Fat4-Dchs1 regulation of musculoskeletal morphogenesis and differentiation
This research aims to understand the development of the musculoskeletal system, the signalling pathways that control differentiation of muscle and skeletal cells and the factors that control skeletal shape. The lab uses genetic mouse models, cell culture and CT analysis to determine how Fat4-Dchs1 regulate musculoskeletal development.
Image 1: (A,B) Alizarin red and alcian blue staining of a wildtype (A) and Dchs1 mutant (B) sternum, which is wider and shorter. We are interested in why the sternum shape is different. (C) A microCT scan of a wildtype mouse skull. CT analysis can be used to identify changes in bone development and differentiation. (D) Muscle cells in culture: the nucleus is shown in blue and the muscle protein, myogenin, is shown by the red staining. We can use these techniques to show how muscle cell proliferation and differentiation is regulated.
Fat4-Dchs1 regulation of branching morphogenesis
This research aims to understand the signalling interactions that control branching morphogenesis during embryonic development. We use genetic mouse models, organ culture, OPT and confocal imaging analysis to determine the role of Fat4-Dchs1.
Image 2 : Branching of the ureteric tubules shown by the green (E-cadherin) staining in (A) wildtype and (B) Fat4 mutants. In the Fat4 mutants there is decreased branching and the epithelium is dysmorphic (arrow). See Mao et al. Development (2011) 138:947-57 for further details.
Fat4-Dchs1 regulation of neuronal migration
Collective neuronal migrations contribute significantly to the architecture of the brain. Defects in neuronal migrations are also associated with a number of neuronal disorders ranging from lissencephaly, where the ordered arrangement of neuronal layers fails to occur, to the neuropsychiatric spectrum of disorders. The aim of this research is to understand how Fat4-Dchs1 signalling regulates the collective polarised migration of neurons to their final destination. The research uses genetic mouse models, live imaging to understand when and how Fat4-Dchs1 control neuronal development.
Image 3 : Migration of the facial branchiomotor neurons (shown in blue) in (A) wildtype and (B) Fat4 mutants. In wildtype embryos, these neurons arise within r4 of the hindbrain and first migrate caudally (c) into r5 and r6 and then turn laterally (l). In the Fat4 mutants the neurons can migrate caudally but can not migrate laterally.The red staining in panel (A) shows Fat4 expression which is highest laterally. This polarised expression of Fat4 guides the neurons so that they turn laterally. See Zakaria et al. (2014) Current Biology 24: 1620-7 for further details.
Mechanoregulation of mesenchymal stem cell differentiation and embryonic cell fate
The aim of this research is to determine how mechanical cues such as substrate stiffness, cell density and surface area influence cell fate decisions through changes in transcriptional effectors and the actin cytoskeleton. The research uses genetic mouse models, tissue culture, engineering of scaffolds and atomic force microscopy.
Image 4. The shape and actin cytoskeleton (red staining) of mesenchymal stem cells is determined by the matrix stiffness. In (A) cells have been plated on a soft matrix and are small and round whereas in (B) cells have been plated on a stiff matrix and are large. In turn, this differential response to matrix stiffness determines the localisation of the transcriptional effector, Yap (shown in green) and ultimately cell fate: cells form adipocytes (fat cells) on soft matrix when Yap is localised to the cytoplasm and form bone cells on a stiff matrix when Yap is localised to the nucleus.
• Abu-Elmagd, M., Robson, L., Sweetman, D., Hadley, J., Francis-West, P.H*. and Münsterberg, A.* (2010) Wnt/Lef1 signalling acts via Pitx2 to regulate somite myogenesis. Dev. Biol. 337: 211-9 * equal contributions
•Nurminsky D, Shanmugasundaram S, Deasey S, Michaud C, Allen S, Hendig D, Dastjerdi A, Francis-West P, Nurminskaya M. (2011) Transglutaminase 2 regulates early chondrogenesis and glycosaminoglycan synthesis. Mech Dev. 128(3-4):234-45
• Mao Y, Mulvaney J, Zakaria S, Yu T, Morgan KM, Allen S, Basson MA, Francis-West P*, Irvine KD. * (2011) Characterisation of a Dchs1 mutant mouse reveals requirements for Dchs1-Fat4 signalling during mammalian development. Development. 2011 138:947-57.* equal contributions
• Balczerski B, Zakaria S, Tucker AS, Borycki AG, Koyama E, Pacifici M, Francis-West P. (2012) Distinct spatiotemporal roles of hedgehog signalling during chick and mouse cranial base and axial skeleton development. Dev Biol.;371(2):203-14
• Kobayashi GS, Alvizi L, Sunaga DY, Francis-West P, Kuta A, Almada BV, Ferreira SG, de Andrade-Lima LC, Bueno DF, Raposo-Amaral CE, Menck CF, Passos-Bueno MR. (2013) Susceptibility to DNA damage as a molecular mechanism for non-syndromic cleft lip and palate. PLoS One. 2013 Jun 12;8(6):e65677
•Mullen L, Rigby A, Sclanders M, Adams G, Mittal G, Colston J, Fatah R, Subang C, Foster J, Francis-West P, Köster M, Hauser H, Layward L, Vessillier S, Annenkov A, Al-Izki S, Pryce G, Bolton C, Baker D, Gould DJ, Chernajovsky Y. (2014) Latency can be conferred to a variety of cytokines by fusion with latency-associated peptide from TGF-β. Expert Opin Drug Deliv. 2014 Jan;11(1):5-16
•Zakaria S, Mao Y, Kuta A, Ferreira de Sousa C, Gaufo GO, McNeill H, Hindges R, Guthrie S, Irvine KD, Francis-West PH. (2014) Regulation of Neuronal Migration by Dchs1-Fat4 Planar Cell Polarity. Current Biology 24: 1620-7
Francis-West P.H, Hill, B. (2008) Perspective: Uncoupling the Role of Sonic Hedgehog in Limb Development: Growth and Specification, Science Signalling –1, p34
Schoenwolf, G.C., Bleyl, S.B., Brauer, P.R., Francis-West, P.H. Larsen’s “Human Embryology” 5th edition 2014. Elsevier Churchill Livingstone Press.