Comparative analysis of genes expressed in regenerating intestine and non-eviscerated intestine of Apostichopus japonicus Selenka (Aspidochirotida: Stichopodidae) and cloning of ependymin gene

Abstract

The regeneration of the intestine of sea cucumber (Apostichopus japonicus) was studied by describing historically the changes that occurred during intestine regeneration on the fifth day after chemicallyinduced evisceration. An expressed sequence tag (EST) analysis was undertaken to identify major genes, which might be involved in intestine regeneration of A. japonicus. Two cDNA libraries were constructed with directional cloning method, one for regenerating intestine collected on the third, fourth and fifth day after evisceration (post-evisceration, PE), and the other for the non-eviscerated (NE). A total of 730 ESTs were generated by sequencing cDNA clones from the two libraries (372 from PE and 358 from NE). The results showed that the number of genes that were involved in primary metabolism of PE library was less than that of NE library, while the number of genes involved in cell defense/immunity, cell division, cell signal transduction/communication of PE library was more than that of NE library. The results also revealed that the expression of the genes which might be involved in regeneration was enhanced to some extent after evisceration. Only about 11.54% of the sequenced clones were shared by two libraries, which provided some clues for the existence of differential gene expression between PE and NE intestines. A gene named epenAj was also characterized in this study.

Characterization and comparison of proteomes of albino sea cucumber Apostichopus japonicus (Selenka) by iTRAQ analysis

“…japonicas) [68]. In the current study, both C3 and C3-2 in albino intestine were found to be down-regulated.…”

RNA-Seq Reveals Dynamic Changes of Gene Expression in Key Stages of Intestine Regeneration in the Sea Cucumber Apostichopus japonicas

“…NADH dehydrogenase was taken as a reference gene to normalize gene expression data, based on a previous investigation [7], [16], [22], [30]. All primers were tested to be effective on PCR amplication.…”

“…Thermal cycling was as follows: (1) 95°C for 5 s; (2) 40 cycles at 95°C for 10 s, 60°C for 20 s and 72°C for 30 s. The melting curve analysis of the amplification products was done to demonstrate the specificity of the PCR products. NADH dehydrogenase (NADHF, NADHR, Table S1) was used as a housekeeping gene for internal standardization [7], [16], [22], [30]. The 2−△△CT method was used to analyze the expression level.…”

Cloning and expression analysis of Wnt6 and Hox6 during intestinal regeneration in the sea cucumber Apostichopus japonicus

“…Therefore, many genes show differential expression during intestinal regeneration (Ortiz-Pineda et al, 2009). However, only a few genes playing important roles in intestinal regeneration in sea cucumbers have been thoroughly studied, such as Wnt9, Bmp1, ependymin, serum amyloid A, survivin, and mortalin (Santiago-Cardona et al, 2003;Suarez-Castillo et al, 2004;Zheng et al, 2006;Mashanov et al, 2010Mashanov et al, , 2012. Therefore, the molecular and genetic basis of intestinal regeneration still needs further study.…”