title: Cardosins improve neuronal regeneration after cell disruption: a comparative expression study.
abstract: The establishment of primary cell cultures is invaluable for studying cell and molecular biological questions. Although primary cell cultures more closely resemble and function like in the native environment, during the culture establishment the cells undergo several changes including the damage sustained during their removal from original tissue. The resultant cells have to rebalance the expression of their processing molecules to ascertain matrix signalling that ensure cell adaptation and consequent proliferation. Hence, we used cardosin, a novel plant enzyme for tissue disaggregation, for isolating and culturing neuronal cells from embryonic rats. The present investigation reports the molecular events, mainly related with matrix metalloproteinases (MMPs)/tissue inhibitor of metalloproteinase (TIMPs) expression, which could substantiate the superior neurite outgrowth and dendritic extension previously described. It was observed that 24 h after primary culture establishment, MMP-2 and MMP-9 messenger RNA (mRNA) are significantly upregulated, while the expression of TIMP-1 and TIMP-2 is unaltered. Regarding the role of laminin in neuronal pathfinding, it was found that the use of anti-laminin antibody and arginine-glycine-aspartate (RGD) peptide exerted inhibitory effects on neurite outgrowth after mechanical lesion where the expression of MMP-9 and TIMP-1 is upregulated under non-permissive conditions in response to mechanical injury.
authors.affiliation: Centre for Environmental and Marine Studies, Department of Biology, University of Aveiro, Aveiro, Portugal. asduarte@ua.pt
authors.forename: Ana Sofia
authors.initials: AS
authors.lastname: Duarte
authors.forename: Emília P
authors.initials: EP
authors.lastname: Duarte
authors.forename: António
authors.initials: A
authors.lastname: Correia
authors.forename: Euclides
authors.initials: E
authors.lastname: Pires
authors.forename: Marlene T
authors.initials: MT
authors.lastname: Barros
mesh_terms: D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000494:pharmacology*; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000187:drug effects / Q000502:physiology; D035264:Flowers / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects; D007797:Laminin / Q000502:physiology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D009416:Nerve Regeneration / Q000187:drug effects*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D010940:Plant Proteins / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D007797:Laminin; D010940:Plant Proteins; D012333:RNA, Messenger; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; D016282:Aspartic Acid Endopeptidases; C097455:aspartic proteinases, Cynara cardunculus; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9