Hostname: page-component-76fb5796d-wq484 Total loading time: 0 Render date: 2024-04-25T17:25:38.747Z Has data issue: false hasContentIssue false

Confocal laser scanning microscopy method for in vivo bryozoan larvae identification

Published online by Cambridge University Press:  17 May 2010

A. Tsyganov-Bodounov*
Affiliation:
School of the Environment & Society, Biological Sciences, University of Wales, Swansea, SA2 8PP, UK
David O.F. Skibinski
Affiliation:
Institute of Life Sciences, School of Medicine, University of Wales, Swansea, SA2 8PP, UK
*
Correspondence should be addressed to: A. Tsyganov-Bodounov, School of the Environment & Society, Biological Sciences, University of Wales, Swansea, SA2 8PP, UK emails: a.tsyganov@liverpool.ac.uk, atsyganov@gmail.com
Get access

Abstract

Knowledge of bryozoan larval types is limited to a relatively small number of species. Further information is needed to establish traits of evolutionary significance and thus advance bryozoan taxonomy. Morphological classification of bryozoan larvae has previously focused on electron microscopy, epi-fluorescence microscopy and lightfield microscopy methods. These are either relatively time consuming and require fixed material, or they fail to reveal surface features of larvae. A method is presented here, based on confocal laser scanning microscopy, which allows in vivo 3-D imaging of larvae. The method could also be used for imaging other marine invertebrate larvae of comparable size.

Type
Research Article
Copyright
Copyright © Marine Biological Association of the United Kingdom 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Amos, W.B., White, J.G. and Fordham, M. (1987) Use of confocal imaging in the study of biological structures. Applied Optics 26, 32393243.Google Scholar
Cooper, M.S., Szeto, D.P., Sommers-Herivel, G., Topczewski, J., Solnica-Krezel, L., Kang, H.C., Johnson, I. and Kimelman, D. (2005) Visualizing morphogenesis in transgenic zebrafish embryos using BODIPY TR methyl ester dye as a vital counterstain for GFP. Developmental Dynamics 232, 359368.Google Scholar
Dubertret, B., Skourides, P., Norris, D.J., Noireaux, V., Brivanlou, A.H. and Libchaber, A. (2002) In vivo imaging of quantum dots encapsulated in phospholipid micelles. Science 298, 17591762.CrossRefGoogle ScholarPubMed
Giribet, G., Distel, D.L., Polz, M., Sterrer, W. and Wheeler, W.C. (2000) Triploblastic relationships with emphasis on the acoelomates and the position of Gnathostomulida, Cycliophora, Platyhelminthes, and Chaetognatha: a combined approach of 18S rDNA sequences and morphology. Systematic Biology 49, 539562.Google Scholar
Glenner, H., Hansen, A.J., Sørensen, M.V., Ronquist, F., Huelsenbeck, J.P. and Willerslev, E. (2004) Bayesian inference of the metazoan phylogeny: a combined molecular and morphological approach. Current Biology 14, 16441649.Google Scholar
Gruhl, A. (2008) Muscular systems in gymnolaemate bryozoan larvae (Bryozoa: Gymnolaemata). Zoomorphology 127, 143159.CrossRefGoogle Scholar
Gruhl, A., Wegener, I. and Bartolomaeus, T. (2009) Ultrastructure of the body cavities in Phylactolaemata (Bryozoa). Journal of Morphology 270, 306318.CrossRefGoogle ScholarPubMed
Longin, A., Souchier, C., Ffrench, M. and Bryon, P.A. (1993) Comparison of anti-fading agents used in fluorescence microscopy: image analysis and laser confocal microscopy study. Journal of Histochemistry and Cytochemistry 41, 18331840.Google Scholar
Mackey, L.Y., Winnepenninckx, B., de Wachter, R., Backeljau, T., Emschermann, P. and Garey, J.R. (1996) 18S rRNA suggests that entoprocta are protostomes, unrelated to ectoprocta. Journal of Molecular Evolution 42, 552559.Google Scholar
Mundy, S.P., Taylor, P.D. and Thorpe, J.P. (1981) A reinterpretation of phylactolaemate phylogeny. In Larwood, G.P. and Nielsen, C. (eds) Recent and fossil Bryozoa. Fredensborg: Olsen & Olsen, pp. 185190.Google Scholar
Nielsen, C. (1971) Entoproct life-cycles and the entoproct/ectoproct relationship. Ophelia 9, 209341.Google Scholar
Reed, C.G. (1977) Larval morphology and settlement of the bryozoan, Bowerbankia gracilis (Vesicularioidea, Ctenostomata): structure and eversion of the internal sac. In Chia, F.S. and Rice, M.E. (eds) Settlement and metamorphosis of marine invertebrate larvae. New York: Elsevier/North Holland Biomedical Press, pp. 4148. [Proceedings of the Symposium on Settlement and Metamorphosis of Marine Invertebrate Larvae, Toronto, Ontario, Canada.]Google Scholar
Reed, C.G. (1988) The reproductive biology of the gymnolaemate bryozoan Bowerbankia gracilis (Ctenostomata, Vesiculariidae). Ophelia 29, 123.Google Scholar
Reed, C.G. (1991) Bryozoa. In Giese, A.C., Pearse, J.S. and Pearse, V.B. (eds) Reproduction of marine invertebrates. Pacific Grove, California: The Boxwood Press, pp. 85245.Google Scholar
Reed, C.G. and Cloney, R.A. (1982) The larval morphology of the marine bryozoan Bowerbankia gracilis (Ctenostomata, Vesicularioidea). Zoomorphology 100, 2354.CrossRefGoogle Scholar
Reed, C.G., Ninos, J.M. and Woollacott, R.M. (1988) Bryozoan larvae as mosaics of multifunctional ciliary fields—ultrastructure of the sensory organs of Bugula stolonifera (Cheilostomata: Cellularioidea). Journal of Morphology 197, 127145.Google Scholar
Reed, C.G. and Woollacott, R.M. (1982) Mechanisms of rapid morphogenetic movements in the metamorphosis of the bryozoan Bugula neritina (Cheilostomata, Cellularioidea). 1. Attachment to the substratum. Journal of Morphology 172, 335348.Google Scholar
Reed, C.G. and Woollacott, R.M. (1983) Mechanisms of rapid morphogenetic movements in the metamorphosis of the bryozoan Bugula neritina (Cheilostomata, Cellularioidea). 2. The role of dynamic assemblages of microfilaments in the pallial epithelium. Journal of Morphology 177, 127143.Google Scholar
Ross, L.G. and Ross, B. (1999) Anaesthetic and sedative techniques for aquatic animals. Oxford: Blackwell Science.Google Scholar
Ryland, J.S. (1970) Bryozoans. London: Hutchinson.Google Scholar
Santagata, S. (2008a) The morphology and evolutionary significance of the ciliary fields and musculature among marine bryozoan larvae. Journal of Morphology 269, 349364.Google Scholar
Santagata, S. (2008b) Evolutionary and structural diversification of the larval nervous system among marine bryozoans. Biological Bulletin. Marine Biological Laboratory, Woods Hole 215, 323.Google Scholar
Santagata, S. and Zimmer, R.L. (2000) Comparing cell patterns of coronate bryozoan larvae with fluorescent probes. Proceedings of the 11th International Bryozoology Association Conference, Smithsonian Tropical Research Institute, Panamá.Google Scholar
Taylor, P.D. and Larwood, G.P. (1990) Major evolutionary radiations in the Bryozoa. In Taylor, P.D. and Larwood, G.P. (eds) Major evolutionary radiations. Oxford: Clarendon Press. [Systematics Association Special Volume no. 42, 209–233.]Google Scholar
Zimmer, R.L. and Woollacott, R.M. (1977) Structure and classification of gymnolaemate larvae. In Woolacott, R. and Zimmer, R.L. (eds) Biology of bryozoans. New York: Academic Press, pp. 5789.CrossRefGoogle Scholar