  Natalie,et al.  Genetic Structure of the Purebred Domestic Dog , 2004 .

[15]  P. Green,et al.  Consed: a graphical tool for sequence finishing. , 1998, Genome research.

[16]  G. Crooks,et al.  WebLogo: a sequence logo generator. , 2004, Genome research.

[17]  J. Lundeberg,et al.  mtDNA Data Indicate a Single Origin for Dogs South of Yangtze River, Less Than 16,300 Years Ago, from Numerous Wolves , 2009, Molecular biology and evolution.

[18]  P. Rogalla,et al.  HMGI-C expression patterns in human tissues. Implications for the genesis of frequent mesenchymal tumors. , 1996, The American journal of pathology.

[19]  Hadley Wickham,et al.  ggplot2 - Elegant Graphics for Data Analysis (2nd Edition) , 2017 .

[20]  C. Hill,et al.  Tgf-beta superfamily signaling in embryonic development and homeostasis. , 2009, Developmental cell.

[21]  Xianjin Zhou,et al.  Mutation responsible for the mouse pygmy phenotype in the developmentally regulated factor HMGI-C , 1995, Nature.

[22]  Mark Daly,et al.  Haploview: analysis and visualization of LD and haplotype maps , 2005, Bioinform..

[23]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[24]  E. Ostrander,et al.  Morphometrics within dog breeds are highly reproducible and dispute Rensch’s rule , 2008, Mammalian Genome.

[25]  C. Gieger,et al.  Identification of ten loci associated with height highlights new biological pathways in human growth , 2008, Nature Genetics.

[26]  Pedro M. Valero-Mora,et al.  ggplot2: Elegant Graphics for Data Analysis , 2010 .

[27]  David M. Evans,et al.  Genome-wide association analysis identifies 20 loci that influence adult height , 2008, Nature Genetics.

[28]  Jennifer R. Harris,et al.  Combined Genome Scans for Body Stature in 6,602 European Twins: Evidence for Common Caucasian Loci , 2007, PLoS genetics.

[29]  Manuel A. R. Ferreira,et al.  PLINK: a tool set for whole-genome association and population-based linkage analyses. , 2007, American journal of human genetics.

[30]  B. Fogle The Encyclopedia of the Dog , 1995 .

[31]  Thomas J. Nicholas,et al.  Tracking footprints of artificial selection in the dog genome , 2010, Proceedings of the National Academy of Sciences.

[32]  E. Hardeman,et al.  The murine stanniocalcin 2 gene is a negative regulator of postnatal growth. , 2008, Endocrinology.

[33]  G. Novelli,et al.  Laron dwarfism and mutations of the growth hormone-receptor gene. , 1989, The New England journal of medicine.

[34]  Frederick R Adler,et al.  Interaction between the X chromosome and an autosome regulates size sexual dimorphism in Portuguese Water Dogs. , 2005, Genome research.

[35]  Christina Gloeckner,et al.  Modern Applied Statistics With S , 2003 .

[36]  P. Donnelly,et al.  A new statistical method for haplotype reconstruction from population data. , 2001, American journal of human genetics.

[37]  P Green,et al.  Base-calling of automated sequencer traces using phred. II. Error probabilities. , 1998, Genome research.

[38]  J. Wojcik,et al.  Journal of Clinical Endocrinology and Metabolism Printed in U.S.A. Copyright © 1998 by The Endocrine Society Four Contiguous Amino Acid Substitutions, Identified in Patients with Laron Syndrome, Differently Affect the Binding Affinity and Intracellular Tr , 2022 .

[39]  Frederick R Adler,et al.  Genetic basis for systems of skeletal quantitative traits: Principal component analysis of the canid skeleton , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[40]  V. Hwa,et al.  Defects in growth hormone receptor signaling , 2007, Trends in Endocrinology & Metabolism.

[41]  E. Ostrander,et al.  The insulin-like growth factor 1 receptor (IGF1R) contributes to reduced size in dogs , 2012, Mammalian Genome.

[42]  J. Wit,et al.  Genetic Disorders in the Growth Hormone – Insulin-Like Growth Factor-I Axis , 2006, Hormone Research in Paediatrics.

[43]  Lynn E Eberly,et al.  Body size and predatory performance in wolves: is bigger better? , 2009, The Journal of animal ecology.

[44]  C. Morton,et al.  HMGIC expression in human adult and fetal tissues and in uterine leiomyomata , 1999, Genes, chromosomes & cancer.

[45]  Shah Ebrahim,et al.  Common variants in the GDF5-UQCC region are associated with variation in human height , 2008, Nature Genetics.

[46]  K. Silventoinen DETERMINANTS OF VARIATION IN ADULT BODY HEIGHT , 2003, Journal of Biosocial Science.

[47]  Gary K. Chen,et al.  Identification, Replication, and Fine-Mapping of Loci Associated with Adult Height in Individuals of African Ancestry , 2011, PLoS genetics.

[48]  M. Savage,et al.  Intrauterine growth retardation and postnatal growth failure associated with deletion of the insulin-like growth factor I gene. , 1996, The New England journal of medicine.

[49]  Nicholas A. Johnson,et al.  Genome-wide association study of body height in African Americans: the Women's Health Initiative SNP Health Association Resource (SHARe). , 2012, Human molecular genetics.

[50]  Ayellet V. Segrè,et al.  Hundreds of variants clustered in genomic loci and biological pathways affect human height , 2010, Nature.

[51]  J. E. Rice,et al.  Multiple and ancient origins of the domestic dog. , 1997, Science.

[52]  P. Green,et al.  Base-calling of automated sequencer traces using phred. I. Accuracy assessment. , 1998, Genome research.

[53]  C. Bustamante,et al.  A Single IGF1 Allele Is a Major Determinant of Small Size in Dogs , 2007, Science.

[54]  B. Song,et al.  Smad signaling in skeletal development and regeneration. , 2009, Cytokine & growth factor reviews.

[55]  E. Ostrander,et al.  Single-Nucleotide-Polymorphism-Based Association Mapping of Dog Stereotypes , 2008, Genetics.

[56]  R. Wayne Molecular evolution of the dog family. , 1993, Trends in genetics : TIG.

[57]  C. Mammucari,et al.  Smad2 and 3 transcription factors control muscle mass in adulthood. , 2009, American journal of physiology. Cell physiology.

[58]  L. Andersson,et al.  Epistasis and the release of genetic variation during long-term selection , 2006, Nature Genetics.

[59]  J. van der Plicht,et al.  A 33,000-Year-Old Incipient Dog from the Altai Mountains of Siberia: Evidence of the Earliest Domestication Disrupted by the Last Glacial Maximum , 2011, PloS one.

[60]  K. Lindblad-Toh,et al.  Identification of Genomic Regions Associated with Phenotypic Variation between Dog Breeds using Selection Mapping , 2011, PLoS genetics.

[61]  D. Reich,et al.  Principal components analysis corrects for stratification in genome-wide association studies , 2006, Nature Genetics.

[62]  B. Wilcox,et al.  Atlas of Dog Breeds of the World , 1995 .

[63]  Richa Saxena,et al.  A common variant of HMGA2 is associated with adult and childhood height in the general population , 2007, Nature Genetics.

[64]  C. Deng,et al.  TGF-β and BMP Signaling in Osteoblast Differentiation and Bone Formation , 2012, International journal of biological sciences.

[65]  Mikhail V. Sablin,et al.  The Earliest Ice Age Dogs: Evidence from Eliseevichi 11 , 2002, Current Anthropology.

[66]  E. Ostrander,et al.  Canine Morphology: Hunting for Genes and Tracking Mutations , 2010, PLoS biology.

[67]  B. Han,et al.  Identification of 15 loci influencing height in a Korean population , 2010, Journal of Human Genetics.

[68]  G. F. Wagner,et al.  Human stanniocalcin-2 exhibits potent growth-suppressive properties in transgenic mice independently of growth hormone and IGFs. , 2005, American journal of physiology. Endocrinology and metabolism.

[69]  E. Kirkness,et al.  Breed relationships facilitate fine-mapping studies: a 7.8-kb deletion cosegregates with Collie eye anomaly across multiple dog breeds. , 2007, Genome research.

[70]  D. Reich,et al.  Population Structure and Eigenanalysis , 2006, PLoS genetics.

[71]  W. A. Bruette,et al.  The Complete Dog Book , 2008 .

[72]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[73]  Jeremiah D. Degenhardt,et al.  A Simple Genetic Architecture Underlies Morphological Variation in Dogs , 2010, PLoS biology.

[74]  Jeremiah D. Degenhardt,et al.  Genome-wide SNP and haplotype analyses reveal a rich history underlying dog domestication , 2010, Nature.

[75]  Elizabeth J. Robertson,et al.  Role of insulin-like growth factors in embryonic and postnatal growth , 1993, Cell.

[76]  D. Nickerson,et al.  PolyPhred: automating the detection and genotyping of single nucleotide substitutions using fluorescence-based resequencing. , 1997, Nucleic acids research.