Organismal Dynamics

Embriología molecular: Genómica funcional, transgénesis de ratón y formación de músculo esquelético

Dr Jaime Carvajal

Investigador/a Principal

Resumen

 Unravelling the mechanisms by which a cell adopts the correct fate in response to its local environment remains one of the fundamental goals of biological research. Skeletal muscle is a paradigm for the study of cellular commitment, differentiation and organogenesis during both embryogenesis and regeneration and repair in the adult. It has long been the subject of developmental studies because it is easily recognizable from an early stage, a factor vitally important prior to the advent of molecular markers. Moreover, the establishment of myogenic cell lines has allowed its biochemistry to be studied intensively for over 30 years. In the 1990s, the use of transgenesis, new cell labelling techniques and molecular markers allowed the merging of these two approaches to provide the basis of an understanding of the molecular embryology of the system. We are now entering a new phase, in which integration of molecular embryology, comparative genomics, functional genomics and studies of transcriptional regulation will result in precise understanding of the mechanisms of determination and differentiation in the embryo.

 

Skeletal myogenesis is initiated by the activation of the Mrf4/Myf5 locus which is expressed in multiple regions of the embryo in response to signals which are interpreted by an exceptionally complex array of regulatory elements. This triggers a cascade of transcription factors and downstream structural genes, ultimately resulting in the generation of one of the fundamental histotypes. Our work will provide specific answers to a number of outstanding questions in developmental biology, such an in-depth study of the molecular biology of craniofacial muscle development which has been little characterised in mammals, transcriptional regulation, including the full characterization of particular elements involved in crucial regulatory aspects of the locus and the analysis of a novel mechanism for regulating complex loci. Finally, understanding the upstream signalling systems will open the way to studies of molecular evolution and of regeneration and repair in the adult.

Cinco publicaciones relevantes

  • Moncaut, N., Rigby, P. W. J., Carvajal, J. J. (2013) Dial M(RF) for myogenesis. FEBS J., 280: 3980-3990.
  • Schraml, B. U., van Blijswijk, J., Zelenay, S., Whitney, P. G., Acton, S. E., Filby, A., Rogers, N. C., Moncaut, N., Carvajal, J. J., Reis e Sousa, C. (2013) A model for lineage tracing of dendritic cells in mice. Cell, 154: 843-858.
  • Moncaut, N., Cross, J. W., Siligan, C., Keith, A., Taylor, K., Rigby, P. W. J., Carvajal, J. J. (2012) Musculin and TCF21 coordinate the maintenance of myogenic regulatory factor expression levels during mouse craniofacial development. Development, 139: 958-67.
  • Ribas, R., Moncaut, N., Siligan, C., Taylor, K., Cross, J. W., Rigby, P. W. J., Carvajal, J. J. (2011) Members of the TEAD family of transcription factors regulate the expression of Myf5 in ventral somitic compartments. Dev. Biol. 355: 372-380.
  • Carvajal, J. J., Keith, A., and Rigby, P. W. J. (2008) Global transcriptional regulation of the locus encoding the skeletal muscle determination genes Mrf4 and Myf5. Genes Dev., 22: 265-276.

Miembros del laboratorio y colaboradores

Miembros del laboratorio

 

Investigador Principal

Dr Jaime J. Carvajal, +34 954 978210, j.carvajal@csic.es

Postdoctoral

Cristina Vicente García 

Estudiantes de Doctorado

Macarena López Mayorga
Alejandra Navarro Martínez 

Estudiantes de Master

Pedro J. García Murillo 

Alumnos Internos

Paula Partínez López
Manuel Munuera 

Técnicos de Laboratorio

Jorge Corbach

Colaboradores Externos

Prof. Peter W.J. Rigby  (The Institute of Cancer research, London, UK)
Dr. Louis Chesler (The Institute of Cancer research, London, UK)
Prof. Richard Houlston (The Institute of Cancer research, London, UK)
Prof. Ian Tomlinson (Wellcome Trust Centre for Human Genetics, Oxford, UK)
Dr. Lydia Teboul (MRC Harwell, Oxford, UK)
Prof. Stefano Chiaffino (Padova University, Italy)
Prof. Michael Rudnicki (Ottawa University, Canada)
Prof. Thomas Braun (Max-Planck Institute, Bad Nauheim, Germany)
Dr. Anne-Gaelle Borycki (Sheffield University, UK)
Dr Gerard Grosveld (St Jude Children’s Research Hospital, Memphis, Tennessee, USA)

Lista completa de publicaciones

  1. Vicente-García, C; Villarejo_Balcells, B; Irastorza-Azcárate. I; Naranjo, S; Acemel, R. D.; Tena, J. J.; Rigby, P. W. J.; Devos, D. P.; Gómez-Skarmeta, J. L.; Carvajal, J. J. Regulatory landscape fusion in rhabdomyosarcoma through interactions between the PAX3 promoter and FOXO1 regulatory elements. Genome Biology (In Press)
  2. Moretti, I; Ciciliot, S; Dyar, K A; Abraham, R; Murgia, M; Agatea, L; Akimoto, T; Bicciato, S; Forcato, M; Pierre, P; Uhlenhaut, N H; Rigby, P W J; Carvajal, J J; Blaauw, G; Calabria, E; Schiaffino, S. (2016) MRF4 negatively regulates adult skeletal muscle growth by repressing MEF2 activity. Nature Comm. 7: 12397.
  3. Naldaiz-Gastesi, N.; Goicoechea, M.; Alonso-Martín, S.; Aiastui, A.; López-Mayorga, M.; García-Belda, P.; Lacalle, J.; San José, C.; Araúzo-Bravo, M. J.; Trouilh, L.; Anton-Leberre, V.; Herrero, D.; Matheu, A.; Bernad, A.; García-Verdugo, J. M.; Carvajal, J. J.; Relaix, F.; Lopez de Munain, A.; García-Parra, P.; Izeta, A. (2016) Identification and Characterization of the Dermal Panniculus Carnosus Muscle Stem Cells. Stem Cell Reports. 7: 411-24.
  4. Muñoz-Cánoves, P.; Carvajal, J. J.; Lopez de Munain, A.; Izeta, A. (2016) Editorial: Role of Stem Cells in Skeletal Muscle Development, Regeneration, Repair, Aging, and Disease. Front Aging Neurosci. 8: 95.
  5. Gómez-Marín, C.; Tena, J.J.; Acemel, R.D.; López-Mayorga, M.; Naranjo, S.; de la Calle-Mustiens, E.; Maeso, I.; Beccari, L.; Aneas, I.; Vielmas, E.; Bovolenta, P.; Nobrega, M.; Carvajal, J.J.; Gómez-Skarmeta, J.L. (2015) Evolutionary comparison reveals that diverging CTCF sites are signatures of ancestral topological associating domains borders. Proc Natl Acad Sci USA. 112: 7542-7.
  6. Lagutina I, Valentine V, Picchione F, Valentine MB, Villarejo-Balcells B, Carvajal JJ, Grosveld GC. Modeling of the human alveolar rhabdomyosarcoma Pax3–Foxo1 chromosome translocation in mouse myoblasts using CRISPR-Cas9 nuclease, (2015) PLOS Genetics, 11: e1004951.
  7. Lewis, A.; Freeman-Mills, L.; de la Calle-Mustienes, E.; Giráldez-Pérez, R. M.; Davis, H.; Jaeger, E.; Becker, M.; Hubner, N.; Nguyen, L.; Zeron-Medina, J.; Bond, G.; Stunnenberg, H.; Carvajal, J. J.; Skarmeta, J. L.; Leedham, S.; Tomlinson, I.A Polymorphic Enhancer near GREM1 Influences Bowel Cancer Risk through Differential CDX2 and TCF7L2 Binding. (2014) Cell Reports, 8:  983 – 990.
  8. Tena, J. J., González-Aguilera, C., Fernández-Miñán, A., Parra-Acero, H., Cross, J. W., Rigby, P. W. J., Carvajal, J. J., Wittbrodt, J., Gómez-Skarmeta, J. L., Martínez-Morales, J. R. Comparative epigenomics in distantly related teleost species identifies conserved cis- regulatory nodes active during the vertebrate phylotypic period, (2014) Genome Research, 24: 1075-1085.
  9. Bessa, J., Luengo, M., Rivero-Gil, S., Ariza-Cosano, A., Maia, A. H. F., Ruiz-Ruano, F. J., Caballero, P., Naranjo, S., Carvajal, J. J. and Gómez-Skarmeta, J. L. A mobile insulator system to detect and disrupt cis-regulatory landscapes in vertebrates,(2014) Genome Research, 24: 487-95.
  10. Schraml, B. U., van Blijswijk, J., Zelenay, S., Whitney, P. G., Acton, S. E., Filby, A., Rogers, N. C., Moncaut, N., Carvajal, J. J., Reis e Sousa, C. (2013) A model for lineage tracing of dendritic cells in mice. Cell, 154: 843-858.
  11. Moncaut, N., Rigby, P. W. J., Carvajal, J. J. (2013) Dial M(RF) for myogenesis. FEBS J., 280: 3980-3990.
  12. Harel, I., Maezawa, Y., Avraham, R., Rinon, A., Ma, H., Cross, J. W., Leviatan, N., Hegesh, J. T., Roy, A., Jacob-Hirsch, J., Rechavi, G., Carvajal, J. J., Tole, S., Kioussi, C., Quaggin, S., and Tzahor, E. (2012) Pharyngeal mesoderm regulatory network controls cardiac and head muscle morphogenesis. Proc Natl Acad Sci U S A, 109: 18839-44.
  13. Soleimani, V. D., Punch, V. G., Kawabe, Y., Jones, A. E., Carvajal, J. J., Palidwor, G., Porter, C. J., Kockx, C. E. M., van IJcken, W. F. J., Perkins, T. J., Rigby, P. W. J., Grosveld, F., and Rudnicki, M. A. (2012) Transcriptional dominance of Pax7 in adult myogenesis is due to high-affinity recognition of homeodomain motifs. Developmental Cell, 22: 1208-1220.
  14. Moncaut, N., Cross, J. W., Siligan, C., Keith, A., Taylor, K., Rigby, P. W. J., Carvajal, J. J. (2012) Musculin and TCF21 coordinate the maintenance of myogenic regulatory factor expression levels during mouse craniofacial development. Development, 139: 958-67.
  15. Ribas, R., Moncaut, N., Siligan, C., Taylor, K., Cross, J. W., Rigby, P. W. J., Carvajal, J. J. (2011) Members of the TEAD family of transcription factors regulate the expression of Myf5 in ventral somitic compartments. Dev. Biol. 355: 372-380.
  16. Villarejo-Balcells, B., Rigby, P. W. J., and Carvajal, J. J. (2011) Expression pattern of the FoxO1 gene during embryonic development. Gene Expr. Patterns 11: 299-308.
  17. Coy, S., Caamaño, J. H., Carvajal, J. J., Cleary, M.L., and Borycki A. G. (2011) A novel Gli3 enhancer controls the Gli3 spatiotemporal expression pattern through a TALE homeodomain protein binding site. Mol. Cell. Biol. 31: 1432-1443.
  18. Carvajal, J. J., and Rigby, P. W. J. (2010) Regulation of gene expression in vertebrate skeletal muscle. Exp. Cell Res. 316: 3014-3018
  19. Palles, C., Johnson, N., Coupland, B., Taylor, C., Carvajal, J., Holly, J., Fentiman, I., dos Santos Silva, I., Ashworth, A., Peto, J., and Fletcher, O. (2008) Identification of genetic variants that influence circulating IGF1 levels: a targeted search strategy. Hum. Mol. Genet. 17: 1457-1464.
  20. Carvajal, J. J., Keith, A., and Rigby, P. W. J. (2008) Global transcriptional regulation of the locus encoding the skeletal muscle determination genes Mrf4 and Myf5. Genes Dev., 22: 265-276.
  21. Hinits, Y., Osborn, D. P., Carvajal, J. J., Rigby, P. W., and Hughes, S. M. (2007) Mrf4 (myf6) is dynamically expressed in differentiated zebrafish skeletal muscle. Gene Expr. Patterns, 7: 738-745.
  22. Oustanina, S., Carvajal, J. J., Braun, T. (2007) The myogenic factor Myf5 supports efficient skeletal muscle regeneration by enabling transient myoblast amplification. Stem Cells, 25: 2006 -2016.
  23. Zammit, P. S., Carvajal, J. J., Golding, J. P., Morgan, J. E., Summerbell, D., Partridge, T. A., Rigby, P. W. J. and Beauchamp, J. R. (2004) Myf5 expression in adult muscle spindles and satellite cells is controlled by separate genetic elements. Dev Biol, 273: 454-465.
  24. Hadchouel, J., Carvajal, J. J., Daubas, P., Bajard, L., Rocancourt, D., Cox, D., Summerbell, D., Tajbakhsh, S., Rigby, P. W. J. and Buckingham, M. (2003) Analysis of a key regulatory region upstream of the Myf5 gene reveals multiple phases of myogenesis, orchestrated at each site by elements dispersed throughout the locus.Development, 130: 3415-3426.
  25. Carvajal, J. J., Cox, D., Summerbell, D. and Rigby, P. W. J. (2001) A BAC transgenic analysis of the Mrf4 / Myf5 locus reveals interdigitated elements which control activation and maintenance of gene expression during muscle development. Development 128: 1857-1868.
  26. Carvajal, J. J., Cox, D., Summerbell, D. and Rigby, P. W. J. (2001) Control of the expression of the Mrf4 and Myf5 genes: a BAC transgenic approach. Int. J. Dev Biol. 45: S139-S140
  27. Schneider, H., Groves, M., Mühle, C., Reynolds, P., Knight, A., Themis, M., Carvajal, J., Scaravilli, F., Curiel, D., Fairweather, N. and Coutelle, C. (2000) Retargeting of Adenoviral vectors to neurons using the Hc fragment of tetanus toxin. Gene Therapy 7: 1584-1592
  28. Carvajal, J., Knight, A., Schneider, H., Coutelle, C., Chamberlain S. and Fairweather, N. (1999) Non-viral neuronal gene delivery mediated by the Hc fragment of tetanus toxin. Eur. J. Biochem 259: 762-769.
  29. Pook, M.A., Carvajal, J., Doudney, K., Hillermann, R. and Chamberlain, S. (1997) Exon-intron structure of a 2.7kb transcript of the STM7 gene with phosphatidylinositol-4-phosphate 5-kinase activity. Genomics 42: 170-172.
  30. Carvajal, J., Pook, M. A., Dos Santos, M., Doudney, K., Hillermann, R., Minogue, S., Williamson, R., Hsuan, J. and Chamberlain, S. (1996) The Friedreich’s ataxia gene encodes a novel phosphatidylinositol-4-phosphate 5-kinase. Nature Genet. 14: 157-162
  31. Hillermann, R., Chee G. S., Pook, M.A., Wilkes, D., Carvajal, J., Doudney, K., Williamson, R. and Chamberlain, S. (1995) Physical evidence for the position of the Friedreich’s ataxia locus proximal to D9S5. Cytogenet Cell Genet. 71: 214-216
  32. Carvajal, J., Pook, M.A., Doudney, K., Hillermann, R., Wilkes, D., Al-Mahdawi, S., Williamson, R. and Chamberlain, S. (1995) Friedreich’s ataxia: a defect in signal transduction? Hum Mol Genet. 4: 1411-1419
  33. Carvajal, J. DNA radio-labelling and detection. In: Gene Probes: A Practical Approach.(B. D. Hames and S. Higgins, ed.). IRL Press, Oxford, 1995.
  34. Chamberlain, S., Farrall, M., Shaw, J., Wilkes, D., Carvajal, J., Doudney, K., Hillermann, R., Harding, A.E. and Williamson, R. (1993) Genetic recombination events which position the Friedreich’s ataxia locus proximal to the D9S15/D9S5 linkage group on chromosome 9q. Am J Hum Genet. 52: 99-109.
  35. Chamberlain, S., Shaw, J., Wilkes, D., Carvajal, J., Doudney, K., Hillermann, R. and Williamson, R. Molecular analysis of the Friedreich’s ataxia locus. In: Ataxias y paraplejias hereditarias. IX congreso nacional de neurología. Santander. Ediciones Ergon, 1993.
  36. Chamberlain, S., Shaw, J., Wilkes, D., Wallis, J., Carvajal, J., Fujita, R., Mandel, J.L., Farrall, M. and Williamson, R. (1991) Recombination events positioning the Friedreich’s ataxia locus in close proximity to D9S15. Cytogenet Cell Genet. 58: 3-4 (27514).