Department of Pharmacology : Frederick E. Williams

Institutions/Degrees

Research Interests
My laboratory focuses on eliciting the molecular mechanisms of learning and memory and relating these findings to childhood learning disorders. Specifically, I have developed an animal model of spatial learning and memory using the zebrafish. My interest is in using this model to identify molecular problems that could lead to a learning disorder. In addition, mental retardation and other developmental disabilities are disorders that can be caused by pharmacological or environmental exposure to chemicals. Therefore, my interests have expanded to not only molecular mechanisms of learning, but how those mechanisms are disrupted, including by chemical means.

The zebrafish is a convenient and exciting animal being used in research today. Many laboratories use this animal to study development of organs, as do we. Our developmental research has focused on two chemicals and their related compounds. Dichloroacetic acid is a chlorination by-product in treated water. We have exposed animals to produce acute toxicities in development (J Biochem Mol Toxicol 19(1):52-58), and we will continue researching other aspects of the toxicity of compound. Further pursuit of the toxicities of this compound has lead to a second publication using ellagic acid to ameliorate the effects of DCA exposure (J Biochem Mol Toxicol, 20(4): 183-190, 2006). I hope to continue my collaboration with Ezdihar Hassoun, Ph.D. in this regard.

Thimerosal is a mercury containing compound found in health related items. Our work in this field has identified acute toxicities at low levels of exposure in developing zebrafish. Additionally, we are currently using a transgenic zebrafish, AB-Hsp70/4-GFP, developed in John Kuwada's laboratory at the University of Michigan, to assess more accurately which organs are involved in these toxicities. Cell death in these organs is then verified through other means. We will continue our research in the field of mercury exposure as it relates to development, learning, memory, and disorders of these. This work has been presented at numerous meetings and has recently been submitted for publication.

In the learning and memory field, our focus is the use of behavioral tasks to ascertain if the zebrafish has behavioral problems due to chemical or other exposures. We developed a learning and memory test for zebrafish (Williams et al., Behavioural Processes 58: 125-132). The data from this paper provided a baseline for comparison to groups of animals that learn the task faster or slower based on experimental exposure to chemicals. This was the thrust of the behavioral studies in "Ethanol effects on the developing zebrafish: neurobehavior and skeletal morphogenesis", published with collaborator Mike Carvan from the University of Wisconsin/ Milwaukee (Neurotox. teratol. 26: 757-768). Other studies in this area involve MeHg, learning, and Selenium amelioration, with collaborator Dan Weber from the Great Lakes WATER Institute in Milwaukee. This data has been recently submitted to Nanotoxicology and teratology.

One currently funded research topic involves developing a screen for skin cancer using the zebrafish and using it in high throughput to find chemicals that could treat or prevent cancer. The aim here is to develop techniques that shorten the timeframe of skin cancer development from months to weeks in an effort to look at chemicals that have an effect on cancer development. Another funded project currently going on in the laboratory together with the Liu laboratory is examining sulfation of proteins and xenobiotics in zebrafish. The Liu laboratory has several sulfating enzymes cloned from the fish, and the aim of the project is to find out what they do and more importantly, what the proteins they sulfate do. This will be accomplished partially through knockdown experiments in developing zebrafish.

Other current projects are: the measurement of mRNA levels to identify messages in the brain that are induced or reduced in expression due to learning the spatial alternation task above or specific toxins, creating a transgenic zebrafish model expressing mutant forms of the muscarinic acetylcholine M1 receptor and analyzing animals that express these mutant forms in the developing nervous system, developing a small mutagenesis screen that can be used to identify mutations in animals that fail to learn the task above chance levels, nanotoxicology of metal based materials, and finally examining children that have learning disabilities (specifically autism) with regard to their exposure to environmental pollutants.

Current Memberships in Professional Societies:
Sigma Xi - member 2001 — present
American Association of Colleges of Pharmacy — member 2002 - present

Peer-Reviewed Papers

1. Saffran, M., G. S. Kumar, C. Savariar, J.C. Burnham, F. Williams and D. C. Neckers. A new approach to the oral administration of Insulin and other peptide drugs, Science, 233, pp. 1081- 84, 1986.
2. Saffran, M., G. S. Kumar, D. G. Neckers, F. Williams, J. B. Field, R. H. Jones and L. Panepinto. New approaches to oral administration of peptide drugs, Pharmaceutisch Weekblad Scientific Edition, 10, pp. 43-44, 1988.
3. Williams, F. E. and R. J. Trumbly. Characterization of TUP1, a Mediator of glucose repression in Saccharomyces cerevisiae, Molecular and Cellular Biology, 10, pp. 6500-6511, 1990.
4. Williams, F. E., U. Varanasi, and R. J. Trumbly. The Cyc8 and TUP1 proteins involved in glucose repression in yeast are associated in a protein complex. Molecular and Cellular Biology, 11, pp. 3307-3316, 1991.
5. Freisheim, J. H., M. Ratnam, T. P. McAlinden, K. M. R. Prasad, F. E. Williams, G. R. Westerhof, J. H. Schornagel and G. Jansen. Molecular events in the membrane transport of methotrexate in human CCRF-CEM leukemia cell lines, Advances in Enzyme Regulation, 32, pp. 17-31, 1992.
6. Williams, F. E., M. Ratnam, T. P. McAlinden, G. Jansen, J. H. Schornagel, and J. H. Freisheim. Transformation of an L-cell line with the DNA coding for the reduced-folate / methotrexate transporter protein from a CCRF-CEM human leukemia cell line, Advances in Experimental Medicine and Biology: Chemistry and Biology of Pteridines and Folates (J. E. Ayling, M. G. Nair and C. M. Baugh, eds.) Plenum Press, New York, pp. 779-782, 1993.
7. Williams, F. E. and M. Saffran. Insulin in the drinking water of rats, Drug Delivery, 3, pp. 81-85, 1996.
8. Saffran, M., B. Pansky, G. C. Budd and F. E. Williams. Insulin and the gastrointestinal tract, Journal of Controlled Release 4, pp. 89-98, 1997.
9. Huang, X. P., S. Peseckis, F. Williams, and W. S. Messer, Jr. Pharmacological characterization ion of human M1 muscarinic acetylcholine receptors with double mutations at the junction of TM VI and the third extracellular domain. J. Pharmacol. Exp. Therap. 286: 1129-1139, 1998.
10. Huang, X. P., P. I. Nagy, F. E. Williams, S. M. Peseckis, and W. S. Messer, Jr. Role of threonine 192 and asparagine 382 in agonist and antagonist interactions with M₁ muscarinic receptors. Br. J. Pharmacol. 126(3): 735-745, 1999.
11. Huang, X. P., F. E. Williams, S. M. Peseckis, and W. S. Messer, Jr. Differential modulation of agonist potency and receptor coupling by mutations of Ser388Tyr and Thr389Pro at the junction of transmembrane domain VI and the third extracellular loop of human M1 muscarinic acetylcholine receptors. Mol. Pharmacol. 56: 775-783, 1999.
12. Messer, W. S., Jr., W. G. Rajeswaran, Y. Cao, H.J. Zhang, A. A. El-Assadi, C. Dockery, J. Liske, J. O'Brien, F. E. Williams, X. P. Huang, M. E. Wroblewski, P. I. Nagy, and S. M. Peseckis, Design and development of selective muscarinic agonists for the treatment of Alzheimer's disease: characterization of tetrahydropyrimidine derivatives and development of new approaches for improved affinity and selectivity for M1 receptors. Pharma. Acta Helvet. 74: 135-140, 2000.
13. Williams, F.E., White, D., and Messer, W.S. Jr., A simple spatial alternation task for assessing memory function in zebrafish. Behavioural Processes (58): 125-132, 2002.
14. Williams, F.E., Weber, D., and Carvan, M.J. III, Ethanol effects on the developing zebrafish: neurobehavior and skeletal morphogenesis. Neurotoxicology and Teratology, 26(6): 757-768, November/December 2004.
15. Williams, F.E., and Messer, W.S. Jr., Muscarinic acetylcholine receptors in the brain of the zebrafish (Danio rerio) measured by radioligand binding techniques. Comparative Biochemistry and Physiology Part C, 137: 349-353, 2004.
16. Hassoun, E., Kariya, C., and Williams, F.E., Dichloroacetate-induced developmental toxicity and production of reactive oxygen species in Zebrafish embryos. Journal of Biochemical and Molecular Toxicology, 19(1): 52-58, 2005.
17. Williams, F.E., Sickelbaugh, T., and Hassoun, E., Modulation by ellagic acid of DCA-induced developmental toxicity in the Zebrafish, (Danio rerio). Journal of Biochemical and Molecular Toxicology, 20(4): 183-190, 2006.
18. Smith, L.E., Carvan, M.J., White, D.C., Williams, F.E., and Weber, D.N., Selenomethionine attenuates early life stage abnormalities but not adult learning deficits in zebrafish developmentally exposed to methylmercury, submitted to Nanotoxicology and teratology.
19. Tomoko Yasuda, Shin Yasuda, F. E. Williams, Ming-Yih Liu, Shakhawat Bhuiyan, Rhodora Snow, Glendora Carter, and Ming-Cheh Liu. Characterization and Ontogenic Study of Novel Steroid-sulfating SULT3 Sulfotransferases from Zebrafish, Molecular and Cellular Endocrinology, in press.
20.  Liu, T.A., Bhuiyan, S., Snow, R., Yasuda, S., Yasuda, Y., Yang, Y.S., Williams, F.E., Liu, M.Y., Suiko, M., Carter, G., Liu, M.C., Identification and Characterization of Two Novel Cytosolic Sulfotransferases, SULT1-ST7 and SULT1-ST8, from Zebrafish, Aquatic Toxicology, 89: 94-102, 2008.

Selected Posters/Platform Papers Presented

1. Frederick E. Williams. Molecular approaches to learning and memory in the zebrafish, Danio rerio. Department of Medicinal and Biological Chemistry, University of Toledo, February 4, 1998. Seminar presentation.
2. W. S. Messer, Jr., F. E. Williams, P. I. Nagy, S. M. Peseckis, and X. P. Huang. Threonine and asparagine residues participate in agonist interactions with m1 muscarinic receptors. Society of Neuroscience, National meeting. New Orleans, La., November, 1998. Poster presentation.
3. F. E. Williams, and W. S. Messer, Jr., Molecular studies of learning and memory in the zebrafish, Danio rerio. Society of Neuroscience, National meeting. New Orleans, La., November, 1998. Poster presentation.
4. F. E. Williams, and W. S. Messer, Jr. Development of an alternation task as a screen for learning and memory mutants in zebrafish. Midwest Regional Zebrafish Meeting, University of Chicago, June, 1999. Poster presentation.
5. Frederick E. Williams. Spatial learning in zebrafish. Neural development and genetics of zebrafish course, Marine Biological Laboratories, Woods Hole, MA. August 1999. Seminar presentation.
6. Messer, W. S., Jr., X. P. Huang, P. I. Nagy, F. E. Williams, and S. M. Peseckis. Mutant m1 muscarinic receptors with enhanced agonist affinity and G-protein coupling. Life Sci. 64: 563-563, 1999. Poster presentation.
7. F. E. Williams. Do zebrafish think? An analysis of a spatial alternation task. Midwest Regional Zebrafish Meeting, University of Minnesota, July, 2001. Seminar presentation.
8. Williams, F.E., Zebrafish Think, What's next? Development of a spatial learning task. Invited seminar, University of Wisconsin/Milwaukee- Great Lakes WATER Institute, Milwaukee, Wisconsin, May, 2002.
9. Williams, F.E., Assessing chemically or environmentally induced developmental and cognitive changes using the zebrafish, Danio rerio. Invited seminar for faculty position, University of Toledo- College of Pharmacy, Toledo, Ohio, June, 2002.
10. Williams, F.E., and Carvan, M. J. III, Cognitive compromises: Effects of low-level lead or ethanol exposure on cognition in the zebrafish, poster presentation, The 5th International Symposium on Zebrafish genetics and development, University of Wisconsin/ Madison, June 2002.
11. Gilame,T., Williams, F.E., White, D., and Weber, D., Inorganic mercury (HgCl2) exposure to zebrafish embryos induces deficits in larval reflex response and learning, Indian Summer Festival, Milwaukee, WI, Sept. 5-7, 2003.
12. Sickelbaugh, T., Hassoun, E., and Williams, F.E., Developmental toxicities of dichloroacetic acid exposure in zebrafish embryos, Midwest Zebrafish Meeting, University of Chicago, September 12-13, 2003.
13. Williams, F.E., Preliminary developmental toxicity experiments in Danio rerio using thimerosol preservative, Midwest Zebrafish Meeting, University of Chicago, September12-13, 2003.
14. Aloia, T., and Williams, F.E., Zebrafish embryos as a model to study developmental toxicity related to thimerosol exposure, poster presentation, 6^th International Zebrafish Conference on genetics and development, University of Wisconsin, Madison, Wisconsin, 2004.
15. Renock, S., and Williams, F.E., Development of a transgenic fish line for the study of the muscarinic acetylcholine receptor hM1 in the development of Danio rerio, poster presentation, 6^th International Zebrafish Conference genetics and development, University of Wisconsin, Madison, Wisconsin, 2004.
16. Sickelbaugh, T., Hassoun, E., and Williams, F.E., Modulation by ellagic acid of DCA induced developmental defects and oxidative stress in Danio rerio, 6^th International Zebrafish Conference on genetics and development, University of Wisconsin, Madison, Wisconsin, 2004.
17. Williams, F.E., Koegle, E., Bestervelt, L, and Burmeister, W., Developmental toxicities of nanoparticles in the zebrafish, Danio rerio, 1^st International Conference on Nanotoxicology, Miami, Florida, 2006.
18. Williams, F.E., Summers, M., Vermillyea, V., Aloia, T., and Kuwada, J., Thimerosal exposure and developmental toxicities in the zebrafish, Danio rerio, 7^th International Zebrafish Conference on genetics and development, University of Wisconsin, Madison, Wisconsin, 2006.