Poccia Research Lab

Research Description

• During maturation of most animal sperm cells, sperm-specific nuclear proteins replace somatic histones, DNA synthesis, RNA synthesis, and cell division cease, and the chromatin becomes highly compacted in preparation for the sperm cell's journey to the egg. After fertilization, the dormant sperm nucleus must become activated in order to participate in normal cellular events such as replication, transcription, and mitosis. The deactivation of the male germ cell nucleus during spermatogenesis and its reactivation following fertilization can serve as model systems for the study of gene activity, replication, chromatin and nuclear envelope assembly, and higher order chromatin structure.
• We use in vitro cultures of sea urchin testis fragments, polyspermically fertilized eggs, microinjection, and cell-free systems to study at the biochemical level transitions in nuclear composition, architecture, and activity during spermatogenesis and fertilization. Using techniques such as gel electrophoresis, in situ hybridization, fluorescence microscopy, flow cytometry, organ culture and subcellular fractionation, we are attempting to understand the molecular basis of sperm nuclear differentiation and male pronuclear formation.
• We have been able to reconstruct many of the pronuclear transformations in a test tube thus making possible much more detailed biochemical investigations.Most recently, we have focused on formation of the male pronuclear envelope emphasizing the role of proteins and lipids in membrane fusion events.
• IMAGES
  
  
  • Click here to view an enlarged (though inverted) sea urchin: sea urchin.
  • This is a section of a sea urchin testis reacted with a radioactive RNA probe that reveals sites of expression of the Sp H2B gene which codes for a testis-specific nuclear protein (histone) during spermatogenesis. The silver grains correspond to expression of its mRNA and are localized at the periphery of the tissue where the spermatogonia are dividing.
  • This is a section of a sea urchin embryo at the pluteus larva stage. The nuclei of the cells are stained blue with a Hoechst fluorescent dye and unknown molecules of the extracellular matrix are stained green with a monoclonal antibody isolated in our lab.
  • Male pronucleus at 15 minutes post-fertilization showing DNA (blue) and endoplasmic reticulum derived nuclear envelope (red). Confocal image from egg microinjected with lipophilic dye.
  • An unfertilized sea urchin egg with endoplasmic reticulum labelled in red and plasma membrane in green. Close apposition of tubules of ER with the plasma membrane.

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Professional Activities

I am an adjunct professor in the Molecular and Cellular Biology Program at the University of Massachusetts, Amherst and consultant for Hematech, LLC.

I serve as editor of the Reproductive Biology Section of the Journal of Experimental Zoology, an associate editor for Molecular Reproduction and Development and am a member of the Editorial Board of The Biological Bulletin of the Scientific Advisory Board of Signal Transduction.

Publications

• Domart, M.C., Hobday, T.M.C., Peddie, C.J., Chung, G.H.C., Wang, A., Yeh, K., Jethwa, N., Zhang, Q., Wakelam, M.J.O.,  Woscholski, R., Byrne, R.D., Collinson, L.M., Poccia, D.L. and Larijani, B. (2012). The function of diacylglycerol in controlling ER morphology and nuclear envelope fusion from echinoderms to mammals, PLoS ONE 7: e51150. doi:10.1371
• Larijani, B. and Poccia, D. (2012). Effects of Phosphoinositides and their Derivatives on Membrane Morphology and Function,  Curr.Topics Microbiol Immunol. 362: 99-110 In, Phosphoinositides and Disease,. Chap 5, M. Falasca, ed., Springer.
  
• Byrne RD, Applebee C, Poccia DL, Larijani B (2012) Dynamics of PLCγ and Src Family Kinase 1 Interactions during Nuclear Envelope Formation Revealed by FRET-FLIM. PLoS ONE 7: e40669. doi:10.1371.
• Chatzidimitriadou,  Z. and Poccia, D. (2010). Localization of PLCg and Nuclear Envelopes in Telophase of Sea Urchin Embryos. Molec. Reprod. Devel. 77: 737.
• Dumas, F., Byrne, R.D., Vincent, B., Hobday, T.M.C., Poccia, D. and Larijani, B. (2010) Spatial Regulation of Membrane Fusion Controlled by Modification of Phosphoinositides, PLOS ONE 5, e12208 doi:10.1371.
• Byrne, R. Poccia, D. and Larijani, B. (2009). The Role of Phospholipase C in Nuclear Envelope Assembly.  Clinical Lipidology 4: 103-112.
• Byrne, R. Larijani, B. and Poccia, D. (2009). Tyrosine Kinase Regulation of Nuclear Envelope Assembly.  Adv. Enzyme Regul. 49: 148-156.
• Larijani, B. and Poccia, D. (2009). Nuclear Envelope Formation: Mind the Gaps.  Annu. Rev. Biophysics 38:107-124.
• Poccia, D. and Larijani, B. (2009). Phosphatidylinositol Metabolism and Membrane Fusion. Biochem J. 418: 233-246.
• Garnier-Lhomme, M., Byrne, R.D., Hobday, T.M.C., Gschmeissner, S., Woscholski, R., Poccia, D., Dufourc, E.J. and Larijani, B. (2009) Nuclear Envelope Remnants: Fluid Membranes Enriched in Sterols and Polyphosphoinositides, PLoS One, 4: e4255.
• Garnier-Lhomme, M., Dufourc, E.J., Larijani, B., and Poccia, D.(2009). Lipid Quantification and Structure Determination of  Nuclear Envelope Precursor Membranes in the Sea Urchin  In, Lipid Signaling Protocols, Methods in Molecular Biology, B. Larijani, R. Wolchoski and C. Rosser, eds. Humana Press, Chap. 6, pp. 89-110.
• Byrne, R., Zhandre, V., Larijani, B. and Poccia, D. (2008). Nuclear Envelope Formation In Vitro: A Sea Urchin Egg Cell-Free System. In, The Nucleus: Physical Properties and Imaging Methods, Principles and Protocols, Methods in Molecular Biology, Vol 2. R. Hancock, ed., Humana Press, Chap. 12, pp. 205-220.
• Larijani, B. and Poccia, D. (2007). Protein and Lipid Signaling in Membrane Fusion: Nuclear Envelope Assembly.  Signal Transduction 7: 142-153.
• Poccia, D. (ed.) (2007). The Sea Urchin as a Model System for Investigation of Signaling Mechanisms.  Signal Transduction, 7: 99-101.
• Byrne, R., Garnier-Lhomme, M., Han, K., Dowicki, M., Michael, N., Totty, N., Zhendre, V., Cho, A., Pettitt, T.R.,Wakelam, M.J.O., Poccia, D and Larijani, B. (2007). PLCg is Enriched on Poly-Phosphoinositide-Rich Vesicles to Control Nuclear Envelope Assembly, Cellular Signalling 19: 913-922.
• Poccia, D. (2006). Editorial: Beyond the Model Organism. J. Exp. Zool. 305A: 537.
• Barona, T., Larijani, B. and Poccia, D. (2005).  Dynamics of Nuclear Envelope Formation. Endocrinologia, Metabolismo & Nutrição 14: 107-114.
• Barona, T., Byrne, R., Pettit, T.R., Wakelam, M.J.O., Larijani, B. and Poccia, D. (2005). Diacylglycerol induces fusion of nuclear envelope membrane precursor vesicles. J. Biol. Chem. 280: 41171-41177.
• Byrne, R., Barona, T., Garnier, M., Koster, G., Katan, M., Poccia, D. and Larijani, B. (2005).Nuclear Envelope Assembly is Promoted by Phosphoinositide-Specific PLC with Selective Recruitment of Phosphatidylinositol Enriched Membranes. Biochem. J . 387: 393-400.
• Stephens, S. Beyer, B. Balthazar-Stablein, U., Duncan, R., Kostacos, M., Lukoma, M., Green, G.R. and Poccia, D. (2002). Two Kinase Activities are Sufficient for Sea Urchin Sperm Chromatin Decondensation In Vitro, Molec. Reprod. Devel. 62: 496-503.
• Poccia, D., Barona, T., Collas, P. and Larijani, B. (2002). Nuclear Envelope Assembly in Gametes and Pronuclei. In, Nuclear Envelope Assembly in Embryos and Somatic Cells. (P. Collas, ed.) Klewer?Plenum, New York, pp. 111-130.
• Green G.R. , Ferlitta R.R., Walkenhorst W.F. and Poccia D.L. (2001). Linker DNA Destabilizes Condensed Chromatin Structure. Biochem. Cell Biol. 79: 349-363.
• Larijani, B., Barona, T. and Poccia, D. (2001). Role for Phosphatidylinositol in Nuclear Envelope Formation. Biochem. J. 356: 495-501.
• Larijani, B., Poccia, D.L. and Dickinson, L.C. (2000). Phospholipid Identification and Quantification of Membrane Vesicle Subfractions by Two-dimensional 31P-1H NMR Spectroscopy with Isotropic Protein Mixing, Lipids 35: 1289-97.
• Collas, P., Barona, T. and Poccia, D. (2000). Rearrangements of Sea Urchin Egg Cytoplasmic Membrane Domains at Fertilization. Eur. J. Cell Biol. 79: 10-16.
• Collas, P. and Poccia, D. (2000). Membrane Fusion Events during Nuclear Envelope Assembly. Subcellular Biochem. 34: 273-302.
• Collas, P. and Poccia, D. (1998). Methods for Studying In Vitro Assembly of Male Pronuclei Using Extracts from Marine Invertebrates: Sea Urchins and Surf Clams. Meth. Cell Biol., 53: 417-452.
• Collas, P. and Poccia D. (1998). Remodeling the Sperm Nucleus into a Male Pronucleus at Fertilization. Theriogenology 49: 67-81.
• Poccia, D. and Collas, P. (1997). Nuclear Envelope Dynamics During Male Pronuclear Development, Devel. Growth Differ. 39, 541-550.
• Collas, P., Thompson, L., Fields, A.P., Poccia, D.L. and Courvalin, J.-C. (1997). PKC-Mediated Interphase Lamin B Phosphorylation and Solubilization, J. Biol. Chem.272: 21274-21280.
• Collas, P., Courvalin, J.-C. and Poccia, D.L. (1996). Targeting of Membranes to Sea Urchin Sperm Chromatin is Mediated by an LBR-Like Integral Membrane Protein. J. Cell Biol. 135: 1715-1725.
• Collas, P. and Poccia, D.L. (1996). Conserved Binding Recognition Elements of Sperm Chromatin, Sperm Lipophilic Structures and Nuclear Envelope Precursor Vesicles. Eur J. Cell Biol. 71: 22-32.
• Vafa, O., Goetzl, L., Poccia, D. and Nishioka, D. (1996). Localization and Characterization of Blastocoelic Extracellular Matrix Antigens in Early Sea Urchin Embryos and Evidence for Their Proteolytic Modification during Gastrulation. Differentiation 60: 129-138.
• Collas, P. and Poccia, D. (1996). Distinct Egg Cytoplasmic Membrane Vesicles Differing in Binding and Fusion Properties Contribute to Sea Urchin Male Pronuclear Envelopes Formed In Vitro J. Cell Sci. 109: 1275-1283.
• Poccia. D. and Collas, P. (1996). Transforming Sperm Nuclei into Male Pronuclei In Vivo and In Vitro. Curr. Topics Develop. Biol.34: 25-88.
• Poccia, D. (1995). Male Germ Line Specific Histones of Sea Urchins and Sea Stars. In, Advances in Spermatozoal Taxonomy and Phylogeny, (B.G.M. Jamieson, J. Ausio and J.-L. Justine, eds.), Mémoires du Muséum National d'Histoire Naturelle (Paris), pp. 475-490.
• Green, G.R., Collas, P., Burrell, A. and Poccia, D. (1995). Histone Phosphorylation during Sea Urchin Development. Seminars in Cell Biology 6: 219-227.
• Collas, P., Pinto-Correia, C. and Poccia, D. (1995). Lamin Dynamics during Sea Urchin Male Pronucleus Formation In Vitro Exp. Cell Res. 219: 687-698.
• Collas, P. and Poccia, D. (1995). Formation of the Sea Urchin Male Pronucleus In Vitro: Membrane-Independent Chromatin Decondensation and Nuclear Envelope- Dependent Nuclear Swelling. Molec. Reprod. Devel. 42:, 106-113.
• Collas, P. and Poccia, D.L. (1995). Lipophilic Organizing Structures of Sperm Nuclei Target Membrane Vesicle Binding And Are Incorporated into the Nuclear Envelope. Devel. Biol. 169: 123-135.
• Pinto-Correia, C., Poccia, D.L., Chang,T. and Robl, J.M. (1994). Dephosphorylation of Sperm Midpiece Antigens Initiates Aster Formation in Rabbit Oocytes. Proc. Natl. Acad. Sci. USA. 91: 7894-7898.
• Poccia, D.L. (1994). Molecular Aspects of Spermatogenesis, R.G. Landes, Austin.
• Cameron, L.A. and Poccia, D.L. (1994). In Vitro Development of the Sea Urchin Male Pronucleus. Devel. Biol. 162: 568- 578.
• Green, G.R., Balhorn, R., Poccia, D.L. and Hecht, N.B. (1994). Synthesis and Processing of Mammalian Protamines and Transition Proteins. Molec. Reprod. Develop. 37: 255-263.
• Green, G.R., Lee, H.-J. and Poccia, D.L. (1993). Phosphorylation Weakens DNA Binding by Peptides Containing Multiple "SPKK" Sequences. J. Biol. Chem. 268:11247-11255.
• Cothren, C.C. and Poccia, D.L. (1993). Two Steps Required for Male Pronucleus Formation in the Sea Urchin Egg. Exp. Cell Res. 205:126-133.
• Poccia, D.L. and Green, G.R. (1992). Packaging and Unpackaging the Sea Urchin Sperm Genome. Trends in Biochem. Sci. 17:223-227.
• Poccia, D. (1991).  Sp Histones and Chromatin Structure in Male Germ Line Nuclei and Male Pronuclei of the Sea Urchin.  In, Comparative Spermatology: 20 Years After.  (B. Baccetti, ed.), Raven Press, NY, pp. 61-66.
• Poccia, D.L. (1991).  George Emile Palade.  In, The Nobel Prize Winners: Physiology or Medicine.  (F.N. Magill, ed.), Salem Press, Pasadena, pp. 1190-1199.
• Poccia, D.L. (1991).  Christian De Duve.  In, The Nobel Prize Winners: Physiology or Medicine.  (F.N. Magill, ed.), Salem Press, Pasadena, pp. 1178-1187.
• Green, G.R., Patel, J.C., Hecht, N.B. and Poccia, D.L. (1991).  A Complex Pattern of H2A Phosphorylation in the Mouse Testis.  Exp. Cell Res. 195:8-12.
• Poccia, D.L. (1991).  In Situ Hybridization.  In, Magill's Survey of Science: Life Sciences.  (F.N. Magill, ed.), Salem Press, Pasadena, pp. 1408-1413.
• Poccia, D., Pavan, W. and Green, G.R. (1990).  6DMAP Inhibits Chromatin Decondensation but Not Sperm Histone Kinase in Sea Urchin Male Pronuclei.  Exp. Cell Res. 188:226-234.
• Green, G.R., Gustavson, L. and Poccia, D. (1990).  Phosphorylation of Plant H2A Histones.  Plant Physiol. 93:1241-1245.
• Vodicka, M., Green, G.R. and Poccia, D.L. (1990).  Sperm Histones and Chromatin Structure of the "Primitive" Sea Urchin Eucidaris tribuloides.  J. Exp. Zool. 256:179-188.
• Nishioka, D., Ward, R.D., Poccia, D., Kostacos, C. and Minor, J.E. (1990).  Localization of Bindin Expression During Sea Urchin Spermatogenesis.  Molec. Reprod. Devel. 27:181-190.
• Green, G.R. and Poccia, D.L. (1989).  Phosphorylation of Sea Urchin Histone CS H2A.  Devel. Biol. 134:413-419.
• Poccia, D., Lieber, T. and Childs, G. (1989).  Histone Gene Expression During Sea Urchin Spermatogenesis: An in situ Hybridization Study.  Molec. Reprod. Devel. 1:219-229.
• Poccia, D. (1989).  Reactivation and Remodeling of the Sperm Nucleus Following Fertilization.  In, The Molecular Biology of Fertilization.  (H. Schatten and G. Schatten, eds.), Acad. Press, NY, pp. 115-135.
• Poccia, D. (1988).  In Vitro Differentiation of Male Germ Line Cells from Sea Urchin Testis.  J. Exp. Zool. 246:57-64.
• Green, G.R. and Poccia, D. (1988).  Interaction of Sperm Histone Variants and Linker DNA During Spermiogenesis in the Sea Urchin.  Biochemistry 27:619-625.
• Nishioka, D., Trimmer, J.S., Poccia, D. and Vacquier, V.D. (1987).  Changing Localizations of Site-Specific Surface Antigens During Sea Urchin Spermiogenesis.  Exp. Cell Res. 173:606-617.
• Simpson, M.V. and Poccia, D. (1987).  Sea Urchin Testicular Cells Evaluated by Fluorescence Microscopy of Unfixed Tissue.  Gamete Res. 17:131-144.
• Poccia, D.L., Simpson, M.V. and Green, G.R. (1987).  Transitions in Histone Variants During Sea Urchin Spermatogenesis.  Devel. Biol. 121:445-453.
• Poccia, D. (1987).  Regulation of Chromatin Condensation and Decondensation in Sea Urchin Pronuclei.  In, Molecular Regulation of Nuclear Events in Mitosis and Meiosis.  (R.A. Schlegel, M.S. Halleck and P.N. Rao, eds.), Acad. Press, NY, Chap. 6, pp. 149-177.
• Poccia, D. (1986).  Remodeling of Nucleoproteins During Gametogenesis, Fertilization, and Early Development.  Inter. Rev. Cytol. 105:1-65.
• Poccia, D.L. and Green, G.R. (1986).  Nuclei and Chromosomal Proteins.  Meth. Cell Biol. 27, Chap 9, pp.153-173.
• Poccia, D., Wolff, R., Kragh, S. and Williamson, P. (1985).  RNA Synthesis in Male Pronuclei of the Sea Urchin.  Biochim. Biophys. Acta 824:349-356.
• Green, G.R. and Poccia, D.L. (1985).  Phosphorylation of Sea Urchin Sperm H1 and H2B Histones Precedes Chromatin Decondensation and H1 Exchange During Pronuclear Formation.  Devel. Biol. 108:235-245.
• Poccia, D., Greenough, T., Green, G.R., Nash, E., Erickson, J. and Gibbs, M. (1984).  Remodeling of Sperm Chromatin Following Fertilization: Nucleosome Repeat Length and Histone Variant Transitions in the Absence of DNA Synthesis.  Devel. Biol. 104:274-286.
• Poccia, D.L. (1982).  Biochemical Aspects of Sperm Nucleus Activation by Egg Cytoplasm.  J. Wash. Acad. Sci. 72: 24-34.
• Green, G.R., Poccia, D. and Herlands, L. (1982).  A Multisample Device for Electroelution, Concentration, and Dialysis of Proteins from Fixed and Stained Gel Slices.  Anal. Biochem. 123:66-73.
• Herlands, L., Allfrey, V.G. and Poccia, D. (1982).  Translational Regulation of Histone Synthesis in the Sea Urchin, Strongylocentrotus purpuratus.  J. Cell Biol. 94:219-223.
• Salik, J., Herlands, L., Hoffman, H.-P. and Poccia, D. (1981).  Electrophoretic Analysis of the Stored Histone Pool of Unfertilized Sea Urchin Eggs: Quantification and Identification by Antibody Binding.  J. Cell Biol. 90:385-395.
• Krystal, G.W. and Poccia, D.L. (1981).  Phosphorylation of Cleavage Stage Histone H1 in Mitotic and Prematurely Condensed Chromosomes.  Exp. Cell Res. 134:41-48.
• Savic, A., Richman, P., Williamson, P. and Poccia, D. (1981).  Alterations in Chromatin Structure During Early Sea Urchin Embryogenesis.  Proc. Natl. Acad. Sci. USA 78:3706-3710.
• Poccia, D., Salik, J. and Krystal, G. (1981).  Transitions in Histone Variants of the Male Pronucleus Following Fertilization and Evidence for a Maternal Store of Cleavage-Stage Histones in the Sea Urchin Egg.  Devel. Biol. 82:287-296.
• Krystal, G.W. and Poccia, D. (1979).  Control of Chromosome Condensation in the Sea Urchin Egg.  Exp. Cell Res. 123:207-219.
• Poccia, D.L., Palevitz, B.A., Campisi, J. and Lyman, H. (1979).  Fluorescence Staining of Living Cells with Fluorescamine.  Protoplasma 98:91-113.
• Savic, A. and Poccia, D. (1978).  Separation of Histones from Contaminating Ribosomal Proteins by Two-Dimensional Gel Electrophoresis.  Anal. Biochem. 88:573-579.
• Poccia, D., Krystal, G., Nishioka, D. and Salik, J. (1978).  Control of Sperm Chromatin Structure by Egg Cytoplasm in the Sea Urchin.  In, ICN-UCLA Symposium on Molecular and Cellular Biology: Cell Reproduction.  (E.R. Dirksen, D. Prescott and C.F. Fox, eds.), Acad. Press, NY, Chap. 17, pp. 197-206.
• Poccia, D.L., LeVine, D. and Wang, J.C. (1978).  Activity of a DNA Topoisomerase (Nicking-Closing Enzyme) During Sea Urchin Development and the Cell Cycle.  Devel. Biol. 64:273-283.
• Maller, J., Poccia, D., Nishioka, D., Kidd, P., Gerhart, J. and Hartman, H. (1976).  Spindle Formation and Cleavage in Xenopus Eggs Injected with Centriole Containing Fractions from Sperm.  Exp. Cell Res. 99:285-294.
• Poccia, D.L. and Hinegardner, R.T. (1975).  Developmental Changes in Chromatin Proteins of the Sea Urchin from Blastula to Mature Larva.  Devel. Biol. 45:81-89.
• Hill, R.J., Poccia, D.L. and Doty, P. (1971).  Towards a Total Macromolecular Analysis of Sea Urchin Embryo Chromatin.  J. Mol. Biol. 61:445-462.
• Chetsanga, C., Poccia, D.L., Hill, R.J. and Doty, P. (1970).  Stage Specific RNA Transcription in Developing Sea Urchins and Their Chromatins.  Cold Spr. Harb. Symp. Quant. Biol. 35:629-634.
• Samis, H.V., Poccia, D.L. and Wulff, V.J. (1968).  The Effect of Salt Extraction and Heat Denaturation on the Behavior of Rat Liver Chromatin.  Biochim. Biophys. Acta 166:410-418.
  
  
  
  

	Banafshe Larijani	Teresa Barona	Scott Stephens	Erica Robbins	Christina Weidl	Gwen Wilmes	Dora Dias	Rachel Speer

Ursula Stablein	Patrick Reavey	Rui Carneiro-Martins	Michael Lukoma	Francisco Viana	Justin Herrick	Aletheia Donahue	Jake Appelbaum

Allison Schroder	Katerina Byanova	Aeri Cho	Kevin Han	Julie Fitzgerald	Michael Dowicki	Jackie Eastman	Faaiza Siddique

Ben Vincent	Sam Cutler	Darienne Myers	Peter Murphy	Burr Fong	Swapnil Mehta	Jen Kim	Crystal Edwards

Alan Wang	Michelle Huynh	Zina Chatzidimitiadou	Ashley Rivera	Emily Fan	Saumitra Thakur	Muddasir Ayaz	Anne-France Pierre

James Mooney	Karen Yeh	Stefanie Jeffrey	Aupola Kundu	Mariah Servos

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Interesting Links

• To contact the Society for Developmental Biology, click on SDB.
• To find out about the Developmental Biology of the Sea Urchin Meetings in Woods Hole,, click on Meeting.
• To contact the Sea Urchin Web Site, click on Urchins.
• To contact the Sea Urchin Harvester's Association home page click on SUHA.
• Spermatology Home Page SHP.
• The journal Human Reproduction Update HR.
• Marine Biological Lab MBL.
• Pedro's Molecular Biology Research Tools Pedro.
• Society for the Study of Fertility SSF .
• Society for the Study of Reproduction and the journal Biology of Reproduction SSR.
• Andrology Society of America Home Page ASA.
• British Andrology Society Home Page BAS.
• Nordic Society for Andrology Home Page NAS.

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