Synergistic interactions between bFGF and a TGF-beta family member may mediate myogenic signals from the neural tube.

Development of the myotome within somites depends on unknown signals from the neural tube. The present study tested the ability of basic fibroblast growth factor (bFGF), transforming growth factor-beta1 (TGF-beta1) and dorsalin-1 (dsl-1) to promote myogenesis in stage 10-14 chick paraxial mesoderm utilizing 72 hour explant cultures. Each of these factors alone and the combination of bFGF with dsl-1 had limited to no myogenic-promoting activity, but the combination of bFGF with TGF-beta1 demonstrated a potent dose-dependent effect. In addition, bFGF enhanced the survival/proliferation of somite cells. 98% of stage 10-11 caudal segmental plate explants treated with bFGF plus TGF-beta1, exhibited myosin heavy chain (MHC)-positive cells (avg.=60 per explant), whereas only 15% of similarly treated somites responded with an average of 5 MHC-positive cells. Thus at stage 10-11, there are rostrocaudal differences in myogenic responsiveness with the caudal (more 'immature') paraxial mesoderm being more myogenically responsive to these factors than are somites. It was also discovered that 17% of stage 10-11 caudal segmental plate explants exhibited several MHC-positive cells even when cultured without added growth factors, further demonstrating a different myogenic potential of the caudal paraxial mesoderm. Stage 13-14 paraxial mesoderm also exhibited a myogenic response to bFGF/TGF-beta1 but, unlike stage 10-11 embryos, both somites and segmental plate exhibited a strong response. A two-step mechanism for the bFGF/TGF-beta1 effect is suggested by the finding that only TGF-beta1 was required during the first 12 hours of culture, whereas bFGF plus a TGF-beta-like factor were required for the remainder of the culture. The biological relevance of the findings with bFGF is underscored by the observation that a monoclonal antibody to bFGF inhibited myogenic signaling from the dorsal neural tube. However, a monoclonal antibody that can neutralize the three factors TGF-beta1, TGF-beta2 and TGF-beta3 did not block myogenic signals from the neural tube, raising the possibility that another TGF-beta family member may be involved in vivo.