RESUMO
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Assuntos
Animais , Quitridiomicetos/isolamento & purificação , Anfíbios/microbiologia , Itraconazol/uso terapêutico , Microscopia Eletrônica de Varredura/métodos , Antifúngicos/uso terapêuticoRESUMO
The goal of our study was the identification of up-regulated genes during axolotl(Ambystoma mexicanum) hindlimb regeneration 4 days after amputation using suppressionsubtractive hybridization (SSH). Approximately 400 clones that harboredupregulated genes in regenerating blastema tissue were selected for sequence analysis.A BLAST homology search against NCBI non-redundant database and anambystoma EST database revealed 102 clones that showed homology to knownsequences in GenBank with annotated function, 31 were known genes withoutknown function, 74 were novel and 72 belonged to mitochondrial sequences. Differentialexpression of Hmox1, Orc4L, Pls3, Fen-1, Mcm7 and Mmp3/10a was confirmedusing qRT-PCR analysis. Among all genes, only Mmp3/10a has been previouslydescribed as involved in limb regeneration. Other important identified genesbelong to the group of cell cycle regulators (Orc4L, Nasp, Skp1A and Mcm7, the latterbeing a possible proliferative marker), those involved in protein synthesis andtransport (Sec63, Srp72, Sara2) and V- ATPase pump. The novel genes we identifiedmight be important for the process of blastema formation and the onset of cell proliferationin a regenerating limb (AU)
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