Dorsoventral Patterning of the Mouse Coat by Tbx15

Many members of the animal kingdom display coat or skin color differences along their dorsoventral axis. To determine the mechanisms that control regional differences in pigmentation, we have studied how a classical mouse mutation, droopy ear (deH), affects dorsoventral skin characteristics, especially those under control of the Agouti gene. Mice carrying the Agouti allele black-and-tan (at) normally have a sharp boundary between dorsal black hair and yellow ventral hair; the deH mutation raises the pigmentation boundary, producing an apparent dorsal-to-ventral transformation. We identify a 216 kb deletion in deH that removes all but the first exon of the Tbx15 gene, whose embryonic expression in developing mesenchyme correlates with pigmentary and skeletal malformations observed in deH/deH animals. Construction of a targeted allele of Tbx15 confirmed that the deH phenotype was caused by Tbx15 loss of function. Early embryonic expression of Tbx15 in dorsal mesenchyme is complementary to Agouti expression in ventral mesenchyme; in the absence of Tbx15, expression of Agouti in both embryos and postnatal animals is displaced dorsally. Transplantation experiments demonstrate that positional identity of the skin with regard to dorsoventral pigmentation differences is acquired by E12.5, which is shortly after early embryonic expression of Tbx15. Fate-mapping studies show that the dorsoventral pigmentation boundary is not in register with a previously identified dermal cell lineage boundary, but rather with the limb dorsoventral boundary. Embryonic expression of Tbx15 in dorsolateral mesenchyme provides an instructional cue required to establish the future positional identity of dorsal dermis. These findings represent a novel role for T-box gene action in embryonic development, identify a previously unappreciated aspect of dorsoventral patterning that is widely represented in furred mammals, and provide insight into the mechanisms that underlie region-specific differences in body morphology.

[1]  Y. Lan,et al.  Isolation and developmental expression analysis of Tbx22, the mouse homolog of the human X‐linked cleft palate gene , 2002, Developmental dynamics : an official publication of the American Association of Anatomists.

[2]  M. Kuehn,et al.  HoxB6‐Cre transgenic mice express Cre recombinase in extra‐embryonic mesoderm, in lateral plate and limb mesoderm and at the midbrain/hindbrain junction , 2000, Genesis.

[3]  M. Evans,et al.  A combined analysis of genomic and primary protein structure defines the phylogenetic relationship of new members if the T-box family. , 1998, Genomics.

[4]  L. Silver,et al.  Cloning, mapping, and expression analysis of TBX15, a new member of the T-Box gene family. , 1998, Genomics.

[5]  R. Woychik,et al.  Molecular characterization of the mouse agouti locus , 1992, Cell.

[6]  H. Moriya,et al.  Inductive signals from the somatopleure mediated by bone morphogenetic proteins are essential for the formation of the sternal component of avian ribs. , 2001, Developmental biology.

[7]  O. Pourquié,et al.  Mediolateral somitic origin of ribs and dermis determined by quail-chick chimeras. , 2000, Development.

[8]  P. Mauger Rôle du mésoderme somitique dans le développement du plumage dorsal chez l'embryon de Poulet: I. Origine, capacités de régulation et détermination du mésoderme plumigène , 1972 .

[9]  Wolfgang Wurst,et al.  The mouse Engrailed-1 gene and ventral limb patterning , 1996, Nature.

[10]  B. Herrmann,et al.  Detection of messenger RNA by in situ hybridization to postimplantation embryo whole mounts. , 1993, Methods in enzymology.

[11]  C. Logan,et al.  The role of Engrailed in establishing the dorsoventral axis of the chick limb. , 1997, Development.

[12]  M. Tidball,et al.  The Coat Colors of Mice. A Model for Mammalian Gene Action and Interaction , 1980, The Yale Journal of Biology and Medicine.

[13]  M. Cohn,et al.  Fibroblast growth factors induce additional limb development from the flank of chick embryos , 1995, Cell.

[14]  N. L. Le Douarin,et al.  The dorsoventral polarity of the presumptive limb is determined by signals produced by the somites and by the lateral somatopleure. , 1997, Development.

[15]  A. Kispert,et al.  Cloning and expression analysis of the mouse T-box gene Tbx18 , 2001, Mechanisms of Development.

[16]  P. F. Forsthoefel,et al.  The origin and development of alopecia in mice homozygous for strong's luxoid gene , 1966, Journal of morphology.

[17]  Ralf J. Sommer,et al.  The evolution of signalling pathways in animal development , 2003, Nature Reviews Genetics.

[18]  M. Davisson,et al.  Molecular analysis of reverse mutations from nonagouti (a) to black-and-tan (a(t)) and white-bellied agouti (Aw) reveals alternative forms of agouti transcripts. , 1994, Genes & development.

[19]  A. Rump,et al.  Expression of mouse Tbx22 supports its role in palatogenesis and glossogenesis , 2003, Developmental dynamics : an official publication of the American Association of Anatomists.

[20]  Andrew P. McMahon,et al.  Dorsalizing signal Wnt-7a required for normal polarity of D–V and A–P axes of mouse limb , 1995, Nature.

[21]  A. Joyner,et al.  Two lineage boundaries coordinate vertebrate apical ectodermal ridge formation. , 2000, Genes & development.

[22]  A. Joyner,et al.  Analysis of the genetic pathway leading to formation of ectopic apical ectodermal ridges in mouse Engrailed-1 mutant limbs. , 1998, Development.

[23]  Philippe Soriano Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[24]  M. Patton,et al.  Craniofacial expression of human and murine TBX22 correlates with the cleft palate and ankyloglossia phenotype observed in CPX patients. , 2002, Human molecular genetics.

[25]  G. Barsh,et al.  Expression and transgenic studies of the mouse agouti gene provide insight into the mechanisms by which mammalian coat color patterns are generated. , 1995, Development.

[26]  C. Tickle,et al.  Engrailed-1 misexpression in chick embryos prevents apical ridge formation but preserves segregation of dorsal and ventral ectodermal compartments. , 2000, Developmental biology.

[27]  V. Papaioannou T-box genes in development: from hydra to humans. , 2001, International review of cytology.

[28]  J. Nordlund The pigmentary system : physiology and pathophysiology , 1998 .

[29]  D. Kaufman Adaptive coloration in Peromyscus polionotus experimental selection by owls , 1974 .

[30]  L. Shaffer,et al.  Haploinsufficiency of ALX4 as a potential cause of parietal foramina in the 11p11.2 contiguous gene-deletion syndrome. , 2000, American journal of human genetics.

[31]  J. Clarke,et al.  Dorso-ventral ectodermal compartments and origin of apical ectodermal ridge in developing chick limb. , 1997, Development.

[32]  J. Smith,et al.  T‐targets: Clues to understanding the functions of T‐box proteins , 2001, Development, growth & differentiation.

[33]  Thomas M. Jessell,et al.  Induction of the LIM homeobox gene Lmx1 by WNT6a establishes dorsoventral pattern in the vertebrate limb , 1995, Cell.

[34]  P. Goodfellow,et al.  The T-box transcription factor gene TBX22 is mutated in X-linked cleft palate and ankyloglossia , 2001, Nature Genetics.

[35]  K Basler,et al.  Compartment boundaries: at the edge of development. , 1999, Trends in genetics : TIG.

[36]  M. Magnuson,et al.  Polydactyly and ectopic ZPA formation in Alx-4 mutant mice. , 1997, Development.

[37]  Ruijin Huang,et al.  Origin of the epaxial and hypaxial myotome in avian embryos , 2000, Anatomy and Embryology.

[38]  N. Copeland,et al.  Gene and pseudogene of the mouse cation-dependent mannose 6-phosphate receptor. Genomic organization, expression, and chromosomal localization. , 1992, The Journal of biological chemistry.

[39]  D. Bennett,et al.  The color loci of mice--a genetic century. , 2003, Pigment cell research.

[40]  P. Lane,et al.  Gene order in linkage group XVI of the house mouse. , 1979, The Journal of heredity.

[41]  A. Meyer,et al.  Cloning of zebrafish T-box genes tbx15 and tbx18 and their expression during embryonic development , 2002, Mechanisms of Development.

[42]  C. MacArthur,et al.  Roles for FGF8 in the Induction, Initiation, and Maintenance of Chick Limb Development , 1996, Cell.

[43]  B. Dreyfus,et al.  Divergent sexual selection enhances reproductive isolation in sticklebacks , 2022 .

[44]  Randy L. Johnson,et al.  Interactions between dorsal-ventral patterning genes lmx1b, engrailed-1 and wnt-7a in the vertebrate limb. , 2002, The International journal of developmental biology.

[45]  C. Rodriguez,et al.  Dorsal cell fate specified by chick Lmx1 during vertebrate limb development , 1996, Nature.

[46]  A. Wilkie,et al.  Haploinsufficiency of the human homeobox gene ALX4 causes skull ossification defects , 2001, Nature Genetics.

[47]  A. Joyner,et al.  ABSTRACT Two lineage boundaries and En1 coordinate AER formation , 2000 .

[48]  D. Houzelstein,et al.  The expression of the homeobox gene Msx1 reveals two populations of dermal progenitor cells originating from the somites. , 2000, Development.

[49]  A. McMahon,et al.  Novel regulatory interactions revealed by studies of murine limb pattern in Wnt-7a and En-1 mutants. , 1997, Development.

[50]  D. Cox,et al.  An autosomal dominant syndrome with 'acromegaloid' features and thickened oral mucosa. , 1985, Journal of medical genetics.

[51]  L. Niswander Pattern formation: old models out on a limb , 2003, Nature Reviews Genetics.

[52]  G. Barsh,et al.  The interaction of agouti signal protein and melanocyte stimulating hormone to regulate melanin formation in mammals. , 1996, Pigment cell research.

[53]  James Mallet,et al.  Reproductive isolation caused by colour pattern mimicry , 2001, Nature.

[54]  W. Silvers,et al.  The development of regional pigmentation patterns in black and tan (at) mice. , 1976, The Journal of experimental zoology.

[55]  G. Barsh,et al.  Cloning of the mouse agouti gene predicts a secreted protein ubiquitously expressed in mice carrying the lethal yellow mutation. , 1993, Genes & development.

[56]  W. Wuyts,et al.  The ALX4 homeobox gene is mutated in patients with ossification defects of the skull (foramina parietalia permagna, OMIM 168500) , 2000, Journal of medical genetics.

[57]  J. Guénet,et al.  Mouse Alx3: an aristaless-like homeobox gene expressed during embryogenesis in ectomesenchyme and lateral plate mesoderm. , 1998, Developmental biology.

[58]  C. Beechey,et al.  Genetic regulation of alcohol dehydrogenase C2 in the mouse. Developmental consequences of the temporal locus (Adh-3t) and positioning of Adh-3 on chromosome 3 , 1981 .

[59]  An experimental approach to action of genes at the agouti locus in the mouse. III. Transplants of newborn Aw-, A-and at-skin to Ay-, Aw-, A-and aa hosts. , 1955, The Journal of experimental zoology.

[60]  Antony G. Searle,et al.  Comparative genetics of coat colour in mammals. , 1968 .

[61]  A. Burke,et al.  A new view of patterning domains in the vertebrate mesoderm. , 2003, Developmental Cell.

[62]  J. Izpisúa-Belmonte,et al.  Involvement of FGF-8 in initiation, outgrowth and patterning of the vertebrate limb. , 1996, Development.

[63]  T. Vogt,et al.  Mutations in mouse Aristaless-like4 cause Strong's luxoid polydactyly. , 1998, Development.

[64]  G. Curry Genetical and development studies on droopy-eared mice. , 1959, Journal of embryology and experimental morphology.

[65]  M. Belk,et al.  Pelage coloration in oldfield mice (Peromyscus polionotus) : Antipredator adaptation ? , 1996 .

[66]  Juan Carlos Izpisúa Belmonte,et al.  Dorsal cell fate specified by chick Lmxl during vertebrate limb development , 1995, Nature.

[67]  G. Barsh,et al.  Biochemical and genetic studies of pigment-type switching. , 2000, Pigment cell research.

[68]  D G Wilkinson,et al.  Detection of messenger RNA by in situ hybridization to tissue sections and whole mounts. , 1993, Methods in enzymology.

[69]  S. Aparício,et al.  Eomesodermin is required for mouse trophoblast development and mesoderm formation , 2000, Nature.

[70]  G. Barsh,et al.  Differences in dorsal and ventral pigmentation result from regional expression of the mouse agouti gene. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[71]  D. Hadden,et al.  An autosomal dominant syndrome of acromegaloid facial appearance and generalised hypertrichosis terminalis. , 1996, Journal of medical genetics.

[72]  Elizabeth S. Russell Comparative Genetics of Coat Colour in Mammals. A. G. Searle. Logos Press, London; Academic Press, New York, 1968. xii + 308 pp., illus. $17.50 , 1968 .

[73]  L. Silver,et al.  Phylogenetic analysis of T-Box genes demonstrates the importance of amphioxus for understanding evolution of the vertebrate genome. , 2000, Genetics.

[74]  A. Burke,et al.  The lateral somitic frontier: dorso-ventral aspects of anterio-posterior regionalization in avian embryos , 2003, Mechanisms of Development.

[75]  A. Mauger [The role of somitic mesoderm in the development of dorsal plumage in chick embryos. I. Origin, regulative capacity and determination of the plumage-forming mesoderm]. , 1972, Journal of embryology and experimental morphology.

[76]  B. Dallapiccola,et al.  Acromegaloid facial appearance (AFA) syndrome: report of a second family. , 1992, Journal of medical genetics.

[77]  M. Marks,et al.  The melanosome: membrane dynamics in black and white , 2001, Nature Reviews Molecular Cell Biology.