View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by University of Oregon Scholars' Bank THE COMPLETE DEVEWPMENT OF THE DEEP-SEA CIDAROID URCHIN CIDARIS BLAKEI (AGASSIZ, 1878) WITH AN EMPHASIS ON THE HYALINE LAYER by KATHLEEN BENNETT A THESIS Presented to the Department ofBiology and the Graduate School ofthe University ofOregon in partial fulfillment ofthe requirements for the degree of Master ofScience December 2009 11 "The Complete Development ofthe Deep-Sea Cidaroid Urchin Cidaris blakei (Agassiz, 1878) With an Emphasis on the Hyaline Layer," a thesis prepared by Kathleen Bennett in partial fulfillment ofthe requirements for the Master ofScience degree in the Department ofBiology. This thesis has been approved and accepted by: Al 'Sb1ilAld;, C air ofthe Examining Committee Date Committee in Charge: Alan Shanks, Chair Richard Emlet Craig Young Accepted by: Dean ofthe Graduate School III © 2009 Kathleen Bennett IV An Abstract ofthe Thesis of Kathleen Bennett for the degree of Master ofScience in the Department ofBiology to be taken December 2009 Title: THE COMPLETE DEVELOPMENT OF THE DEEP-SEA CIDAROID URCHIN CIDARIS BLAKEI(AGASSIZ, 1878) WITH AN EMPHASIS ON THE HYALINE LAYER Approved: Alan Shanks Living echinoids comprise two major sister clades, the Euechinoidea and the Cidaroidea. Cidaroids first appeared during the lower Permian (~255 mya) and are considered to represent the primitive form ofall other living echinoids. The present study ofCidaris blakei, a deep-sea planktotrophic cidaroid urchin, provides a description ofdevelopment from fertilization through early juvenile stages and is the first report ofa deep-sea organism reared through metamorphosis. Cidaris blakei resembles other cidaroids in its lack ofa cohesive hyaline layer, the absence ofan amniotic invagination for juvenile rudiment formation, and the presence ofa single spine morphotype at metamorphosis. Cidaris blakei differs from other cidaroids in the presence ofan apical v tuft, the extent offenestration ofpostoral skeletal rods, the shape ofjuvenile spines and an extended (14 day) lecithotrophic stage prior to development ofa complete gut. This study includes my co-authored materials. VI CURRICULUM VITAE NAME OF AUTHOR: Kathleen Bennett PLACE OF BIRTH: Cottage Grove, Oregon DATE OF BIRTH: November 8, 1977 GRADUATE AND UNDERGRADUATE SCHOOLS ATTENDED: University ofOregon, Eugene, OR University ofMassachusetts, Amherst MA Southern Oregon University, Ashland, OR DEGREES AWARDED: Master ofScience, Biology, December 2009, University ofOregon Bachelor ofArts, Journalism, March 2002, University ofOregon AREAS OF SPECIAL INTEREST: Marine Biology Comparative Embryology PROFESSIONAL EXPERIENCE: National Science Foundation GK-12 Fellow, University ofOregon, 2007-2009 Graduate Teaching Fellow, University ofOregon, 2007 Vll GRANTS, AWARDS AND HONORS: Best Student Paper, Taking Inquiry into the Field: Curriculum Development/or Elementary Marine Science: Examples from the GK12 "Learning About Where We Live" Project, American Society ofLimnology and Oceanography Ocean Sciences Meetings, 2008. National Science Foundation, GK-12 Graduate Teaching Fellowship, 2007-2009 PUBLICATIONS; Bennett, K. C. and L. Hiebert. 2009. Growing up in the ocean: complex life cycles ofcommon marine invertebrates. Sci. Activ. 46: 18-25. viii ACKNOWLEDGMENTS I am very grateful to my advisor, Alan Shanks, who was supportive ofmy research through all ofits many incarnations. I am also very grateful to the other members ofmy committee, Craig Young and Richard Emlet, for their expert advice during my research and also during the preparation ofthis manuscript. I am also beyond grateful to them for offering the deep-sea course as part oftheir grant and for providing me with one ofthe greatest adventures ofmy life. The faculty at OIMB was very generous with their time and knowledge and some ofmy best education came indirectly, from wandering into their offices and talking science. The microscope where I did most ofmy work was right outside Richard Emlers office and I must give an extra thanks to him for answering the (perhaps hundreds of) questions I shouted through his open doorway. Thanks also to the Maslakova lab, the work I did for Chapter III using monoclonal antibody stains was made possible by the patient counsel of George von Dassow, Svetlana Maslakova and Laurel Hiebert. Thanks to the incredible community ofgraduate students at OIMB who always challenged me intellectually while also challenging me physically--be it braving the sea lions at North Cove while crawling along a transect line in the winter's darkness, swimming through monstrous waves to Saddle Rock, or being part ofa roller derby team made up mostly ofOIMB grad students. Barb Butler was instrumental in the completion of my degree, both in her capacity as a librarian and as a friend. Shirley Pedro also provided a lot ofsupport in the form ofcozy talks, sugary treats and logistical know-how. IX Drs. David Adelson and David McClay mailed me ample amounts ofthe monoclonal antibody McA Tg-HYL that I used in the research described in Chapter III and were also very kind and thorough in answering questions about their protocol. I'm very grateful to the NSF GK-12 grant 0638731, not only for the funding it provided, but also for the additional education. Because ofthis program, I felt much more connected to the community surrounding OIMB and my research felt more relevant and exciting as I shared it with my students and the teachers I worked with. Thanks to Jan Hodder, Alan Shanks and Trish Mace for bringing the program to OIMB and for coordinating it. Finally, thanks to my family who provided me with a lot ofsupport and to Cameron Bishop, who finds science as exciting as I do and who was always willing to discuss ideas and make midnight microscope missions to the lab. x This thesis is dedicated to my parents, who instilled a love oflearning in me at an early age and who have always make me feel that I am capable ofanything. Xl TABLE OF CONTENTS Chapter Page 1. GENERAL mTRODUCTION 1 II. LARVAL DEVELOPMENT AND METAMORPHOSIS OF THE DEEP-SEA CIDAROID URCHm CIDARIS BLAKEI................................................................... 6 Introduction............................................................................................................ 6 Materials and Methods........................................................................................... 9 Results.................................................................................................................... 12 Discussion............................................ 21 III. THE CIDAROID HYALmE LAYER DIFFERS m FORM AND FUNCTION FROM THE EUECHmOID HYALmE LAYER 34 Introduction..................................... 34 Materials and Methods........................................................................................... 36 Results.................................................................................................................... 39 Discussion 46 Conclusion 49 xii Chapter Page IV. CONCLUDING SUMMARY 51 BffiLIOGRAPHY 55 XIIl LIST OF FIGURES Figure Page Chapter I 1. Two images showing Cidaris blakei adults.......... 2 Chapter II 1. Light micrographs of C. blakei from immediately after fertilization to nine days after fertilization............ 13 2. Scanning Electron Micrographs ofa 4 day-old C. blakei gastrula showing ectodermal pits. 15 3. Light micrographs of C. blakei plutei from 10 days to 79 days after fertilization................................................................................................................... 16 4. Light micrographs ofadvanced larvae ofC. blakei with juvenile structures evident on the larval body.. 17 5. Light micrographs ofa competent larva and juveniles ofC. blakei. 19 6. Light micrographs ofC. blakei larvae from different temperature treatments. Plutei were maintained in scintillation vials and moved up a thermal gradient, from 11 °C to 22 °C, over a four day period. As a control, plutei were kept in scintillation vials for four days at 11 °C. 20 7. Two micrographs showing the difference between the juvenile spine shape of Eucidaris thouarsi and Cidaris blakei 31 XIV Figure Page Chapter III 1. Light micrographs ofmulti-cell stages ofcidaroid sea urchins Cidaris blakei and Eucidaris tribuloides showing a thin extracellular layer surrounding the blastomeres 40 2. Confocal projections offertilized eggs ofStrongylocentrotus purpuratus labeled with phalloidin (orange) and the hyalin-specific monoclonal antibody McA Tg-HYL 41 3. Confocal projections ofa fertilized egg ofEucidaris tribuloides labeled with phalloidin (orange). This fertilized egg was also treated with the monoclonal antibody McA Tg-HYL, but it did not react................................................................ 42 4. Confocal projections ofmulti-cell stages ofthree species ofsea urchin labeled with phalloidin and the hyalin-specific monoclonal antibody McA Tg-HYL 43 5. Confocal projection ofa fertilized egg ofS. purpuratus that has been fixed in the same 10% buffered formalin solution as C. blakei showing a hyaline layer clearly labeled with McA Tg-HYL.............................................................................. 44 6. Confocal projections ofblastula-stage embryos labeled with phalloidin (orange) and the hyalin-specific monoclonal antibody McA Tg-HYL (blue) 45 7. Confocal z-projection ofa two-cell embryo ofa clypeasteroid, Dendraster excentricus, labeled with phalloidin