title: Characterization of proteolytic activities during intestinal regeneration of the sea cucumber, Holothuria glaberrima.
abstract: Proteolysis carried out by different proteases control cellular processes during development and regeneration. Here we investigated the function of the proteasome and other proteases in the process of intestinal regeneration using as a model the sea cucumber Holothuria glaberrima. This echinoderm possesses the ability to regenerate its viscera after a process of evisceration. Enzymatic activity assays showed that intestinal extracts at different stages of regeneration possessed chymotrypsin-like activity. This activity was inhibited by i) MG132, a reversible inhibitor of chymotrypsin and peptidylglutamyl peptidase hydrolase (PGPH) activities of the proteasome, ii) E64d, a permeable inhibitor of cysteine proteases and iii)  TPCK, a serine chymotrypsin inhibitor, but not by epoxomicin, an irreversible and potent inhibitor of all enzymatic activities of the proteasome. To elucidate the role which these proteases might play during intestinal regeneration, we carried out in vivo experiments injecting MG132, E64d and TPCK into regenerating animals. The results showed effects on the size of the regenerating intestine, cell proliferation and collagen degradation. These findings suggest that proteolysis by several proteases is important in the regulation of intestinal regeneration in H. glaberrima.
authors: Millenium Nucleus in Regenerative Biology (MINREB), Pontificia Universidad Católica de Chile, Chile.
authors: Consuelo
authors: C
authors: Pasten
authors: Rey
authors: R
authors: Rosa
authors: Stephanie
authors: S
authors: Ortiz
authors: Sebastián
authors: S
authors: González
authors: José E
authors: JE
authors: García-Arrarás
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D047332:Holothuria / Q000187:drug effects / Q000502:physiology*; D007422:Intestines / Q000187:drug effects / Q000502:physiology*; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D038081:Organogenesis / Q000187:drug effects / Q000502:physiology*; D059748:Proteolysis / Q000187:drug effects*; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
chemical_list: D015853:Cysteine Proteinase Inhibitors; D007976:Leupeptins; D015842:Serine Proteinase Inhibitors; D014108:Tosylphenylalanyl Chloromethyl Ketone; D007930:Leucine; C108192:aloxistatin; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde