Anatomy and Embryology

, Volume 208, Issue 1, pp 7–18 | Cite as

Characterization of migration behavior of myogenic precursor cells in the limb bud with respect to Lmx1b expression

Original Article


Limb buds develop from lateral plate-derived stationary mesenchyme and are invaded by cells from extrinsic regions. The largest populations of these cells are myogenic precursor cells that originate from the lateral dermomyotomes. After detachment under the influence of SF/HGF, myogenic precursor cells migrate in a proximo-distal direction and populate a dorsal and ventral zone. The patterning mechanism leading to the segregation of dorsal and ventral myogenic cells is at present not understood. Lmx1b, a LIM homeodomain transcription factor expressed in the dorsal mesenchyme of the developing limb bud, forms a sharp dorso-ventral boundary of expression within the limb. We have investigated the mechanisms of dorso-ventral patterning of muscle precursor cells in the limb buds with respect to Lmx1b expression using quail-chick chimeras and transgenic mice. Although cells appeared to be capable of migrating either ventrally or dorsally, their migration was restricted to the position they had attained during normal development or in the experimental situation. They were never found to cross the dorso-ventral boundary. Immunohistochemistry and histological analysis of mice carrying a LacZ reporter gene under the control of the endogenous Lmx1b locus confirmed that myogenic precursors in the limb bud were devoid of Lmx1b expression. In addition, it was shown that Lmx1b is not only expressed at early stages of limb development but maintains its pattern, at least until after birth. The present study provides new insights into migratory pathways of myogenic precursor cells and reveals details of Lmx1b expression on a cellular basis within the limb.


Mouse Chick embryo Muscle Lbx1 Quail-chick labeling technique Cell migration Nail-Patella-Syndrome 


  1. Beddington RS, Martin P (1989) An in situ transgenic enzyme marker to monitor migration of cells in the mid-gestation mouse embryo. Somite contribution to the early forelimb bud. Mol Biol Med 6:263–274PubMedGoogle Scholar
  2. Beresford B (1983) Brachial muscles in the chick embryo: the fate of individual somites. J Embryol Exp Morphol 77:99–116PubMedGoogle Scholar
  3. Bladt F, Riethmacher D, Isenmann S, Aguzzi A, Birchmeier C (1995) Essential role for the c-met receptor in the migration of myogenic precursor cells into the limb bud. Nature 376:768–771PubMedGoogle Scholar
  4. Brand B (1987) Zur Morphogenese und Musterbildung der Vogelextremität unter besonderer Berücksichtigung der Migration der myogenene Zellen. Dissertation, BochumGoogle Scholar
  5. Brand-Saberi B, Krenn V, Christ B (1989) The control of directed myogenic cell migration in the avian limb bud. Anat Embryol (Berl) 180:555–566Google Scholar
  6. Brand-Saberi B, Krenn V (1991) Observations concerning the control of directed myogenic cell migration. In: Hinchliffe JR (ed) Developmental patterning of the vertebrate limb. Plenum, New York, pp 273–284Google Scholar
  7. Brand-Saberi B, Christ B (1992) A comparative study of myogenic cell invasion of the avian wing and leg bud. Eur J Morphol 30:169–180PubMedGoogle Scholar
  8. Brand-Saberi B, Krenn V, Grim M, Christ B (1993) Differences in the fibronectin-dependence of migrating cell populations. Anat Embryol (Berl) 187:17–26Google Scholar
  9. Brand-Saberi B, Seifert R, Grim M, Wilting J, Kuhlewein M, Christ B (1995) Blood vessel formation in the avian limb bud involves angioblastic and angiotrophic growth. Dev Dyn 202:181–194PubMedGoogle Scholar
  10. Brand-Saberi B, Gamel AJ, Krenn V, Muller TS, Wilting J, Christ B (1996A) N-cadherin is involved in myoblast migration and muscle differentiation in the avian limb bud. Dev Biol 178:160–173CrossRefPubMedGoogle Scholar
  11. Brand-Saberi B, Muller TS, Wilting J, Christ B, Birchmeier C (1996B) Scatter factor/hepatocyte growth factor (SF/HGF) induces emigration of myogenic cells at interlimb level in vivo. Dev Biol 179:303–308CrossRefPubMedGoogle Scholar
  12. Bronner-Fraser M (1986) Analysis of the early stages of trunk neural crest migration in avian embryos using monoclonal antibody HNK-1. Dev Biol 115:44–55PubMedGoogle Scholar
  13. Campeau E, Watkins D, Rouleau GA, Babul R, Buchanan JA, Meschino W, Der Kaloustian VM (1995) Linkage analysis of the nail-patella syndrome. Am J Hum Genet 56:243–247PubMedGoogle Scholar
  14. Chen H, Lun Y, Ovchinnikov D, Kokubo H, Oberg KC, Pepicelli CV, Gan L, Lee B, Johnson RL (1998A) Limb and kidney defects in Lmx1b mutant mice suggest an involvement of LMX1B in human nail patella syndrome. Nat Genet 19:51–55PubMedGoogle Scholar
  15. Chen H, Ovchinnikov D, Pressman CL, Aulehla A, Lun Y, Johnson RL (1998B) Multiple calvarial defects in lmx1b mutant mice. Dev Genet 22:314–320CrossRefPubMedGoogle Scholar
  16. Chevallier A, Kieny M, Mauger A (1977) Limb-somite relationship: origin of the limb musculature. J Embryol Exp Morphol 41:245–258PubMedGoogle Scholar
  17. Christ B, Jacob HJ, Jacob M (1974) Origin of wing musculature. Experimental studies on quail and chick embryos. Experientia 30:1446–1449PubMedGoogle Scholar
  18. Christ B, Jacob HJ, Jacob M (1977) Experimental analysis of the origin of the wing musculature in avian embryos. Anat Embryol (Berl) 150:171–186Google Scholar
  19. Christ B, Ordahl CP (1995) Early stages of chick somite development. Anat Embryol (Berl) 191:381–396Google Scholar
  20. Dealy CN, Roth A, Ferrari D, Brown AM, Kosher RA (1993) Wnt-5a and Wnt-7a are expressed in the developing chick limb bud in a manner suggesting roles in pattern formation along the proximodistal and dorsoventral axes. Mech Dev 43:175–186CrossRefPubMedGoogle Scholar
  21. Dietrich S, Schubert FR, Healy C, Sharpe PT, Lumsden A (1998) Specification of the hypaxial musculature. Development 125:2235–2249PubMedGoogle Scholar
  22. Dietrich S, Abou-Rebyeh F, Brohmann H, Bladt F, Sonnenberg-Riethmacher E, Yamaai T, Lumsden A, Brand-Saberi B, Birchmeier C (1999) The role of SF/HGF and c-Met in the development of skeletal muscle. Development 126:1621–1629PubMedGoogle Scholar
  23. Dreyer SD, Zhou G, Baldini A, Winterpacht A, Zabel B, Cole W, Johnson RL, Lee B (1998) Mutations in LMX1B cause abnormal skeletal patterning and renal dysplasia in nail patella syndrome. Nat Genet 19:47–50PubMedGoogle Scholar
  24. Dreyer SD, Morello R, German MS, Zabel B, Winterpacht A, Lunstrum GP, Horton WA, Oberg KC, Lee B (2000) LMX1B transactivation and expression in nail-patella syndrome. Hum Mol Genet 9:1067–1074PubMedGoogle Scholar
  25. German MS, Wang J, Chadwick RB, Rutter WJ (1992) Synergistic activation of the insulin gene by a LIM-homeo domain protein and a basic helix-loop-helix protein: building a functional insulin minienhancer complex. Genes Dev 6:2165–2176PubMedGoogle Scholar
  26. Grim M, Wachtler F (1991) Muscle morphogenesis in the absence of myogenic cells. Anat Embryol (Berl) 183:67–70Google Scholar
  27. Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–82Google Scholar
  28. Heymann S, Koudrova M, Arnold H, Koster M, Braun T (1996) Regulation and function of SF/HGF during migration of limb muscle precursor cells in chicken. Dev Biol 180:566–578CrossRefPubMedGoogle Scholar
  29. Hollyday M (1983) Development of motor innervation of chick limbs. Prog Clin Biol Res 110:183–193PubMedGoogle Scholar
  30. Holtzer H (1968) Induction of chondrogenesis: a concept in terms of mechanisms. In: Gleischmajer R, Billingham RE (eds) Epithelial-Mesenchymal Interactions. Williams, Baltimore, pp 152–164Google Scholar
  31. Jacob M, Christ B, Jacob HJ (1978) On the migration of myogenic stem cells into the prospective wing region of chick embryos. A scanning and transmission electron microscope study. Anat Embryol (Berl) 153:179–193Google Scholar
  32. Jaffredo T, Horwitz AF, Buck CA, Rong PM, Dieterlen-Lievre F (1988) Myoblast migration specifically inhibited in the chick embryo by grafted CSAT hybridoma cells secreting an anti-integrin antibody. Development 103:431–446PubMedGoogle Scholar
  33. Jagla K, Dolle P, Mattei MG, Jagla T, Schuhbaur B, Dretzen G, Bellard F, Bellard M (1995) Mouse Lbx1 and human LBX1 define a novel mammalian homeobox gene family related to the Drosophila lady bird genes. Mech Dev 53:345–356CrossRefPubMedGoogle Scholar
  34. Kawakami Y, Capdevila J, Buscher D, Itoh T, Rodriguez Esteban C, Izpisua Belmonte JC (2001) WNT signals control FGF-dependent limb initiation and AER induction in the chick embryo. Cell 104:891–900PubMedGoogle Scholar
  35. Lance-Jones C (1988) The effect of somite manipulation on the development of motoneuron projection patterns in the embryonic chick hindlimb. Dev Biol 126:408–419PubMedGoogle Scholar
  36. Martin GR (1998) The roles of FGFs in the early development of vertebrate limbs. Genes Dev 12:1571–1586PubMedGoogle Scholar
  37. McIntosh I, Clough MV, Schaffer AA, Puffenberger EG, Horton VK, Peters K, Abbott MH, Roig CM, Cutone S, Ozelius L, Kwiatkowski DJ, Pyeritz RE, Brown LJ, Pauli RM, McCormick MK, Francomano CA (1997) Fine mapping of the nail-patella syndrome locus at 9q34. Am J Hum Genet 60:133–142PubMedGoogle Scholar
  38. Morello R, Zhou G, Dreyer SD, Harvey SJ, Ninomiya Y, Thorner PS, Miner JH, Cole W, Winterpacht A, Zabel B, Oberg KC, Lee B (2001) Regulation of glomerular basement membrane collagen expression by LMX1B contributes to renal disease in nail patella syndrome. Nat Genet 27:205–208PubMedGoogle Scholar
  39. Nieto MA, Patel K, Wilkinson DG (1996) In situ hybridization analysis of chick embryos in whole mount and tissue sections. Methods Cell Biol 51:219–235PubMedGoogle Scholar
  40. Pardanaud L, Altmann C, Kitos P, Dieterlen-Lievre F, Buck CA (1987) Vasculogenesis in the early quail blastodisc as studied with a monoclonal antibody recognizing endothelial cells. Development 100:339–349PubMedGoogle Scholar
  41. Parr BA, Shea MJ, Vassileva G, McMahon AP (1993) Mouse Wnt genes exhibit discrete domains of expression in the early embryonic CNS and limb buds. Development 119:247–261PubMedGoogle Scholar
  42. Rickmann M, Fawcett JW, Keynes RJ (1985) The migration of neural crest cells and the growth of motor axons through the rostral half of the chick somite. J Embryol Exp Morphol 90:437–455PubMedGoogle Scholar
  43. Riddle RD, Ensini M, Nelson C, Tsuchida T, Jessell TM, Tabin C (1995) Induction of the LIM homeobox gene Lmx1 by WNT7a establishes dorsoventral pattern in the vertebrate limb. Cell 83:631–640PubMedGoogle Scholar
  44. Scaal M, Bonafede A, Dathe V, Sachs M, Cann G, Christ B, Brand-Saberi B (1999) SF/HGF is a mediator between limb patterning and muscle development. Development 126:4885–4893PubMedGoogle Scholar
  45. Schramm C, Solursh M (1990) The formation of premuscle masses during chick wing bud development. Anat Embryol (Berl) 182:235–247Google Scholar
  46. Searls RL, Janners MY (1971) The initiation of limb bud outgrowth in the embryonic chick. Dev Biol 24:198–213PubMedGoogle Scholar
  47. Serbedzija GN, Bronner-Fraser M, Fraser SE (1989) A vital dye analysis of the timing and pathways of avian trunk neural crest cell migration. Development 106:809–816PubMedGoogle Scholar
  48. Smidt MP, Asbreuk CH, Cox JJ, Chen H, Johnson RL, Burbach JP (2000) A second independent pathway for development of mesencephalic dopaminergic neurons requires Lmx1b. Nat Neurosci 3:337–341CrossRefPubMedGoogle Scholar
  49. Solursh M, Drake C, Meier S (1987) The migration of myogenic cells from the somites at the wing level in avian embryos. Dev Biol 121:389–396PubMedGoogle Scholar
  50. Sweeney E, Fryer A, Mountford R, Green A, McIntosh I (2003) Nail patella syndrome: a review of the phenotype aided by developmental biology. J Med Genet 40:153–162CrossRefPubMedGoogle Scholar
  51. Toole BP, Goldberg RL, Chi-Rosso G, Underhill CB, Orkin RW (1984) Hyaluronate cell interactions. In: Liss A (ed) The Role of Extracellular Matrix in Development, New YorkGoogle Scholar
  52. Vogel A, Rodriguez C, Warnken W, Izpisua Belmonte JC (1995) Dorsal cell fate specified by chick Lmx1 during vertebrate limb development. Nature 378:716–720PubMedGoogle Scholar
  53. Wilting J, Brand-Saberi B, Huang R, Zhi Q, Kontges G, Ordahl CP, Christ B (1995) Angiogenic potential of the avian somite. Dev Dyn 202:165–171PubMedGoogle Scholar
  54. Wilting J, Eichmann A, Christ B (1997) Expression of the avian VEGF receptor homologues Quek 1 and Quek 2 in blood-vascular and lymphatic endothelial and non-endothelial cells during quail embryonic development. Cell Tissue Res 288:207–223PubMedGoogle Scholar
  55. Yuan S, Schoenwolf GC (1999) The spatial and temporal pattern of C-Lmx1 expression in the neuroectoderm during chick neurulation. Mech Dev 88:243–247CrossRefPubMedGoogle Scholar
  56. Zhi Q, Huang R, Christ B, Brand-Saberi B (1996) Participation of individual brachial somites in skeletal muscles of the avian distal wing. Anat Embryol (Berl) 194:327–339Google Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Institute of Anatomy and Cell BiologyUniversity of FreiburgFreiburgGermany
  2. 2.Department of Biochemistry and Molecular BiologyUT M.D. Anderson Cancer CenterHoustonUSA

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