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Regeneration (Oxf)
2015 Mar 13;21:19-25. doi: 10.1002/reg2.27.
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Distinct patterns of endosulfatase gene expression during Xenopus laevis limb development and regeneration.
Wang YH
,
Beck C
.
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The heparan sulfate 6-O-endosulfatases sulf1 and sulf2 regulate multiple cellular processes and organ development. Sulfs modulate a range of heparan-sulfate-dependent extracellular pathways, including the fibroblast growth factor, bone morphogenetic protein, and wingless/wnt signaling pathways. Known patterns of sulf transcript expression together with functional experiments have implicated the sulfs in chondrogenesis and muscle regeneration in mammals. Here, we describe the expression patterns of Xenopus laevis sulf1 and sulf2 in developing forelimbs and hindlimbs and demonstrate novel expression of the sulf transcripts in the regenerating hindlimbs, with prominent sulf2 expression in the proliferating blastema and transient expression of sulf1 in the redeveloping apical epidermal ridge. These findings further suggest involvement of the sulfs in successful limb regeneration in amphibians.
Figure 1.
Sulf1 expression in X. laevis stage 51−56 forelimbs (A−E) and stage 51−56 hindlimbs (F−L) examined by a whole mount in situ hybridization. Specific staining is dark purple. Red arrows in the more proximal part indicate gene expression at the putative shoulder/hip and elbow/knee joints; blue arrows indicate expression at the future wrist/ankle joints; yellow arrows indicate transcript expression in the autopod region. Roman numerals indicate digit number. All limbs are oriented proximal to the left and posterior uppermost, with the tadpoles lying on their right side.
Figure 2.
Sulf2 expression in X. laevis stage 51−53 forelimbs (A−C) and stage 50−57 hindlimbs (D−J) examined by a whole mount in situ hybridization. Specific staining is dark purple. Red arrows in the more proximal part indicate gene expression at the putative shoulder/hip and elbow/knee joints; blue arrows indicate expression at the future wrist/ankle joints; yellow arrows indicate transcript expression in the autopod region; empty arrows indicate reduced level of expression. Roman numerals indicate digit number. The boxed region in E indicates area of histological section shown in E′, accompanied by the corresponding schematic (E′) showing area of gene expression in the mesenchyme. All limbs are oriented proximal to the left and posterior uppermost, with the tadpoles lying on their right side. AER, apical epidermal ridge; m, mesenchyme.
Figure 3.
X. laevis sulf1 (A−F) and sulf2 (G−L) expression in regenerating hindlimbs originally operated at developmental stage 52 examined by a whole mount in situ hybridization. Specific staining is dark purple. Black arrows point to regions of transcript expression that is specific to regeneration; empty arrows indicate reduced level of expression. The original site of amputation is indicated by the black dotted line. Boxed region in C and I indicate area of histological section as shown in C′ and I′ respectively, accompanied by their respective schematics. All limbs are oriented proximal to the left and posterior uppermost, with the tadpoles lying on their right side. aec, apical epidermal cap; fa, functional aec; b, blastema.
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