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Expression of skeletogenic genes during arm regeneration in the brittle star Amphiura filiforrnis
Czarkwiani, A.; Dylus, D.V.; Oliveri, P. (2013). Expression of skeletogenic genes during arm regeneration in the brittle star Amphiura filiforrnis. Gene expression patterns (Tokyo) 13(8): 464-472. https://dx.doi.org/10.1016/j.gep.2013.09.002
In: Gene expression patterns (Tokyo). ISSN 1567-133X; e-ISSN 1872-7298, more
Peer reviewed article  

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Keywords
    Data
    Marine Sciences
    Marine Sciences > Marine Genomics
    Scientific Community
    Scientific Publication
    Marine/Coastal
Author keywords
    Regeneration; Brittle star; Transcription factors; Skeleton; ets1/2;tbr; gataC; foxB; alx1

Project Top | Authors 
  • Association of European marine biological laboratories, more

Authors  Top 
  • Czarkwiani, A.
  • Dylus, D.V.
  • Oliveri, P.

Abstract
    The brittle star Amphiura filiformis, which regenerates its arms post autotomy, is emerging as a useful model for studying the molecular underpinnings of regeneration, aided by the recent availability of some molecular resources. During regeneration a blastema initially is formed distally to the amputation site, and then a rapid rebuild is obtained by adding metameric units, which will eventually differentiate and become fully functional. In this work we first characterize the developmental process of the regenerating arms using two differentiation markers for muscle and skeletal structures – Afi-trop-1 and Afi-αcoll. Both genes are not expressed in the blastema and newly added undifferentiated metameric units. Their expression at different regenerating stages shows an early segregation of muscle and skeletal cells during the regenerating process, long before the metameric units become functional. We then studied the expression of a set of genes orthologous of the sea urchin transcription factors involved in the development of skeletal and non-skeletal mesoderm: Afi-ets1/2, Afi-alx1, Afi-tbr, Afi-foxB and Afi-gataC. We found that Afi-ets1/2, Afi-alx1, Afi-foxB and Afi-gataC are all expressed at the blastemal stage. As regeneration progresses those genes are expressed in a similar small undifferentiated domain beneath the distal growth cap, while in more advanced metameric units they become restricted to different skeletal domains. Afi-foxB becomes expressed in non-skeletal structures. This suggests that they might play a combinatorial role only in the early cell specification process and that subsequently they function independently in the differentiation of different structures. Afi-tbr is not present in the adult arm tissue at any stage of regeneration. In situ hybridization results have been confirmed with a new strategy for quantitative PCR (QPCR), using a subdivision of the three stages of regeneration into proximal (differentiated) and distal (undifferentiated) arm segments.

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