Role of smoothened and sfrp genes in Eupentacta fraudatrix regeneration

The secreted frizzled-related proteins (sfrp) and smoothened (smo) genes and their possible role in the regeneration of internal organs in the holothurian Eupentacta fraudatrix were studied. In this species, two sfrp genes were identified: sfrp1/2/5, sfrp3/4 and one smo gene. Their expression was analysed during regeneration of the aquapharyngeal bulb (AB) and intestine, and these genes were knock down by RNA interference. It has been shown that the expression of these genes is extremely important for the formation of AB. In all animals subjected to knockdown, at 7 days after evisceration, a full-sized AB rudiment was not formed. As a result of sfrp1/2/5 knockdown, the process of extracellular matrix remodelling in AB is interrupted, that leading to clusters of dense connective tissue formation, which slows down cell migration. When sfrp3/4 is knockdown, the connective tissue of AB anlage is completely disrupted and its symmetry is broken. The effect of smo knockdown was expressed in a significant impairment of AB regeneration, when connections between ambulacras were not formed after evisceration. However, despite severe disturbances in AB regeneration, a normal-sized gut anlage developed in all cases, which suggests that the regeneration of the digestive tube and AB occur independently of each other.