Recent studies demonstrated that the ribonuclease RNASET2 modulates inflammatory processes in both vertebrates and invertebrates. This protein chemoattracts macrophages in vivo and its expression significantly increases after bacterial infections. Moreover recent data obtained in our laboratory, demonstrated that the injection of the human recombinant protein RNASET2 in the leech body wall, a consolidated invertebrate model for studying both immune response and tissue regeneration, induces not only macrophages recruitment, but also a massive connective tissue remodeling. Based on these data, here we evaluated a possible direct or indirect role of the leech rHvRNASET2, recently cloned in our laboratory in the synthesis of new collagen. The connective tissue reorganization and the cell types involved in this process were characterized in rHvRNASET2 injected leeches by means of histochemical staining and morphological analyses at optical and electron microscopes. The expression of newly synthesized Pro-collagen1α1, together with that of fibroblast receptors (FGFb), was localized at tissue level by immunofluorescence. Moreover, to evaluate the expression profile of Pro-collagen1α1, western blot experiments were performed on protein extracts from leech body wall retrieved after different time elapse from rHvRNASET2 injection. Double immunofluorescence assays were performed to correlate fibroblasts activation, collagen fibrils production and the consequent connective tissue remodeling with the expression of HvRNASET2. The data reported in this work provide compelling evidence in support of a pleiotropic role for RNASET2 in orchestrating an evolutionarily conserved cross-talk between inflammatory response and regenerative process, based on macrophages recruitment and fibroblasts activation coupled to a massive extracellular remodeling.
The recombinant HvRNASET2 protein induces a connective tissue remodeling in the invertebrate model medicinal leech Hirudo verbana
N Baranzini;F Acquati;G Tettamanti;A Grimaldi
2019-01-01
Abstract
Recent studies demonstrated that the ribonuclease RNASET2 modulates inflammatory processes in both vertebrates and invertebrates. This protein chemoattracts macrophages in vivo and its expression significantly increases after bacterial infections. Moreover recent data obtained in our laboratory, demonstrated that the injection of the human recombinant protein RNASET2 in the leech body wall, a consolidated invertebrate model for studying both immune response and tissue regeneration, induces not only macrophages recruitment, but also a massive connective tissue remodeling. Based on these data, here we evaluated a possible direct or indirect role of the leech rHvRNASET2, recently cloned in our laboratory in the synthesis of new collagen. The connective tissue reorganization and the cell types involved in this process were characterized in rHvRNASET2 injected leeches by means of histochemical staining and morphological analyses at optical and electron microscopes. The expression of newly synthesized Pro-collagen1α1, together with that of fibroblast receptors (FGFb), was localized at tissue level by immunofluorescence. Moreover, to evaluate the expression profile of Pro-collagen1α1, western blot experiments were performed on protein extracts from leech body wall retrieved after different time elapse from rHvRNASET2 injection. Double immunofluorescence assays were performed to correlate fibroblasts activation, collagen fibrils production and the consequent connective tissue remodeling with the expression of HvRNASET2. The data reported in this work provide compelling evidence in support of a pleiotropic role for RNASET2 in orchestrating an evolutionarily conserved cross-talk between inflammatory response and regenerative process, based on macrophages recruitment and fibroblasts activation coupled to a massive extracellular remodeling.
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