Overview
Our lab seeks to identify genes that regulate bone density, size and shape. All of these skeletal phenotypes have been shown to be genetically regulated and to contribute to fracture susceptibility. To better understand the interaction between genes and environment in determining skeletal integrity, we use inbred strains of mice with skeletal differences as well as mice with spontaneous mutations that result in skeletal abnormalities. Our lab also studies the structure of the skull and face, and develops new spontaneous mutations in mice with craniofacial dysmorphologies. Our goal is to identify and characterize reliable genetic and physiological mouse models for human craniofacial disorders and to share these and other models for human skeletal diseases with the scientific public.
Research details
Mouse Models of Skeletal and Craniofacial Development and Morphology
Quantitative trait loci analyses of bone density and body composition in mice with growth hormone deficiency
Quantitative Trait Loci (QTL) analyses are powerful tool for the discovery of regulatory genes for complex traits such as body composition and skeletal phenotypes. Crosses between inbred strains of mice have provided data that are contributing to new genetic insights regarding bone and body morphology. Further, QTL analyses can be used to partition phenotypes into their regulatory pathways by using mice with altered genomes. We use the little (Ghrhrlit) mutation, a model of growth hormone and IGF-1 deficiency, which arose spontaneously on the C57BL/6J (B6) low-bone-density inbred strain background, coupled with a new congenic strain we made by introgressing the little mutation onto the high-bone-density C3H/HeJ (C3H) inbred strain background. Homozygous mice on both backgrounds, (B6-lit/lit and C3.B6-lit/lit), have undetectable circulating growth hormone (GH), low serum insulin-like growth factor 1 (IGF1), and reduced bone mineral density (BMD), femur length, and body mass compared to normal littermates. However, although C3.B6-lit/lit mice are of the same body weight and femur length as B6-lit/lit mice, C3.B6-lit/lit mice have higher BMD and female C3.B6-lit/lit mice have lower percent body fat than B6-lit/lit females, while males of the two strains do not differ in body fat mass. To explore the genetic regulation of these differences, we crossed homozygous mutant mice of the two little strains, B6-lit/lit and C3.B6-lit/lit, to produce 1,008 male and 1,062 female F2 mice, all GH/IGF-1 deficient, but segregating alleles from B6 and C3H. QTL analyses of genome-wide data from 120 polymorphic markers (nearly 211,761 genotypes) revealed 14 regulatory loci for total body BMD, 14 for femoral BMD, 15 for periosteal circumference, 14 for femur length, 8 for L2-L4 lumbar vertebral BMD, 14 for body weight, 12 for percent body fat, and 11 for serum IGF-1 with highly significant LOD scores. All nineteen chromosomes had QTL for multiple phenotypes.
Further, we have identified interactive pairs of QTL in which a particular QTL is not detected until it is paired with another QTL. For example, the QTL on Chromosome (Chr) 2 at 26cM for whole body BMD was not detected in the main effect analysis; however the interactive pair 2:18 (Chr 18 at 42cM) has a significant LOD of 5.09. Again for the whole body BMD phenotype, the QTL on Chr 15 must be paired with either the QTL on Chr 5 or 7 to have a significant effect. In contrast, the contributing QTL to the interactive pairs Chr 9:11 and 12:16 each has a significant individual effect, as well as a significant interactive effect. Similar examples can be found for other phenotypes. The importance of these interacting QTL is threefold: the proteins encoded by the genes within the QTL may interact physically, e.g., ligand and receptor; they may represent a biological pathway; or they may be part of a redundant physiological system. These possibilities can only be evaluated once the genes within the QTL are identified; this work is ongoing in my laboratory.
Mouse models of craniofacial disorders
In addition to the axial and appendicular skeleton, my laboratory studies craniofacial morphology. Our goal is to identify and characterize reliable genetic and physiological mouse models for human craniofacial dysmorphologies and to share these models with the scientific public. To date, we have identified more than one hundred deviants with heritable craniofacial abnormalities including: abnormal ear pinnae shape, size, and placement; shortened snouts; shortened, domed, and asymmetrical skulls; abnormal dentition; abnormal eye morphology; and deafness. We perform comprehensive genetic studies and preliminary characterization studies that include clinical examination, basic histology, and evaluation of bone-density and skeletal anomalies, dentition, vision, and hearing. Recently, we have identified the gene responsible for three new mutants: one with extra teeth, one with glaucoma, and one with a short lower jaw, small ears, and a sparse coat. Manuscripts describing these new mouse models and their importance to human health are in preparation. In addition, the chromosomal locations of several other new mutants have been identified. We provide both uncharacterized and completed models to the scientific community via our web site www.jax.org/cranio/ and distribute live mice upon request. We have a dedicated email address, faces@jax.org, for direct communication with interested investigators.
Mouse models of fracture susceptibility
The skeleton serves ambulatory, as well as metabolic and protective, functions. Because bone is important for mobility and muscle attachment, the ultimate test of skeletal integrity is its resistance to fracture. Although fracture often is associated with osteoporotic patients, stress fracture from constant overuse and acute fracture from trauma can occur in any population. In the human clinic, the gold standard used to predict fracture risk is bone mineral density (BMD) as measured by dual energy X-ray absorptiometry. Yet other characteristics of bone architecture including size, shape, and quality are important contributors to bone strength and should be considered when predicting fracture risk. In addition, it has been shown that these morphological phenotypes are often associated with differences in muscle mass. Coupled with this expanded interest in phenotyping of bone and muscle is an intense search for genetic predisposition for fracture based on these newly identified indices of bone strength. The interpretation of these studies has been complicated by the heterogeneity of the human population and by environmental differences. Taken together, however, the studies demonstrate that genetic regulation of bone strength is complex and that animal models are critical to progress in this field.
To determine if BMD, muscle mass, or bone morphological phenotypes are good predictors of skeletal strength in mice, we collected data on skeletal geometry, muscle mass, BMD, and bone strength from mice of nine genetically diverse inbred strains. We used three measures of bone strength: peak load (the maximum amount of stress that the femur can withstand before breaking); stiffness (the amount of stress the bone can withstand and still return to its original shape); and energy to break (a measure of the amount of energy needed to cause a fracture, often termed the toughness of bone).
We found that BMD at the mid-shaft of the femur was a significant predictor of peak load and stiffness, but was a poor predictor of energy to break. In contrast, measures of femoral geometry-femur length and dimension-were significant predictors of peak load, stiffness, and energy to break. Similarly, both the area and weight of the quadricep muscle were good predictors of all three measures of bone strength in both male and female mice, although the weight of the isolated quadricep muscle was a stronger predictor of energy to break in females than in males, and a better predictor of peak load in males than in females. These data support the hypothesis that skeletal geometry and muscle mass are important determinants of bone strength and fracture risk. In addition, we have shown that there are sex differences for some predictors of bone strength. Finally, our data demonstrate that inbred strains of mice can be used as models to discover the genetic regulation of bone geometry and strength.
Mouse models of heritable skeletal diseases
With the Mouse Mutant Resource, we continue to develop mouse models for skeletal diseases that result from spontaneous mutations in single genes. We are currently characterizing and mapping several new mutations that result in appendicular and axial skeletal malformations, craniofacial abnormalities, and both loss and abnormal accretion of bone mineral. These mutants will become important models for studying bone physiology and morphology.
Lab staff
Principal Investigator: Leah Rae Donahue, Ph.D.Co-Principal Investigators: Wesley Beamer, Ph.D., Muriel T. Davisson, Ph.D., Clifford J. Rosen, M.D.
Research Assistant II: Michelle M. Curtain, B.A.
Biomedical Technologist II: Coleen Marden
Biomedical Technologist I: Julie Hurd
Publication listings
Beamer WG, Donahue LR, Rosen CJ. 2002. Genetics and bone using the mouse to understand man. J Musculoskel Neuron Interact 2:225-231.
Bouxsein ML, Rosen CJ, Turner CH, Ackert CL, Shultz KL, Donahue LR, Churchill G, Adamo M, Powell DR, Turner RT, Muller R, Beamer WG. 2002. Generation of a new congenic mouse strain to test the relationships among serum insulin-like growth factor I, bone mineral density, and skeletal morphology in vivo. J Bone Miner Res 17:570-579.
Cotman SL, Vrbanac V, Lebel L-A, Lee RL, Johnson KA, Donahue LR, Teed AM, Antonellis K, Bronson RT, Lerner TJ, MacDonald ME. 2002. Cln3deltaex7/8 knock-in mice with the common JNCL mutation exhibit progressive neurologic disease that begins before birth. Hum Molec Genet 11:2709-2721.
Gao H, Boustany R-MN, Espinola JA, Cotman SL, Srinidhi L, Antonellis KA, Gillis T, Qin X, Liu S, Donahue LR, Bronson RT, Faust JR, Stout D, Haines JL, Lerner TJ, MacDonald ME. 2002. Mutations in a novel CLN6-encoded transmembrane protein cause variant neuronal ceroid lipofuscinosis in man and mouse. Am J Hum Genet 70:324-335.
Gu WK, Li XM, Edderkaoui B, Strong DD, Lau K-HW, Beamer WG, Donahue LR, Mohan S, Baylink DJ. 2002. Construction of a BAC contig for a 3cM biologically significant region of mouse chromosome 1. Genetica 114:1-9.
Judex S, Donahue LR, Rubin C. 2002. Genetic predisposition to low bone mass is paralleled by an enhanced sensitivity to signals anabolic to the skeleton. Faseb J 16:1280-1282.
Sheng MH, Baylink DJ, Beamer WG, Donahue LR, Lau KH, Wergedal JE. 2002. Regulation of bone volume is different in the metaphyses of the femur and vertebra of C3H/HeJ and C57BL/6J mice. Bone 30(3):486-491.
Bauschatz JD, Curtain MM, Davisson MT, Lane PW, Donahue LR. 2003. In: Collaboration: The Jackson Laboratory Craniofacial Resource, Special Issue: Honoring Dr. Sandy C. Marks, Jr., 1937-2002. Crit Rev Eukaryot Gene Expr 13(2-4):107-108.
Donahue LR, Chang B, Subburaman M, Miyakoshi N, Wergedal JE, Baylink DJ, Hawes NL, Rosen CJ, Ward-Bailey P, Zheng QY, Bronson RT, Johnson KR, Davisson MT. 2003. A missense mutation in the mouse Col2al gene causes spondyloepiphyseal dysplasia hearing loss, and retinoschisis. J Bone Miner Res 18(9):1612-1621.
Koller D, Schriefer J, Sun O, Shultz KL, Donahue LR, Rosen CJ, Foroud T, Beamer WG, Turner CH. 2003. Genetic effects for femoral biomechanics, structure, and density in C57BL/6J and C3H/HeJ inbred mouse strains. J Bone Miner Res 18(10):1758-1765.
Mohan S, Richman C, Guo R, Amaar Y, Donahue LR, Wergedal J, Baylink DJ. 2003. Insulin-like growth factor regulates peak bone mineral density in mice by both growth hormone-dependent and - independent mechanisms. Endocrinology 144(3):929-936.
Shultz KL, Donahue LR, Bouxsein ML, Baylink DJ, Rossen CJ, Beamer WG. 2003. Congenic strains of mice for verification and genetic decomposition of quantitative loci for femoral bone mineral density. J Bone Miner Res 18(2):175-185.
Turner CH, Sun O, Schriefer J, Pitner N, Price R, Bouxsein ML, Rosen CJ, Donahue LR, Shultz KL, Beamer WG. 2003. Congenic mice reveal sex-specific genetic regulation of femoral structure and strength. Calcif Tissue Int 73(3):297-303.
Bouxsein ML, Uchiyama T, Rosen CJ, Shultz KL, Donahue LR, Turner CH, Sen S, Churchill GA, Muller R, Beamer WG. 2004. Mapping quantitative trait Loci for vertebral trabecular bone volume fraction and microarchitecture in mice. J Bone Miner Res 19(4):587-599.
Ho M, Post CM, Donahue LR, Lidov HGW, Bronson RT, Goolsby H, Watkins SC, Cox GA, Brown RH Jr. 2004. Disruption of muscle membrane and phenotype divergence in two novel mouse models of dysferlin deficiency. Hum Mol Genet 13(18):1999-2010.
Judex S, Garman R, Squire M, Busa B, Donahue LR, Rubin C. 2004. Genetically linked site-specificity of disuse osteoporosis. J Bone Miner Res 19(4):607-613.
Judex S, Garman R, Squire M, Donahue LR, Rubin C. 2004. Genetically based influences on the site-specific regulation of trabecular and cortical bone morphology. J Bone Miner Res 19(4):600-606.
Lorenz-Depiereux B, Guido VE, Johnson KR, Zheng QY, Gagnon LH, Bauschatz JD, Davisson MT, Washburn LL, Donahue LR, Strom TM, Eicher EM. 2004. New intragenic deletions in the Phex gene clarify X-linked hypophosphatemia-related abnormalities in mice. Mamm Genome 15(3):151-161.
Rosen CJ, Ackert-Bicknell CL, Adamo ML, Shultz KL, Rubin J, Donahue LR, Horton LG, Horowitz MC. 2004. Congenic mice with low serum IGF-1 have increased body fat, reduced bone mineral density, and an altered osteoblast differentiation program. Bone 35:1046-1058.
Squire M, Donahue LR, Rubin CT, Judex S. 2004. Genetic variations that regulate bone morphology in the male skeleton do not influence its susceptibility to mechanical unloading. Bone 35(6):1353-1360.
Beamer WG, Shultz KL, Donahue LR, Churchill GA, Sen S, Wergedal JR, Baylink DJ, Rosen CJ. 2005. Quantitative trait loci for femoral and lumbar vertebral bonem mineral density in C57BL/6J and C3H/HeJ inbred strains of mice. J Bone Miner Res 20(9):1701-1712.
Bouxsein ML, Myers KS, Shultz KL, Donahue LR, Rosen CJ, Beamer WG. 2005. Ovariectomy-induced bone loss varies among inbred strains of mice. J Bone Miner Res 20(7):1085-1092.
Jiao Y, Yan J, Zhao Y, Donahue LR, Beamer WG, Li X, Roe BA, LeDoux MS, Gu W. 2005. Carbonic anhydrase-related protein VIII deficiency is associated with a distinctive lifelong gait disorder in waddles mice. Genetics 171:1239-1246.
Judex S, Zhong N, Squire ME, Donahue LR, Hadjiargyrou M, Rubin CT. 2005. Mechanical modulation of molecular signals which regulate anabolic and catabolic activity in bone tissue. J Cell Biochem 1(94):982-984.
Mohan S, Kapoor A, Singgih A, Zhang Z, Taylor T, Yu H, Chadwick RB, Chung Y-S, Donahue LR, Rosen C, Crawford GC, Wergedal J & Baylink DJ. 2005. Spontaneous fractures in the mouse mutant sfx are caused by deletion of the gulonolactone oxidase gene, causing vitamin C deficiency. J Bone Miner Res 20(9):1597-1610.
Zhong N, Garman R, Squire M, Donahue LR, Rubin C, Hadjiargyrou M, Judex S. 2005. Gene expression patterns in bone after 4 days of hind-limb unloading in two inbred strains of mice. Aviat Space Environ Med 76(6);530-535.
Adamo ML, Ma X, Ackert-Bicknell CL, Donahue LR, Beamer WG, Rosen CJ. 2006. Genetic increase in serum insulin-like growth factor-l(lGF-l) in C3H/HeJ compared with C67BL/6J mice is associated with increased transcription from the IGF-I exon 2 promoter. Endocrinology 147:2944-2955.
Akeson EC, Donahue LR, Beamer WG, Shultz KL, Ackert-Bicknell C, Rosen CJ, Corrigan J, Davisson MT. 2006. Chromosomal inversion discovered in C3H/HeJ mice. Genomics 87:311-313.
Delahunty KM, Shultz KL, Gronowicz GA, Koczon-Jaremko B, Adamo ML, Horton LG, Lorenzo J, Donahue LR, Ackert-Bicknell C, Kream BE, Beamer WG, Rosen CJ. 2006. Congenic mice provide in vivo evidence for a genetic locus that modulates serum insulin-like growth factor-land bone acquisition. Endocrinology 147(8):3915-3923.
Gilbert SL, Zhang L, Forster ML, Iwase T, Soliven Donahue LR, Sweet HO, Bronson RT, Davisson MT, Wollmann RL, Lahn BT. 2006. Trak1 mutation disrupts GABA(A) receptor homeostasis in hypertonic mice. Nat Genet 38(2):245-250.
Ishimori N, Li R, Walsh KA, Korstanje R, Rollins JA, Petkov P, Pletcher MT, Wiltshire T, Donahue LR, Rosen CJ, Beamer WG, Churchill GA, Paigen B. 2006. Quantitative trait loci that determine BMD in C57BL/6J and 129S1/SvlmJ inbred mice. J Bone Miner Res 21(1):105-112.
Xie L, Jacobson JM, Choi ES, Busa B, Donahue LR, Miller LM, Rubin CT, Judex S. 2006. Low-level mechanical vibrations can influence bone resorption and bone formation in the growing skeleton. Bone 39(5):1059-1066.
Beamer WG, Shultz KL, Ackert-Bicknell CL, Horton LG, Delahunty KM, Coombs III HF, Donahue LR, Canalis E, Rosen CJ. 2007. Genetic dissection of mouse distal chromosome 1 reveals three linked BMD QTLs with sex-dependent regulation of bone phenotypes. J Bone Miner Res 22(8):1187-1196.
Garman R, Gaudette G, Donahue LR, Rubin C, Judex S. 2007. Low-level accelerations applied in the absence of weight bearing can enhance trabecular bone formation. J Orthop Res 25:732-740.
Jiao Y, Yan J, Jiao F, Yang H, Donahue LR, Li X, Roe BA, Stuart J, Gu W. 2007. A single nucleotide mutation in Nppc is associated with long bone abnormality in lbab mice. BMC Genetics 8:16.
Johnson KR, Marden CC, Ward-Bailey P, Gagnon LH, Bronson RT, Donahue LR. 2007. Congenital hypothyroidism, dwarfism, and hearing impairment caused by a missense mutation in mouse dual oxidase 2 gene, Duox2. Mol Endocrinol 21(7):1593-1602.
Judex S, Rubin C, Gaudette G, Garman R, Donahue LR. 2007. Low-level accelerations applied in the absence of weight bearing can enhance trabecular bone formation. J Orthop Res 25(6):732-740.
Kohler T, Stauber M, Donahue LR, Muller R. 2007. Automated compartmental analysis for high-throughput skeletal phenotyping in femora of genetic mouse models. Bone 41:659-667.
Schneider P, Stauber M, Voide R, Stampanoni M, Donahue LR, Muller R. 2007. Ultrastructural properties in cortical bone vary greatly in two inbred strains of mice as assessed by synchrotron light based micro- and nano-CT. J Bone Miner Res 22(10):1557.
Yan J, Jiao Y, Jiao F, Stuart J, Donahue LR, Beamer WG, Li X, Roe BA, LeDoux MS, Gu W. 2007. Effects of carbonic anhydrase VIII deficiency on cerebellar gene expression profiles in the wdl mouse. Neurosci Lett 413:196-201.
Gregorova S, Divina P, Storchova R, Trachtulec Z, Fotopulosova V, Svenson KL, Donahue LR, Paigen B, Forejt J. 2008. Mouse consomic strains: Exploiting genetic divergence between Mus m. musculus and Mus m. domesticus subspecies. Genome Res 18(3):509-515.
Li R, Svenson KL, Donahue LR, Peters LL, Churchill GA. 2008. The relationships of dietary fat, body composition and bone mineral density in inbred mouse strain panels. Physiol Genomics 33(1):26-32.
