Screen for genes differentially expressed during regeneration of the zebrafish caudal fin

Abstract

The zebrafish caudal fin constitutes an important model for studying the molecular basis of tissue regeneration. The cascade of genes induced after amputation or injury, leading to restoration of the lost fin structures, include those responsible for wound healing, blastema formation, tissue outgrowth, and patterning. We carried out a systematic study to identify genes that are up-regulated during "initiation" (1 day) and "outgrowth and differentiation" (4 days) of fin regeneration by using two complementary methods, suppression subtraction hybridization (SSH) and differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). We obtained 298 distinct genes/sequences from SSH libraries and 24 distinct genes/sequences by DDRT-PCR. We determined the expression of 54 of these genes using in situ hybridization. In parallel, gene expression analyses were done in zebrafish embryos and early larvae. The information gathered from the present study provides resources for further investigations into the molecular mechanisms of fin development and regeneration.

Characterization of two new zebrafish members of the hedgehog family: Atypical expression of a zebrafish indian hedgehog gene in skeletal elements of both endochondral and dermal origins

“…3C-E) that are intramembranous bones, devoid of cartilage precursor. Padhi et al (2004) have also reported the expression of zebrafish col10a1 in the cleithrum, another bone of intramembranous origin, between 2 and 3dpf (Padhi et al, 2004), and consistent with their observations, we observed col10a1 expression in the cleithrum at 4dpf (Fig. 3F).…”

“…In a previous study, Padhi et al isolated an EST clone (1-A06) from a 4dpa regeneration subtraction library, which presents similarities with the mammalian collagen type X alpha 1 chain [col10a1 (Padhi et al, 2004)]. We further analyzed this sequence, and comparison of 1-A06 with the zebrafish EST database (http://zfish.…”

Inhibition of zebrafish fin regeneration using in vivo electroporation of morpholinos against fgfr1 and msxb

“…Given that the efficiency of knock-down varies greatly between morpholinos, it would be difficult to argue for using this technique to compare which of two genes has a greater effect on fin outgrowth. However, as the expression of new molecules during regeneration are being identified in microarray or differential display screens, this technique does permit placing these molecules into known regeneration pathways and revealing the functional relevance of these genes for regeneration (Katogi et al, 2004;Padhi et al, 2004). For example, knock-down of Msxb did not effect msxc expression at 3 days post-amputation, but both msxb and msxc expression were greatly reduced or absent when Fgfr1 levels were knocked-down (Poss et al, 2000; Fig.…”

Tissue regeneration after injury in adult zebrafish: The regenerative potential of the caudal fin

“…Genes and cell-signaling pathways that are involved in zebrafish fin regeneration have been previously characterized (Laforest et al, 1998;Tawk et al, 2000;Quint et al, 2002;Padhi et al, 2004;Smith et al, 2006;Jazwinska et al, 2007;Stoick-Cooper et al, 2007a, b;Wills et al, 2008;Yin et al, 2008;Chablais and Jazwinska, 2010). For example, Whitehead et al (2005) reported that the expression of fgf20a, a gene that encodes a member of the fibroblast growth factor family, plays important roles in the initiation of fin regeneration, and that decreases in fgf20a expression result in inhibition of fin regeneration.…”