A Functional Study of Transforming Growth Factor-Beta from the Gonad of Pacific Oyster Crassostrea gigas
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The transforming growth factor (TGF)-β superfamily is a group of important growth factors involved in multiple processes such as differentiation, cell proliferation, apoptosis and cellular growth. In the Pacific oyster Crassostrea gigas, the oyster gonadal (og) TGF-β gene was recently characterized through genome-wide expression profiling of oyster lines selected to be resistant or susceptible to summer mortality. Og TGF-β appeared specifically expressed in the gonad to reach a maximum when gonads are fully mature, which singularly contrasts with the pleiotropic roles commonly ascribed to most TGF-β family members. The function of og TGF-β protein in oysters is unknown, and defining its role remains challenging. In this study, we develop a rapid bacterial production system to obtain recombinant og TGF-β protein, and we demonstrate that og TGF-β is processed by furin to a mature form of the protein. This mature form can be detected in vivo in the gonad. Functional inhibition of mature og TGF-β in the gonad was conducted by inactivation of the protein using injection of antibodies. We show that inhibition of og TGF-β function tends to reduce gonadic area. We conclude that mature og TGF-β probably functions as an activator of germ cells development in oyster.
KeywordsTransforming growth factor-β Bacterial expression In vivo antibody inhibition Reproduction Crassostrea gigas Marine bivalve
The present research project was supported by “Europole Mer” (www.europolemer.eu; project “OxyGenes”) and by ANR (project “Gametogenes” ANR-08-GENM-041). The authors are grateful for the financial support from the ‘Marine Genomics Europe’ network of excellence (European Commission contract number GOCE-CT-2004-505403). E. Fleury was funded by Ifremer and a Région Basse-Normandie doctoral grant. We thank all the staff of the Argenton experimental hatchery for conditioning oysters. The authors are indebted to J.Y. Daniel, V. Quilien and C. Quéré for advice and technical assistance. We thank J. Moal for her comments on the manuscript and H. McCombie for editing the English language.
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