Advertisement

Achieving Robustness to Confirm Controversial Hypotheses: A Case Study in Cell Biology

  • Emiliano TrizioEmail author
Chapter
Part of the Boston Studies in the Philosophy of Science book series (BSPS, volume 292)

Abstract

Recent developments in cellular microscopy provide an interesting example of the role played by what William Wimsatt calls “robustness analysis” in the establishment of experimental results. According to a commonly accepted biochemical model, clathrin-mediated endocytosis (that is, one of the main processes by which external material is internalized by the cell via the plasma membrane) takes place only if the size of the entering object does not exceed 120–150 nm. However recent studies provide evidence that invasive bacteria whose diameter is far larger than 150 nm can enter host cells in a clathrin-dependent manner. In particular, images obtained by fluorescence microscopy indicate the presence of clathrin molecules and their active role in such processes. Yet, due to the well-known risk that artifacts introduced during the preparation of the sample may influence the results obtained with this technique, the scientific community still does not deem the currently available evidence sufficient to revise the well-established and so far unchallenged model of clathrin-mediated endocytosis. The aim of ongoing research is thus to crosscheck the results of fluorescence microscopy by means of techniques involving transmission electron microscopy. The focus of this chapter will be on the methodologies adopted in a type of “correlative microscopy” combining (cryo-) fluorescence microscopy and (cryo-) electron tomography. It will be argued that real cases of robustness analysis offer, in general, a complicated pattern in which, a multiplicity of derivations are indeed combined, but their independence comes in degree and the results they yield stand with one another in a relation of partial overlap rather than identity. It will thus appear that the situation portrayed by Wimsatt’s robustness scheme is often to be regarded as an aim to be pursued through a long and stepwise process or even as a regulative ideal directing the researchers’ efforts, rather than as a readily available option in their methodological tool-box.

Keywords

Theoretical Principle Bacterial Invasion Chemical Fixation Joint Implementation Correlative Microscopy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

Writing this chapter would have been impossible without the keen help of Dr. Anna Sartori Rupp of the Institut Pasteur (Paris), who has patiently introduced me to the current state of the research on endocytosis and whose suggestions have constantly guided my work. I wish also to thank Léna Soler and Hubertus Nederbragt for helping me to improve this article.

References

  1. Betzig, E., et al. 2006. “Imaging Intracellular Fluorescent Proteins at Nanometer Resolution.” Science 313:1642–5.CrossRefGoogle Scholar
  2. Cheng, Y., et al. 2007. “Cryo-Electron Tomography of Clathrin-Coated Vesicles: Structural Implications for Coat Assembly.” Journal of Molecular Biology 365:892–9.CrossRefGoogle Scholar
  3. Conner, S.D., and S.L. Schmid. 2003. “Regulated Portals of Entry into Cell.” Nature 422:37–44.CrossRefGoogle Scholar
  4. Ehrkich, M., et al. 2004. “Endocytosis by Random Initiation and Stabilization of Clathrin-Coated Pits.” Cell 118:591–605.CrossRefGoogle Scholar
  5. Hacking, I. 1985. “Do We See Through a Microscope?” In Images of Science, edited by P.M. Churchland and C.A. Hooker, 132–52. Chicago and London: The university of Chicago Press.Google Scholar
  6. Keen, J.H. 1987. “Clathrin Assembly Proteins: Affinity Purification and a Model for Coat Assembly.” The Journal of Cell Biology 105:1989–98.CrossRefGoogle Scholar
  7. Kelly, B. 1997. “Is Dynamin Really a ‘Pinchase’?” Trends in Cell Biology 7:257–9.CrossRefGoogle Scholar
  8. Marsh, M., and H.T. McMahon. 1999. “The Structural Era of Endocytosis.” Science 285:215–20.CrossRefGoogle Scholar
  9. McMahon, H.T. 1999. “Endocytosis: An Assembly Protein for Clathrin Cages.” Current Biology 9:R332–5.CrossRefGoogle Scholar
  10. Nederbragt, H. 2003. “Strategies to Improve the Reliability of a Theory: The Experiment of Bacterial Invasion into Cultured Epithelial Cells.” Studies in History and Philosophy of Biological and Biomedical Sciences 34:593–614.CrossRefGoogle Scholar
  11. Nickles, T. 1989. “Justification and Experiment.” In The Uses of Experiment. Studies in the Natural Sciences, edited by D. Gooding, T. Pinch, and S. Schaffer, 299–333. Cambridge: Cambridge University Press.Google Scholar
  12. Veiga, E., and P. Cossart. 2005. “Listeria Hijacks the Clathrin-Dependent Endocytic Machinery to Invade Mammalian Cells.” Nature Cell Biology 7(9):894–900.CrossRefGoogle Scholar
  13. Veiga, E., and P. Cossart. 2006. “The Role of Clathrin-Dependent Endocytosis in Bacterial Internalization.” Trends in Cell Biology 16(10):499–504.CrossRefGoogle Scholar
  14. Veiga, E., et al. 2007. “Invasive and Adherent Bacterial Pathogens Co-Opt Host Clathrin for Infection.” Cell Host & Microbe 2:1–12.CrossRefGoogle Scholar
  15. Wimsatt, W.C. 1981. “Robustness, Reliability and Overdetermination.” In Scientific Inquiries and Social Sciences, edited by M.B. Brewer and B.E. Collins 123–162. San Francisco, CA: Jossey-Bass.Google Scholar
  16. Zaremba, S., and J.H. Keen. 1983. “Assembly Polypeptides from Coated Vesicles Mediate Reassembly of Unique Clathrin Coats.” The Journal of Cell Biology 97:1339–47.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  1. 1.Archives H. PoincaréLaboratoire d’Histoire des Sciences et de Philosophie, UMR 7117 CNRSNancyFrance
  2. 2.Archives HusserlParisFrance
  3. 3.Department of PhilosophySeattle UniversitySeattleUSA

Personalised recommendations