A highly upregulated gene during tail regeneration in lizards is Wnt2b, a gene broadly expressed during development. The present study examines the distribution of Wnt proteins, most likely wnt2b, by western blotting and immunofluorescence in the blastema‐cone of lizards using a specific antibody produced against a lizard Wnt2b protein. Immunopositive bands at 48–50 and 18 kDa are present in the regenerative blastema, the latter likely as a degradation product. Immunofluorescence is mainly observed in the wound epidermis, including in the Apical Epidermal Peg where the protein appears localized in intermediate and differentiating keratinocytes. Labeling is more intense along the perimeter of keratinocytes, possibly as a secretory product, and indicates that the high epidermal proliferation of the regenerating epidermis is sustained by Wnt proteins. The regenerating spinal cord forms an ependymal tube within the blastema and shows immunolabeling especially in the cytoplasm of ependymal cells contacting the central canal where some secretion might occur. Also, regenerating nerves and proximal spinal ganglia innervating the regenerating blastema contain this signaling protein. In contrast, the blastema mesenchyme, muscles and cartilage show weak immunolabeling that tends to disappear in tissues located in more proximal regions, close to the original tail. However, a distal to proximal gradient of Wnt proteins was not detected. The present study supports the hypothesis that Wnt proteins, in particular Wnt2b, are secreted by the apical epidermis covering the blastema and released into the mesenchyme where they stimulate cell multiplication.

中文翻译：

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Wnts 在蜥蜴胚质中的免疫定位支持这些信号蛋白在尾部再生中的关键作用

蜥蜴尾部再生过程中高度上调的基因是 Wnt2b，一种在发育过程中广泛表达的基因。本研究使用针对蜥蜴 Wnt2b 蛋白产生的特异性抗体，通过西方印迹和免疫荧光法检测了 Wnt 蛋白的分布，最有可能是 wnt2b。再生胚泡中存在 48-50 和 18 kDa 的免疫阳性条带，后者可能是降解产物。免疫荧光主要在伤口表皮中观察到，包括在顶端表皮 Peg 中，其中蛋白质出现在中间和分化的角质形成细胞中。沿着角质形成细胞周边的标记更强烈，可能作为分泌产物，表明再生表皮的高表皮增殖是由 Wnt 蛋白维持的。再生脊髓在胚泡内形成室管膜管并显示免疫标记，特别是在接触可能发生一些分泌的中央管的室管膜细胞的细胞质中。此外，再生神经和支配再生胚泡的近端脊神经节也含有这种信号蛋白。相比之下，胚泡间充质、肌肉和软骨显示出弱的免疫标记，这些标记往往会在位于更近端区域的组织中消失，靠近原始尾部。然而，未检测到 Wnt 蛋白从远端到近端的梯度。本研究支持 Wnt 蛋白，特别是 Wnt2b 的假设，由覆盖胚泡的顶端表皮分泌并释放到间充质中，在那里它们刺激细胞增殖。