Electroporation into the Limb: Beyond Misexpression
Limb development has been studied for over 100 years by several generations of developmental biologists. The developing limb is one of the best models with which to study pattern formation in vertebrates. We have used chick limb development to answer a simple but basic question, namely, why heterogeneous tissues are formed at correct positions and times from a homogeneous population of cells (Pearse & Tabin, 1998).
Limb development starts as two pairs of tissue bulges in the lateral plate meso-derm (LPM). These are called the forelimb and hindlimb fields (Fig. 9.1). After limb initiation, one can clearly identify three-dimensional axes in the limb buds: the proximal-distal (PD; from shoulder to fingers), dorso-ventral (DV; from back to palm), and antero-posterior (AP; from thumb to little fingers) axes. Morphological changes and differences along these three axes are determined by pattern formation during limb bud stages. Following establishment of these axes, one can visually recognize condensation of cartilages. Muscles, tendons, and neurons migrate and differentiate after cartilage formation. Because the stages and events are easily recognized morphologically and in detail, it is therefore the limb bud is an excellent model with which to study the molecular mechanisms of embryonic patterning and tissue differentiation in vertebrates.
KeywordsLimb Development Lateral Plate Mesoderm Chick Limb Vitelline Artery pCAGGS Vector
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- Bendall, A. J., Ding, J., Hu, G., Shen, M. M., Abate-Shen, C. (1999) Msx1 antagonizes the myo-genic activity of Pax3 in migrating limb muscle precursors. Development 126, 4965–4976.Google Scholar
- Kawakami, Y., Rodríguez-León, J., Koth, C. M., Büscher, D., Itoh, T., Raya, A., Ng, J. K., Esteban, C. R., Takahashi, S., Henrique, D., Schwarz, M. F., Asahara, H., Izpisúa-Belmonte, J. C. (2003) MKP3 mediates the cellular response to FGF8 signalling in the vertebrate limb. Nat Cell Biol 5, 499–501.CrossRefGoogle Scholar
- Logan, C., Hornbruch, A., Campbell, I., Lumsden, A. (1997) The role of Engrailed in establishing the dorsoventral axis of the chick limb. Development 124, 2317–2324.Google Scholar
- Ohuchi, H., Nakagawa, T., Yamamoto, A., Araga, A., Ohata, T., Ishimaru, Y., Yoshioka, H., Kuwana, T., Nohno, T., Yamasaki, M., Itoh, N., Noji, S. (1997) The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor. Development 124, 2235–2244.Google Scholar
- Saito, D., Yonei-Tamura, S., Kano, K., Ide, H., Tamura, K. (2002) Specification and determination of limb identity: evidence for inhibitory regulation of Tbx gene expression. Development 129, 211–220.Google Scholar
- Saunders, J. W. Jr., Gasseling, M. T. (1968) “Epithelial-Mesenchymal Interractions” Fleischmajer, R., Billingham, R. F. eds., Williams and Wilkins, Baltimore, pp. 78–97.Google Scholar