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Neutral endopeptidase-24.11 (NEP) deactivates PDGF- and TGF-beta-induced cell shape changes in invertebrate
E Caselgrandi1, D Kletsas, E Ottaviani
1Department of Biomedical Sciences, Hygiene and Microbiology Section, University of Modena and Reggio Emilia, Modena, 41100, Italy.
Insights
Mussel immunocytes express CD10 (neutral endopeptidase-24.11), an enzyme that interacts with growth factors like PDGF-AB and TGF-beta1. This enzyme influences cell shape changes, suggesting a conserved biological pathway across species.
Area of Science:
- Immunology
- Marine Biology
- Biochemistry
Background:
- Mollusc immunocytes play a crucial role in invertebrate immunity.
- CD10, also known as neutral endopeptidase-24.11 (NEP), is a cell surface enzyme with known roles in vertebrate systems.
- Growth factors such as PDGF-AB and TGF-beta1 are key regulators of cellular processes.
Purpose of the Study:
- To investigate the expression and function of CD10/NEP in Mytilus galloprovincialis immunocytes.
- To explore the interaction between NEP and growth factors PDGF-AB and TGF-beta1 in these cells.
- To determine if NEP influences growth factor-induced cellular responses.
Main Methods:
- Cytofluorimetric assay to detect CD10 expression on immunocytes.
- Spectrofluorimetric analysis to measure NEP-like activity.
- Computer-assisted microscopic image analysis for cell shape changes.
- Western blot analysis to assess protein cleavage.
Main Results:
- Mytilus immunocytes express CD10, identical to NEP.
- Growth factors PDGF-AB and TGF-beta1 increased NEP-like activity in membrane preparations.
- NEP deactivated PDGF-AB and TGF-beta1-induced immunocyte shape changes, but without direct proteolysis.
Conclusions:
- A functional interplay exists between growth factors and NEP in invertebrate immunocytes.
- This interaction mirrors previously observed mechanisms in vertebrate cells.
- NEP's role in modulating growth factor signaling in Mytilus suggests conserved biological functions.
Abstract:
Using a cytofluorimetric assay, we found that immunocytes of the mollusc, Mytilus galloprovincialis, express CD10, a surface antigen known to be identical to neutral endopeptidase-24.11 (NEP). The spectrofluorimetric analysis demonstrates that the growth factors PDGF-AB and TGF-beta1 provoke an increase in NEP-like activity in membrane preparations from the immunocytes, but have no effect on the soluble form in the serum. On the other hand, computer-assisted microscopic image analysis reveals that NEP deactivates the PDGF-AB- and TGF-beta1-induced shape changes in immunocytes. However, Western blots show that, in solution, NEP does not cleave PDGF-AB or TGF-beta1, indicating that the inactivation is not due to proteolysis. These results suggest a functional interplay in invertebrate immunocytes between growth factors and NEP, as previously shown in vertebrate cells.
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