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A Multiweek Project Examining the Chemotactic Behavior of Tetrahymena in an Undergraduate Biology Laboratory

    Authors: Rachel Hongo1, Robert T. Grammer1, Christopher E. Barton1,*
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    Affiliations: 1: Department of Biology, Belmont University, Nashville, TN 37212
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Received 02 April 2019 Accepted 16 January 2020 Published 28 February 2020
    • ©2020 Author(s). Published by the American Society for Microbiology
    • [open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.

    • Supplemental materials available at http://asmscience.org/jmbe
    • *Corresponding author. Mailing address: Department of Biology, Belmont University, 1900 Belmont Boulevard, Nashville, TN 37212. Phone: 615-460-5714. E-mail: [email protected].
    Source: J. Microbiol. Biol. Educ. February 2020 vol. 21 no. 1 doi:10.1128/jmbe.v21i1.1805
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    Abstract:

    A multi-week laboratory project has been developed to incorporate elements of student investigation, chemotactic behavior of protists, and genetic effects on chemotactic activity of Tetrahymena.  The chemotaxis assay is based on spectrophotometric detection of protist-induced light scattering as cells migrate into a density gradient containing a known attractant. The three-week project consists of an introductory chemotaxis assay, investigation of dose-response effects, and culminates with the exploration of a Tetrahymena genetic mutant with known defects in motility.  Additionally, this project incorporates a microscopic investigation of cellular structure and swimming behavior of mutant and wild-type cells.  Students have responded well to the nature of the project, displaying clear understanding of the mechanism of the assay as well as the response of the protists to environmental manipulation and the molecular defects in the mutant cells.

References & Citations

1. Smith JJ, Wiley EA, Cassidy-Hanley DM 2012 Tetrahymena in the classroom Methods Cell Biol 109 411 430 10.1016/B978-0-12-385967-9.00016-5 22444155 3587665 http://dx.doi.org/10.1016/B978-0-12-385967-9.00016-5
2. Bozzone DM 2005 Using microbial eukaryotes for laboratory instruction and student inquiry 81 109 O’Donnell MA Tested studies for laboratory instruction 26 Proceedings of the 26th Workshop/Conference of the Association for Biology Laboratory Education
3. Sauvant MP, Pepin D, Piccinni E 1999 Tetrahymena pyriformis: a tool for toxicological studies. A review Chemosphere 38 7 1631 1669 10.1016/S0045-6535(98)00381-6 10070737 http://dx.doi.org/10.1016/S0045-6535(98)00381-6
4. Láng J, Kőhidai L 2012 Effects of the aquatic contaminant human pharmaceuticals and their mixtures on the proliferation and migratory responses of the bioindicator freshwater ciliate Tetrahymena Chemosphere 89 5 592 601 10.1016/j.chemosphere.2012.05.058 22698373 http://dx.doi.org/10.1016/j.chemosphere.2012.05.058
5. Rissing SW, Cogan JG 2009 Can an inquiry approach improve college student learning in a teaching laboratory? CBE Life Sci Educ 8 55 61 10.1187/cbe.08-05-0023 19255136 2649651 http://dx.doi.org/10.1187/cbe.08-05-0023
6. Knutson K, Smith J, Nichols P, Wallert MA, Provost JJ 2010 Bringing the excitement and motivation of research to students. Using inquiry and research-based learning in a year-long biochemistry laboratory. Part II. Research-based laboratory—a semester-long research approach using malate dehydrogenase as a research model Biochem Mol Biol Educ 38 5 324 329 10.1002/bmb.20401 http://dx.doi.org/10.1002/bmb.20401
7. Lentz TB, Ott LE, Robertson SD, Windsor SC, Kelley JB, Wollenberg MS, Dun RR, Goller CC 2017 Unique down to our microbes—assessment of an inquiry-based metagenomics activity J Microbiol Biol Educ 18 2 doi: https://doi.org/10.1128/jmbe.v18i2.1284 10.1128/jmbe.v18i2.1284 5576765 http://dx.doi.org/10.1128/jmbe.v18i2.1284
8. Deutch C 2007 Degradative enzymes from the pharmacy or health food store: interesting examples for introductory biology laboratories Am Biol Teach 69 6 e64 e70 10.1662/0002-7685(2007)69[64:DEFTPO]2.0.CO;2 http://dx.doi.org/10.1662/0002-7685(2007)69[64:DEFTPO]2.0.CO;2
9. Grammer RT 2012 Quantitation & case-study-driven inquiry to enhance yeast fermentation studies Am Biol Teach 74 6 414 420 10.1525/abt.2012.74.6.10 http://dx.doi.org/10.1525/abt.2012.74.6.10
10. Koppelhus U, Hellung-Larsen P, Leick P 1994 An improved quantitative assay for chemokinesis in Tetrahymena Biol Bull 187 8 15 10.2307/1542160 7918798 http://dx.doi.org/10.2307/1542160
11. Attwell JG, Bricker CS, Schwandt A, Gorovsky MA, Pennock DG 1992 A temperature-sensitive mutation affecting synthesis of outer arm dyneins in Tetrahymena thermophila J Protozool 39 2 261 266 10.1111/j.1550-7408.1992.tb01312.x 1533674 http://dx.doi.org/10.1111/j.1550-7408.1992.tb01312.x

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2020-02-28
2020-07-03

Abstract:

A multi-week laboratory project has been developed to incorporate elements of student investigation, chemotactic behavior of protists, and genetic effects on chemotactic activity of Tetrahymena.  The chemotaxis assay is based on spectrophotometric detection of protist-induced light scattering as cells migrate into a density gradient containing a known attractant. The three-week project consists of an introductory chemotaxis assay, investigation of dose-response effects, and culminates with the exploration of a Tetrahymena genetic mutant with known defects in motility.  Additionally, this project incorporates a microscopic investigation of cellular structure and swimming behavior of mutant and wild-type cells.  Students have responded well to the nature of the project, displaying clear understanding of the mechanism of the assay as well as the response of the protists to environmental manipulation and the molecular defects in the mutant cells.

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Figures

Image of FIGURE 1

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FIGURE 1

chemotaxis toward various concentrations of proteose peptone, a known chemoattractant. Students performed the described experiment at room temperature in a Genesys 20 spectrophotometer according to the instructions provided in the Week One student protocol (described in Appendix 1 ).

Source: J. Microbiol. Biol. Educ. February 2020 vol. 21 no. 1 doi:10.1128/jmbe.v21i1.1805
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Image of FIGURE 2

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FIGURE 2

Chemotaxis of toward increasing concentrations of proteose peptone. Students carried out a chemotaxis assay utilizing and increased concentrations of proteose peptone (described in Appendix 2 ). Results varied as to whether 4 mg/mL or 8 mg/mL gave the larger response, but 12 mg/mL consistently gave the poorest chemotaxis. Students were encouraged to discuss possible reasons for that effect.

Source: J. Microbiol. Biol. Educ. February 2020 vol. 21 no. 1 doi:10.1128/jmbe.v21i1.1805
Download as Powerpoint
Image of FIGURE 3

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FIGURE 3

The effect of gene mutation on the chemotaxis of Student-generated data examining the chemotactic ability of mutants as compared with wild-type cells (described in Appendix 3 ). Data show that cells lacking a functional gene display a significant decrease in their ability to migrate toward a chemoattractant in this assay.

Source: J. Microbiol. Biol. Educ. February 2020 vol. 21 no. 1 doi:10.1128/jmbe.v21i1.1805
Download as Powerpoint

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