Seizure and Behavioral Phenotyping of the Scn1a Mouse Model of Ge

Georgia State University ScholarWorks @ Georgia State University Nursing Dissertations (PhD) School of Nursing 12-7-2012 Seizure and Behavioral Phenotyping of the Scn1a Mouse Model of Genetic Epilepsy with Febrile Seizures Plus Ashley W Helvig Follow this and additional works at: https://scholarworks.gsu.edu/nursing_diss Recommended Citation Helvig, Ashley W., "Seizure and Behavioral Phenotyping of the Scn1a Mouse Model of Genetic Epilepsy with Febrile Seizures Plus." Dissertation, Georgia State University, 2012 https://scholarworks.gsu.edu/nursing_diss/32 This Dissertation is brought to you for free and open access by the School of Nursing at ScholarWorks @ Georgia State University It has been accepted for inclusion in Nursing Dissertations (PhD) by an authorized administrator of ScholarWorks @ Georgia State University For more information, please contact scholarworks@gsu.edu ACCEPTANCE This dissertation, SEIZURE AND BEHAVIORAL PHENOTYPING OF THE SCN1A MOUSE MODEL OF GENETIC EPILEPSY WITH FEBRILE SEIZURES PLUS, by Ashley W Helvig was prepared under the direction of the candidate’s dissertation committee It is accepted by the committee members in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Nursing in the Byrdine F Lewis School of Nursing and Health Professions, Georgia State University Michael J Decker PhD, RN, RRT, D.ABSM Committee Chairperson Shih-Yu Lee PhD, RNC Committee Member Andrew Escayg, PhD Committee Member Toni Whistler, PhD Committee Member _ Date This dissertation meets the format and style requirements established by the Byrdine F Lewis School of Nursing and Health Professions It is acceptable for binding, for placement in the University Library and Archives, and for reproduction and distribution to the scholarly and lay community by University Microfilms International _ Joan Cranford, EdD, RN Assistant Dean for Nursing Byrdine F Lewis School of Nursing and Health Professions _ Margaret C Wilmoth, PhD, MSS, RN, FAAN Dean and Professor Byrdine F Lewis School of Nursing & Health Professions AUTHOR’S STATEMENT In presenting this dissertation as a partial fulfillment of the requirements for an advanced degree from Georgia State University, I agree that the Library of the University shall make it available for inspection and circulation in accordance with its regulations governing materials of this type I agree that permission to quote from, to copy from, or to publish this dissertation may be granted by the author or, in his/her absence, by the professor under whose direction it was written, or in his/her absence, by the Assistant Dean for Nursing, Byrdine F Lewis School of Nursing and Health Professions Such quoting, copying, or publishing must be solely for scholarly purposes and will not involve potential financial gain It is understood that any copying from or publishing of this dissertation which involves potential financial gain will not be allowed without written permission from the author Ashley W Helvig i NOTICE TO BORROWERS All dissertations deposited in the Georgia State University Library must be used in accordance with the stipulations prescribed by the author in the preceding statement The author of this dissertation is: Ashley W Helvig 225 Vinestone Ct Fayetteville, GA 30215 The director of this dissertation is: Dr Michael J Decker Associate Professor Byrdine F Lewis Chair in Nursing Associate Member, Neuroscience Institute Member, Center for Behavioral Neuroscience Byrdine F Lewis School of Nursing and Health Profession Georgia State University P.O Box 3995 Atlanta, GA 30302-4019 Users of this dissertation not regularly enrolled as students at Georgia State University are required to attest acceptance of the preceding stipulations by signing below Libraries borrowing this dissertation for the use of their patrons are required to see that each user records here the information requested NAME OF USER ADDRESS DATE ii TYPE OF USE (EXAMINATION ONLY OR COPYING) VITA Ashley W Helvig ADDRESS: 225 Vinestone Ct Fayetteville, GA 30215 EDUCATION: Ph.D 2012 M.S.N 2007 B.S.N 1992 Georgia State University Atlanta, Georgia University of West Georgia Carrollton, Georgia Medical College of Georgia Augusta, Georgia PROFESSIONAL EXPERIENCE: 2012- present Assistant Professor, University of West Georgia, Carrollton, GA 2007- 2012 Associate Professor, Gordon College Barnesville, GA 2003- 2009 Staff Nurse, Charge Nurse, Piedmont Fayette Hospital Fayetteville, GA 2003 Clinical Manager, IV Team Staff Builders Home Care, Atlanta, GA 1998-2003 IV Nurse, Visiting Nurse Health System, Atlanta, GA 1995-1998 Case Manager, Central Home Health Care, Newnan, GA 1992-1995 Staff Nurse II, Egleston Children’s Hospital, Atlanta, GA 1994-1995 Home Health Nurse, PRN, Georgia Health Resources, Marietta, GA 1994 Staff Nurse, PRN, Spalding Regional Hospital, Griffin, GA PROFESSIONAL ORGANIZATIONS AND CERTIFICATIONS: 2011-2012 2006-2012 2002-2012 2001-2012 1992-2012 Georgia Association of Nurse Educators Southern Nursing Research Society National Certification -Certified Registered Nurse Infusionist National Infusion Nurses’ Society Sigma Theta Tau Nursing Honor Society HONORS: 2010 First nursing student member of the Center for Behavioral Neuroscience, Georgia State University 2006 HEART of Fayette recipient, Piedmont Fayette Hospital iii ABSTRACT SEIZURE AND BEHAVIORAL PHENOTYPING OF THE SCN1A MOUSE MODEL OF GENETIC EPILEPSY WITH FEBRILE SEIZURES PLUS by ASHLEY W HELVIG Genetic epilepsy with febrile seizures plus (GEFS+) is associated with a wide range of neurological dysfunction caused in part by limited function in voltage-gated sodium channels (Escayg & Goldin, 2010; Gambardella & Marini, 2009; Mulley et al., 2005) The seizure and behavioral phenotypes, as well as use of non-pharmacologic agents as neuroprotectants in GEFS+, are not well-understood An experimental design used an animal model of GEFS+ to explore the effects of stress on seizure phenotype, examine behavioral phenotypes, and study the effects of an omega fatty acid on abnormal behaviors noted in the various paradigms This study used C57BL/6J mice with the R1648H missense mutation on the Scn1a gene (engineered in the Escayg lab) (Martin, M S et al., 2010) The three specific aims used separate groups of animals for experimentation, and all paradigms were performed under strict laboratory conditions Data were analyzed using either an independent t-tests, two-way ANOVA or repeated measures two-way ANOVA Results showed that stress worsens seizure iv phenotype in both the Scn1aR1648H (RH) mutants and wild-type (WT) group with the RH mutants more severely impacted In addition, there was clear and consistent evidence for hyperactive locomotor behavior Lastly, no evidence was found for use of docosahexaenoic acid (DHA, an omega fatty acid) as a neuroprotectant for hyperactivity (DHA was given subcutaneously for two weeks starting at weaning) Outcomes from this study implicate that stress worsens the seizure phenotype in animals with Scn1aR1648H This study is also the first to report hyperactive locomotor behavior in animals with Scn1aR1648H Results from this study may broaden beyond GEFS+ in that we may also be able to apply the findings to other disorders with SCN1A dysfunction In addition, it may be that genetic variants affecting SCN1A, but not necessarily in epilepsy, may contribute to hyperactivity This could mean that SCN1A is a candidate gene for hyperactivity The main goal of nursing care is to reduce and prevent disease morbidity, and knowledge gained from the current study will guide clinical nursing practice, such as targeted behavioral assessment and education, as well as nursing research focusing on children with this genetic disorder v TITLE PAGE SEIZURE AND BEHAVIORAL PHENOTYPING OF THE SCN1A MOUSE MODEL OF GENETIC EPILEPSY WITH FEBRILE SEIZURES PLUS by ASHLEY W HELVIG A DISSERTATION Presented in Partial Fulfillment of Requirements for the Degree of Doctor of Philosophy in Nursing in the Byrdine F Lewis School of Nursing and Health Professions Georgia State University Atlanta, Georgia 2012 vi COPYRIGHT Copyright by Ashley W Helvig 2012 vii ACKNOWLEDGMENTS This dissertation would not have been possible without the support, encouragement, and intelligence of a multitude of people I would like to first thank Dr Michael Decker for taking me under his wing and showing me a path that I did not think was possible You have taught me so much over the past few years and stretched me as a nurse scientist Thank you so much for your inspiration and keeping me on track Your dedication will not be forgotten I would like to thank my committee members who have helped me so much through this process Thanks to Dr Toni Whistler for being so kind and pushing me to the best job I could To Dr Sylvia Lee, I thank for you not only being a wonderful teacher and support in my committee, but for inspiring greatness in research To Dr Andrew Escayg, thank you so much for agreeing to have a nurse in your lab Your willingness to help me in this process was incredible A special thank goes to Nikki Sawyer Your kindness and encouragement will not be forgotten You are a wonderful mentor and teacher To Shelly, Susan and Joy, my Ph.D buddies who have helped me survive the past four years, you are very special to me Your words of encouragement will always be with me I am extremely appreciative of the funding support through the Byrdine F Lewis School of Nursing and Health Professions as well as through the University System of Georgia that have helped to defray costs of my education throughout the past four years viii 131 Tamminga, C., Hashimoto, T., Volk, D., & Lewis, D (2004) GABA neurons in the human prefrontal cortex The American Journal of Psychiatry, 161(10), 17641764 Tang, B., Dutt, K., Papale, L., Rusconi, R., Shankar, A., Hunter, J., et al (2009) A BAC transgenic mouse model reveals neuron subtype-specific effects of a Generalized Epilepsy with Febrile Seizures Plus (GEFS+) mutation Neurobiology of Disease, 35(1), 91-102 Terra, V C., Arida, R M., Rabello, G M., Cavalheiro, E A., & Scorza, F A (2011) The utility of omega-3 fatty acids in epilepsy: more than just a farmed tilapia! Arquivos de Neuro-Psiquiatria, 69(1), 118-121 Thomas, A., Burant, A., Bui, N., Graham, D., Yuva Paylor, L., & Paylor, R (2009) Marble burying reflects a repetitive and perseverative behavior more than novelty-induced anxiety Psychopharmacology, 204(2), 361-373 Van Baar, A., Vermaas, J., Knots, E., de Kleine, M J K., & Soons, P (2009) Functioning at school age of moderately preterm children born at 32 to 36 weeks’ gestational age Pediatrics, 124(1), 251-257 Vancassel, S., Blondeau, C., Lallemand, S., Cador, M., Linard, A., Lavialle, M., et al (2007) Hyperactivity in the rat is associated with spontaneous low level of n-3 polyunsaturated fatty acids in the frontal cortex Behavioural Brain Research, 180(2), 119-126 Varvogli, L., & Darviri, C (2011) Stress management techniques: evidence-based procedures that reduce stress and promote health Health Science Journal, 5(2), 74-89 132 Voigt, R G., Llorente, A M., Jensen, C L., Fraley, J K., Berretta, M C., & Heird, W C (2001) A randomized, double-blind, placebo-controlled trial of docosahexaenoic acid supplementation in children with attention-deficit/hyperactivity disorder The Journal of Pediatrics, 139(2), 189-196 Wehmeier, P., Dittmann, R., Schacht, A., Minarzyk, A., Lehmann, M., Sevecke, K., et al (2007) Effectiveness of atomoxetine and quality of life in children with attentiondeficit/hyperactivity disorder as perceived by patients, parents, and physicians in an open-label study Journal of Child and Adolescent Psychopharmacology, 17(6), 813-830 Willis, S., Samala, R., Rosenberger, T., & Borges, K (2009) Eicosapentaenoic and docosahexaenoic acids are not anticonvulsant or neuroprotective in acute mouse seizure models Epilepsia, 50(1), 138-142 Yavin, E (2006) Versatile roles of docosahexaenoic acid in the prenatal brain: from proand anti-oxidant features to regulation of gene expression Prostaglandins, Leukotrienes and Essential Fatty Acids, 75(3), 203-211 Yuen, A W C., Sander, J., Fluegel, D., Patsalos, P., Bell, G., Johnson, T., et al (2005) Omega-3 fatty acid supplementation in patients with chronic epilepsy: a randomized trial Epilepsy & Behavior, 7(2), 253-258 Yuen, A W C., Sander, J., Flugel, D., Patsalos, P., Browning, L., Bell, G., et al (2008) Erythrocyte and plasma fatty acid profiles in patients with epilepsy: does carbamazepine affect omega-3 fatty acid concentrations? Epilepsy & Behavior, 12(2), 317-323 133 Zhang, W., Hu, X., Yang, W., Gao, Y., & Chen, J (2010) Omega-3 polyunsaturated fatty acid supplementation confers long-term neuroprotection against neonatal hypoxic-ischemic brain injury through anti-inflammatory actions Stroke, 41(10), 2341-2347 Doi: 10.1161/strokeaha.110.586081 Zobel, A., Wellmer, J., Schulze-Rauschenbach, S., Pfeiffer, U., Schnell, S., Elger, C., et al (2004) Impairment of inhibitory control of the hypothalamic pituitary adrenocortical system in epilepsy European Archives of Psychiatry and Clinical Neuroscience, 254(5), 303-311 Zuberi, S M., Brunklaus, A., Birch, R., Reavey, E., Duncan, J., & Forbes, G H (2011) Genotype-phenotype associations in SCN1A-related epilepsies Neurology, 76(7), 594-600 APPENDIX A PICROTOXIN CODING FORM 135 Mouse # Latency to Freezing/Staring (≥ 10 sec) _ 0.5) Abnormal Behavior? _ 1) Latency to 1st Myoclonic Jerk Total Number of Jerks _ Time Coded for MJ: Total Time Coded _ Minus Time Seizing Minus Latency to MJ _ Time Coded for MJ 2) Latency to 1st Focal Seizure _ 3) Latency to fully developed bilateral forelimb clonus 3.5) Latency to forelimb clonus with tonic component and twist of the body 4) Latency to GCS with no tonic component _ 4.5) Latency to GTCS w/ FLE 5) Latency to GTCS w/ FLE & HLE 5.5) Long-lasting clonus? (yes/no) 6) Death? (yes/no) Highest Level on Scale Presence of Grooming at the end? (yes/no) 136 APPENDIX B PICROTOXIN OBSERVATION FORM 137 Date: Mouse #: Myoclonic Jerk (Starting Time) Partial (P) or Generalized (G) Tonic Seizure GTCS (with bouncing) Hindlimb Extension Status Epilepticus (>15 min) Death Other/Notes: 138 APPENDIX C BEHAVIOR GRID 139 Date: Behavior Grooming Duration Grooming Frequency Scratching Rearing StretchAttend Fecal Boli 140 APPENDIX D GENOTYPING 141 bars equals RH mutant bar equals WT 142 APPENDIX E GSU IACUC APPROVAL FORM 143 144 APPENDIX F GSU IACUC RENEWAL FORM 145 146 ... dissertation, SEIZURE AND BEHAVIORAL PHENOTYPING OF THE SCN1A MOUSE MODEL OF GENETIC EPILEPSY WITH FEBRILE SEIZURES PLUS, by Ashley W Helvig was prepared under the direction of the candidate’s dissertation... targeted behavioral assessment and education, as well as nursing research focusing on children with this genetic disorder v TITLE PAGE SEIZURE AND BEHAVIORAL PHENOTYPING OF THE SCN1A MOUSE MODEL. .. degrees of phenotypes Nevertheless, 20 researchers have been and continue to use animal models of GEFS+ and report various aspects of the genotype and phenotype The Escayg laboratory has generated mouse