Nerve-independence of limb regeneration in larval Xenopus laevis is correlated to the level of fgf-2 mRNA expression in limb tissues.
In both larval and adult urodele amphibians, limb blastema formation requires the presence of an adequate nerve supply. In previous research, we demonstrated that the hindlimb of early Xenopus laevis larvae formed a regeneration blastema even when denervated, while the denervated limb of late larvae did not. We hypothesized that the nerve-independence was due to the autonomous synthesis of a mitogenic neurotrophic-like factor by undifferentiated limb bud cells. In this paper, we demonstrate that fgf-2 mRNA is present in larval limb tissues and that its level is correlated to the extent of mesenchymal cells populating the limb: in early limbs, fgf-2 mRNA is present at high levels all over the limb, while, in late limbs, the fgf-2 expression is low and detectable only in the distal autopodium. After denervation, fgf-2 mRNA synthesis increases in amputated early limbs but not in amputated late limbs. The implantation of anti-FGF-2 beads into amputated early limbs hardly lowers the mitotic activity of blastema cells. However, FGF-2 beads implanted into the blastema of late limbs prevent the denervation-induced inhibition of mitosis and oppose blastema regression. Our data indicate that FGF-2 is a good candidate for the endogenous mitogenic factor responsible for blastema formation and growth in amputated and denervated early limbs. However, in amputated late limbs, the very limited fgf-2 expression is not sufficient to promote blastema formation in the absence of nerves.
Dipartimento di Biologia, Università di Roma "Tor Vergata", Via della Ricerca Scientifica, Rome, 00133, Italy.
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Cannata
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Bagni
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Bernardini
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Christen
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Filoni
D000818:Animals; D002455:Cell Division; D005121:Extremities / Q000294:innervation / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000378:metabolism; D017403:In Situ Hybridization; D007814:Larva / Q000502:physiology*; D008648:Mesoderm / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014982:Xenopus laevis
D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2