Multiple vitellogenins and product yolk proteins in striped bass, Morone saxatilis: molecular characterization and processing during oocyte growth and maturation V. N. Williams • B. J. Reading • N. Hiramatsu • H. Amano • N. Glassbrook • A. Hara • C. V. Sullivan Received: 23 March 2013 / Accepted: 19 August 2013 / Published online: 5 September 2013 Ó Springer Science+Business Media Dordrecht 2013 Abstract The multiple vitellogenin (Vtg) system of striped bass, a perciform species spawning nearly neutrally buoyant eggs in freshwater, was investi- gated. Vitellogenin cDNA cloning, Western blotting of yolk proteins (YPs) using Vtg and YP type-specific antisera, and tandem mass spectrometry (MS/MS) of the YPs revealed the complex mechanisms of yolk formation and maturation in this species. It was discovered that striped bass possesses a tripartite Vtg system (VtgAa, VtgAb, and VtgC) in which all three forms of Vtg make a substantial contribution to the yolk. The production of Vtg-derived YPs is generally similar to that described for other perciforms. How- ever, novel amino-terminal labeling of oocyte YPs prior to MS/MS identified multiple alternative sites for cleavage of these proteins from their parent Vtg, revealing a YP mixture far more complex than reported previously. This approach also revealed that the major YP product of each form of striped bass Vtg, lipovitellin heavy chain (LvH), undergoes limited degradation to smaller polypeptides during oocyte maturation, unlike the case in marine fishes spawning buoyant eggs in which LvHAa undergoes extensive proteolysis to osmotically active free amino acids. These differences likely reflect the lesser need for hydration of pelagic eggs spawned in freshwater. The detailed characterization of Vtgs and their proteolytic fate(s) during oocyte growth and maturation estab- lishes striped bass as a freshwater model for investi- gating teleost multiple Vtg systems. Keywords Striped bass Á Vitellogenin Á Yolk Á Post-vitellogenic oocytes Á Ovulated eggs Á Maturation Á Mass spectrometry Introduction Vitellogenins (Vtgs) are glycolipophosphoprotein precursors to yolk in egg-laying vertebrates that are produced by the liver and released into the Electronic supplementary material The online version of this article (doi:10.1007/s10695-013-9852-0) contains supple- mentary material, which is available to authorized users. V. N. Williams (&) Á B. J. Reading Á C. V. Sullivan Department of Biology, College of Agriculture and Life Sciences, North Carolina State University, 127 David Clark Labs, Raleigh, NC 27695, USA e-mail: vnwillia@ncsu.edu; vnwillia@gmail.com N. Hiramatsu Á A. Hara Graduate School of Fisheries Sciences, Hokkaido University, 3-1-1, Minato, Hakodate, Hokkaido 041-8611, Japan H. Amano School of Marine Biosciences, Kitasato University, 1-15-1 Kitasato, Minami, Sagamihara, Kanagawa 252-0373, Japan N. Glassbrook Genomics Sciences Laboratory, College of Agriculture and Life Sciences, North Carolina State University, Raleigh, NC 27695, USA 123 Fish Physiol Biochem (2014) 40:395–415 DOI 10.1007/s10695-013-9852-0