title: Expression of helix-loop-helix type negative regulators of differentiation during limb regeneration in urodeles and anurans.
abstract: The urodele is capable of regenerating its limb by forming a blastema even in the adult. By contrast, the anuran, which is phylogenetically close to the urodele, loses this ability during metamorphosis and forms blastema-like tissues that develop only into a spike-like structure in the adult. In order to compare the molecular mechanism of the formation and maintenance of the blastema between the urodele and anuran, the genes encoding helix-loop-helix (HLH) type negative regulators of differentiation were characterized for both the Japanese newt, Cynops pyrrhogaster, and African clawed frog, Xenopus laevis. Cynops homologs of Id2, Id3, and HES1 and Xenopus Id2 were identified. To learn the roles of these genes in regeneration, their expression was examined. The expression of Id2 and Id3 was low in unamputated limbs, but was up-regulated in blastemas of both adult newt and Xenopus. Interestingly, transcripts of the two Id genes showed specific localizations in the blastema and the expression patterns were very similar in both species through the early to medium bud stage. Id2 was expressed predominantly in the blastemal epidermis, and Id3 was expressed equally in the blastemal epidermis and mesenchyme including cells in precartilage condensations. HES1 expression was up-regulated in the newt blastemal epidermis. It was thought that the up-regulation of these genes in the epidermis was related to the proliferation of the cells and that increased expression of these genes in the mesenchyme was related to the undifferentiated state of the blastemal cells. These results and considerations strongly suggested that the state of differentiation is similar in the early to medium bud blastema of both urodeles and anurans. The expression of Id3 remained high through to the digits stage in newts. In contrast, its expression in Xenopus decreased in spike-like regenerates, which correspond to palette-digits stage of newt regenerates. From these results, it was suggested that the blastema redifferentiates earlier in the frog than in the newt, and therefore the timing of redifferentiation of the cartilage is crucial for complete regeneration.
authors.affiliation: Department of Biological Science, Faculty of Science, Hiroshima University, Higashihiroshima, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Shimizu-Nishikawa
authors.forename: I
authors.initials: I
authors.lastname: Tazawa
authors.forename: K
authors.initials: K
authors.lastname: Uchiyama
authors.forename: K
authors.initials: K
authors.lastname: Yoshizato
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001001:Anura / Q000378:metabolism*; D002454:Cell Differentiation; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D005121:Extremities / Q000502:physiology; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D018257:Helix-Loop-Helix Motifs / Q000235:genetics*; D017403:In Situ Hybridization; D051797:Inhibitor of Differentiation Protein 2; D051796:Inhibitor of Differentiation Proteins; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D012038:Regeneration / Q000502:physiology; D012097:Repressor Proteins; D012455:Salamandridae / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics; D014562:Caudata / Q000378:metabolism*; D014982:Xenopus laevis / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D051797:Inhibitor of Differentiation Protein 2; D051796:Inhibitor of Differentiation Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors; C075640:ID3 protein, human