Western Michigan University ScholarWorks at WMU Dissertations Graduate College 4-2007 Global and Quantitative Gene Expression Analysis of the Effects of Drinking Water Exposure to Lead Acetate in Fisher 344 Male Rats Liver Worlanyo Eric Gato Western Michigan University Follow this and additional works at: https://scholarworks.wmich.edu/dissertations Part of the Chemistry Commons Recommended Citation Gato, Worlanyo Eric, "Global and Quantitative Gene Expression Analysis of the Effects of Drinking Water Exposure to Lead Acetate in Fisher 344 Male Rats Liver" (2007). Dissertations. 863. https://scholarworks.wmich.edu/dissertations/863 This Dissertation-Open Access is brought to you for free and open access by the Graduate College at ScholarWorks at WMU. It has been accepted for inclusion in Dissertations by an authorized administrator of ScholarWorks at WMU. For more information, please contact [email protected]. GLOBAL AND QUANTITATIVE GENE EXPRESSION ANALYSIS OF THE EFFECTS OF DRINKING WATER EXPOSURE TO LEAD ACETATE IN FISHER 344 MALE RATS LIVER by Worlanyo Eric Gato A Dissertation Submitted to the Faculty of The Graduate College in partial fulfillment of the requirements for the Degree of Doctor of Philosophy Department of Chemistry Dr. Jay Means, Advisor Western Michigan University Kalamazoo, Michigan April 2007 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. GLOBAL AND QUANTITATIVE GENE EXPRESSION ANALYSIS OF THE EFFECTS OF DRINKING WATER EXPOSURE TO LEAD ACETATE IN FISHER 344 MALE RATS LIVER Worlanyo Eric Gato, Ph.D. Western Michigan University, 2007 The primary objective of this research is to analyze global gene expression patterns occuring in Fisher 344 rat livers exposed to varying levels of lead and times. The hypotheses were that: 1) effects associated with Pb exposure are both dose and time dependent and 2) several genes will be over-expressed or repressed including transcripts associated with calcium signaling. Initially, the effects of Pb exposures upon morphometric indices, liver and kidney tissue histology, Pb distribution, Pb interaction with other trace metals including Zn, Cu, Co, Fe, Ni and Ca were assessed. Results showed a significant accumulation of lead in blood, liver, kidney and bone marrow in lead exposed groups with the kidney demonstrating greater damage compared to the liver. Potential interactions of calcium, iron, cobalt, copper, zinc and nickel and lead examined showed positive and negative correlation for 30 and 90 days treatment period respectively. Differentially expressed genes included genes cited in the literature and several not previously reported to be affected by lead toxicity. Expression profiles were clustered and gene ontology (GO) revealed 15 GO categories affected by chronic (90d) exposure, while 3 GO categories were affected during (30d) exposures. Pathways emphasized the importance of Pb in modulating various cellular events in a manner similar to calcium regulation, including phosphorylation and dephosphorylation, calcium Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. signaling, histone acetylation and deacetylation. Conclusions include: 1. Pb controls mammalian protein synthesis via regulating phosphorylation or dephosphorylation events of eukaryotic elongation/initiation factors 2. Pb regulates gene expression through the regulation of histone acetylases 3. Pb regulates calcium dependent transcription factor myocyte enhance factor-2 Quantitative PCR was employed in validating the microarray result and showed that Microarrays and qRT-PCR yield comparable results. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. UMI Number: 3259739 Copyright 2007 by Gato, Worlanyo Eric All rights reserved. INFORMATION TO USERS The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleed-through, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. ® UMI UMI Microform 3259739 Copyright 2007 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, Ml 48106-1346 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Copyright by Worlanyo Eric Gato 2007 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ACKNOWLEDGEMENTS I am indebted to my late grandmother Malwine Adzoa Boateng, my wife Vivian Mawuse Gato and children Worlanyo Eric Gato, Jr. and Elikem Winifred Gato, to whom I dedicate this dissertation. I want to thank the Chemistry Department of Western Michigan University for giving me the opportunity to study for a doctoral degree in Chemistry. I also want to thank Dr. Jay C. Means for allowing me to work in his laboratory and supporting me financially. I am grateful to my committee members Drs. David Huffman and Robert Eversole for their thoughtful comments and support in various ways. Also, I want to express my deep appreciation to Dr. Michael Barcelona for the various ways in which he has helped me through this program. This Chemistry program has given me the opportunity to meet and interact with people from diverse cultural, ethnic and intellectual backgrounds. I am glad to meet colleagues from Sri Lanka - Thushara Ghunasinghe, Kumar Tilak, Nayane Prasange, Piyardasha and Arana Weerasinghe, China - Minghong Liu, Wen Guo, Luchin Lin, Wend, Kenya - Wilson Okumu, David Achila, Patrick Ochieng, Joshua Muia, Paul Oshule and Joy Mwalimu; U.S. - Jenny Powell, Elizabeth Sodt and Elizabeth Semkiw, Brian Zeider and William Lizik, India - Shankar Varangati, Swapna Katram, Pramod Thekkat, Suhitha Meka and Sandhya Nair. I appreciate the friendship of these guys and a host of others. I appreciate the friendship provided by Paul Knoll and Trisha Basford my ii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Acknowledgements - Continued laboratory colleagues. Thank you, Trisha Basford for helping me to learn the techniques required for this project. Drs. David Reinhold, Susan Stapleton and Sherine Obare have provided mentoring and references for me. Pam McCartney and Annie Dobbs have been helpful in many ways. For these, I say thank you and I appreciate your kindness very much. Finally, I want to express my deep thanks to the members of Westwood Baptist Church for being a family to us. Worlanyo Eric Gato iii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. TABLE OF CONTENTS ACKNOWLEGEMENTS............................ ............................................... ii LIST OF TABLES......................................................................................... ix LIST OF FIGURES....................................................................................... x CHAPTER I. INTRODUCTION....................................................................................... 1 II. LITERATURE REVIEW........................................................................... 9 Lead Background Information ............................................................. 9 Historical Background ............................................................. 9 What is Lead?........................................................................... 11 Sources of Lead in the Environment ....................................... 12 Lead Use History ...................................................................... 14 Production, Disposal and Regulations .................................... 16 Mode of Toxicity .................................................................................. 18 Lead in Air ................................................................................ 18 Lead in Soil ............................................................................... 20 Lead in Aquatic Systems ......................................................... 22 Lead Cycling in Soils and Surface Waters ............................ 23 Human Lead Exposure Routes ............................................... 27 Lead Remediation .................................................................... 28 iv Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table of Contents - Continued CHAPTER Lead Absorption, Distribution and Excretion ......................... 29 Lead T oxicokinetics ................................................................ 35 Health Effects Associated With Pb2+ Exposure.. ............................. 42 N eurobehavioral Effects .......................................................... 42 Developmental Effects ............................................................. 44 Lead Carcinogenicity ............................................................... 46 Immunological Effects ............................................................. 48 Reproductive Effects ...............................................................