Common use of Non Commercial Clause in Contracts

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. I hereby declare that with the exception of the RT-PCR analysis of tau exon 10 splicing in human brain in Chapter 3, all of the work presented in thesis is my own. ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want to thank my supervisor ▇▇▇▇-▇▇▇▇ ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history of the World Wars. I would also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ for encouraging me to do a PhD. Most importantly I would like to thank the NIHR BRC for funding the work in my thesis. Special thanks to ▇▇▇▇, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇▇ for helping me find my way around the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇▇▇▇▇▇ for all their help in the Lab and for helpful tips and advice. Thanks to the ▇▇▇▇▇, ▇▇▇▇▇, Petra and ▇▇▇▇▇▇▇ for technical help! I also want to thank my good friends who have been very encouraging throughout my PhD; ▇▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for their unconditional love and for always believing in me. I really appreciate you! I would like to thank my ▇▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for all their support and encouragement throughout my PhD. A special thank you goes to my boyfriend, ▇▇▇▇▇▇ ▇▇▇▇▇ for his constant love and support. I dedicate this thesis to my brother ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; thank you for everything you have done for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one of the pathological hallmarks of Alzheimer’s disease (AD). Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding protein, TDP-43 which may contribute to the clinical phenotypes observed in these cases. However TDP-43 is an RNA binding protein known to regulate alternative splicing. In addition, TDP-43 pathology has now been shown to co-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine and arginine (SR) rich proteins that either repress or promote E10 inclusion and CELF proteins are of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. I hereby declare that with the exception Download date: 04. Jun. 2023 Thesis submitted in partial fulfilment of the RT-PCR analysis degree of tau exon 10 splicing Doctorate in human brain in Chapter 3Clinical Psychology This thesis would not have been possible without an enormous amount of love, help and support from many people, so it is fitting that I begin by thanking them. First of all of the work presented in thesis is I would like to express my own. gratitude to my research supervisors: Professor ▇▇▇▇▇ ▇▇▇▇▇▇ and ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇. Both have offered unwavering support and encouragement from induction through to write up of my clinical thesis. The encouragement to pursue my own ideas combined with a collaborative supervision style has helped shaped me both as a researcher and a person. I would also like to thank ▇▇ ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want , who advised on my project, for his valuable contributions to thank my supervisor ▇▇▇▇-▇▇▇▇ ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history theoretical understanding of the World Warsarea. I would also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ for encouraging me to do a PhD. Most importantly I would like to thank the NIHR BRC for funding the work in my thesis. Special extend particular thanks to ▇▇▇▇, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇▇ for helping me find my way around the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇▇▇▇▇▇ for all their help in the Lab and for helpful tips and advice. Thanks to the ▇▇▇▇▇, ▇▇▇▇▇, Petra and ▇▇▇▇▇▇▇ for technical help! I also want to thank my good friends who have been very encouraging throughout my PhD; ▇▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇Professor ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for their unconditional love and for always believing in me. I really appreciate you! I would like to thank my ▇▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇the neuro-oncology team at King’s College Hospital for their support in identifying patients and in allowing me to attend their team meetings. I am sincerely grateful for the support from Dr ▇▇▇▇▇▇ ▇▇▇▇▇▇▇ and ▇▇ for all their support and encouragement throughout my PhD. A special thank you goes to my boyfriend, ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for his constant love and supportwith using the DEX-R questionnaire. I dedicate this thesis would like to say a very special thank you to my brother supervisors on all my clinical placements: ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; thank ▇▇, Dr ▇▇▇▇▇ ▇▇▇▇▇, Dr ▇▇▇▇▇ ▇▇▇▇, ▇▇▇▇▇ ▇▇▇▇▇▇- ▇▇▇▇▇▇▇▇, ▇▇ ▇▇▇▇▇▇ ▇▇▇▇▇, ▇▇ ▇▇▇▇▇ ▇▇▇▇▇ and ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇. Extended thanks go to those who took time to supervise and provide feedback on my case studies (▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇▇▇ and Dr ▇▇▇▇▇ ▇▇▇▇) and service evaluation project (Dr ▇▇▇▇▇ ▇▇▇▇▇). I am deeply grateful to all of them for their invaluable guidance and encouragement on placement. Thank you also to all clinicians I met in each team, who made me feel valued member of each team, rather than a trainee soon to leave. The research presented in this thesis depended on the kindness and enthusiasm of numerous participants who generously gave their time to take part in this project, and to them I am greatly indebted. I am also thankful to all the clients and their families I have worked with over the past 3 years. Each and every one of them has contributed to my learning and I feel honoured to have shared in their journeys. I would like to offer a special thanks to ▇▇ ▇▇▇ ▇▇▇▇▇▇ who was my Clinical tutor for providing me with support and encouragement throughout my time as a clinical trainee. Thank you for everything you have done your continued advice and enthusiasm over the past three years. I would also like to extend my thanks to the core course team. A cohort is only as good as its members. Here special thanks are due to my fellow trainees, for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one happy memories of the pathological hallmarks of Alzheimer’s disease (AD)many good times and their support during the more stressful. Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as As well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent the wonderful people I had around me on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding proteincourse, TDP-43 which may contribute I owe a lot to the clinical phenotypes observed wonderful people I had outside it. I am grateful to ▇▇▇▇, my Mum and brother and family and friends for their unwavering support and encouragement with balancing my continued studying and work commitments. Extended thanks go to ▇▇▇▇ for his endless love and constant reminders of a life outside work. I’d like to add a special thanks to my Dad, for his unfailing belief in these casesme. However TDP-43 is an RNA binding protein known to regulate alternative splicingHe provided unconditional love and support throughout my life and until the end of his. In additionDespite Dad not the end results, TDP-43 pathology has now been shown to comy achievements are his and we have accomplished this one together. Self-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine awareness following Traumatic Brain Injury: a systematic review of current methods of assessment, their properties and arginine (SR) rich proteins that either repress or promote E10 inclusion their correlates The Self and CELF proteins are Self-Knowledge after Frontal Lobe Neurosurgical lesions List of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20abbreviations

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. I hereby declare that with the exception Download date: 18. Jan. 2024 Thesis submitted in partial fulfilment of the RT-PCR analysis degree of tau exon 10 splicing Doctorate in human brain in Chapter 3Clinical Psychology This thesis would not have been possible without an enormous amount of love, help and support from many people, so it is fitting that I begin by thanking them. First of all of the work presented in thesis is I would like to express my own. gratitude to my research supervisors: Professor ▇▇▇▇▇ ▇▇▇▇▇▇ and ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇. Both have offered unwavering support and encouragement from induction through to write up of my clinical thesis. The encouragement to pursue my own ideas combined with a collaborative supervision style has helped shaped me both as a researcher and a person. I would also like to thank ▇▇ ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want , who advised on my project, for his valuable contributions to thank my supervisor ▇▇▇▇-▇▇▇▇ ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history theoretical understanding of the World Warsarea. I would also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ for encouraging me to do a PhD. Most importantly I would like to thank the NIHR BRC for funding the work in my thesis. Special extend particular thanks to ▇▇▇▇, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ Professor Ashkan Keyoumars and ▇▇▇▇▇ the neuro-oncology team at King’s College Hospital for helping their support in identifying patients and in allowing me find my way around to attend their team meetings. I am sincerely grateful for the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇▇▇▇▇▇ for all their help in the Lab and for helpful tips and advice. Thanks to the ▇▇▇▇▇, ▇▇▇▇▇, Petra and ▇▇▇▇▇▇▇ for technical help! I also want to thank my good friends who have been very encouraging throughout my PhD; ▇▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇from Dr ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for their unconditional love and for always believing in me. I really appreciate you! I would like to thank my ▇▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇ ▇▇▇▇ ▇▇▇▇▇▇▇▇ for all their support and encouragement throughout my PhD. A with using the DEX-R questionnaire. I would like to say a very special thank you goes to my boyfriend, ▇▇▇▇supervisors on all my clinical placements: ▇▇ ▇▇▇▇▇ for his constant love and support. I dedicate this thesis to my brother ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; thank ▇▇, Dr ▇▇▇▇▇ ▇▇▇▇▇, Dr ▇▇▇▇▇ ▇▇▇▇, ▇▇▇▇▇ ▇▇▇▇▇▇- ▇▇▇▇▇▇▇▇, ▇▇ ▇▇▇▇▇▇ ▇▇▇▇▇, ▇▇ ▇▇▇▇▇ ▇▇▇▇▇ and ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇. Extended thanks go to those who took time to supervise and provide feedback on my case studies (▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇▇▇ and Dr ▇▇▇▇▇ ▇▇▇▇) and service evaluation project (Dr ▇▇▇▇▇ ▇▇▇▇▇). I am deeply grateful to all of them for their invaluable guidance and encouragement on placement. Thank you also to all clinicians I met in each team, who made me feel valued member of each team, rather than a trainee soon to leave. The research presented in this thesis depended on the kindness and enthusiasm of numerous participants who generously gave their time to take part in this project, and to them I am greatly indebted. I am also thankful to all the clients and their families I have worked with over the past 3 years. Each and every one of them has contributed to my learning and I feel honoured to have shared in their journeys. I would like to offer a special thanks to ▇▇ ▇▇▇ ▇▇▇▇▇▇ who was my Clinical tutor for providing me with support and encouragement throughout my time as a clinical trainee. Thank you for everything you have done your continued advice and enthusiasm over the past three years. I would also like to extend my thanks to the core course team. A cohort is only as good as its members. Here special thanks are due to my fellow trainees, for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one happy memories of the pathological hallmarks of Alzheimer’s disease (AD)many good times and their support during the more stressful. Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as As well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent the wonderful people I had around me on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding proteincourse, TDP-43 which may contribute I owe a lot to the clinical phenotypes observed wonderful people I had outside it. I am grateful to ▇▇▇▇, my Mum and brother and family and friends for their unwavering support and encouragement with balancing my continued studying and work commitments. Extended thanks go to ▇▇▇▇ for his endless love and constant reminders of a life outside work. I’d like to add a special thanks to my Dad, for his unfailing belief in these casesme. However TDP-43 is an RNA binding protein known to regulate alternative splicingHe provided unconditional love and support throughout my life and until the end of his. In additionDespite Dad not the end results, TDP-43 pathology has now been shown to comy achievements are his and we have accomplished this one together. Self-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine awareness following Traumatic Brain Injury: a systematic review of current methods of assessment, their properties and arginine (SR) rich proteins that either repress or promote E10 inclusion their correlates The Self and CELF proteins are Self-Knowledge after Frontal Lobe Neurosurgical lesions List of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20abbreviations

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. Download date: 20. Jan. 2023 Centre for Robotics (CTRO) October 2022 I hereby declare that with the exception of the RT-PCR analysis of tau exon 10 splicing in human brain in Chapter 3, all of except where specific reference is made to the work presented of others, the contents of this dissertation are original and have not been submitted in whole or in part for consideration for any other degree or qualification in this, or any other University. This dissertation is the result of my own work and includes nothing which is the outcome of work done in collaboration, except where specifically indicated in the text. This dissertation contains less than 100,000 words including appendices, bibliography, footnotes, tables and equations and has less than 150 figures. This thesis is would not have been possible without the guidance and the support of several individuals. First of all, I owe my owndeepest gratitude to ▇▇. ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want to thank my supervisor ▇▇▇▇-▇▇▇▇ ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history of the World Wars. I would also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ for encouraging me to do a PhD. Most importantly I would like to thank the NIHR BRC for funding the work in my thesis. Special thanks to and ▇▇▇▇, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇. ▇▇▇▇▇ and ▇▇▇▇▇ for helping me find my way around the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇▇▇▇▇▇ for all their help in continuous support, relentless and insightful guidance. Without their invaluable contributions, patience and assistance I would not have been able to complete this study. I really thank them not only for sharing their expertise, inspiring ideas and feedbacks but also for directing and facilitating me throughout the Lab and for helpful tips and advice. Thanks course of my PhD. I would like to the ▇▇▇▇▇, ▇▇▇▇▇, Petra and ▇▇▇▇thank Professor ▇▇▇ for technical help! I also want to thank my good friends who have been very encouraging throughout my PhD; ▇▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support Scholarship and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ Memorial Foundation for their unconditional love and funding me for always believing in memy PhD. I am particularly grateful to ▇▇▇▇. I really appreciate you! I would like to thank my ▇▇▇▇▇ ▇▇▇▇▇▇▇, ▇▇▇▇▇ ▇▇▇▇-▇▇▇▇▇, and ▇▇▇▇ ▇▇▇▇▇▇ for allowing me to study with them at Osaka University. The great experience added an extra dimension to my research and helped me obtain the initial experience of the NQR system. I would like to thank Prof. ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for all their support and encouragement throughout my PhD. A special thank you goes to my boyfriend, ▇▇▇▇▇▇ ▇▇▇▇▇ for his constant love invaluable support and support. I dedicate advice in writing this thesis to my brother ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; thank you for everything you have done for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one of the pathological hallmarks of Alzheimer’s disease (AD). Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding protein, TDP-43 which may contribute to the clinical phenotypes observed in these cases. However TDP-43 is an RNA binding protein known to regulate alternative splicing. In addition, TDP-43 pathology has now been shown to co-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine and arginine (SR) rich proteins that either repress or promote E10 inclusion and CELF proteins are of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20thesis.

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. I hereby declare that with the exception Download date: 06. Oct. 2023 Thesis submitted in partial fulfilment of the RT-PCR analysis degree of tau exon 10 splicing Doctorate in human brain in Chapter 3Clinical Psychology This thesis would not have been possible without an enormous amount of love, help and support from many people, so it is fitting that I begin by thanking them. First of all of the work presented in thesis is I would like to express my own. gratitude to my research supervisors: Professor ▇▇▇▇▇ ▇▇▇▇▇▇ and ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇. Both have offered unwavering support and encouragement from induction through to write up of my clinical thesis. The encouragement to pursue my own ideas combined with a collaborative supervision style has helped shaped me both as a researcher and a person. I would also like to thank ▇▇ ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want , who advised on my project, for his valuable contributions to thank my supervisor ▇▇▇▇-▇▇▇▇ ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history theoretical understanding of the World Warsarea. I would also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ for encouraging me to do a PhD. Most importantly I would like to thank the NIHR BRC for funding the work in my thesis. Special extend particular thanks to ▇▇▇▇, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ Professor Ashkan Keyoumars and ▇▇▇▇▇ the neuro-oncology team at King’s College Hospital for helping their support in identifying patients and in allowing me find my way around to attend their team meetings. I am sincerely grateful for the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇▇▇▇▇▇ for all their help in the Lab and for helpful tips and advice. Thanks to the ▇▇▇▇▇, ▇▇▇▇▇, Petra and ▇▇▇▇▇▇▇ for technical help! I also want to thank my good friends who have been very encouraging throughout my PhD; ▇▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇from Dr ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for their unconditional love and for always believing in me. I really appreciate you! I would like to thank my ▇▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇ ▇▇▇▇ ▇▇▇▇▇▇▇▇ for all their support and encouragement throughout my PhD. A with using the DEX-R questionnaire. I would like to say a very special thank you goes to my boyfriend, ▇▇▇▇supervisors on all my clinical placements: ▇▇ ▇▇▇▇▇ for his constant love and support. I dedicate this thesis to my brother ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; thank ▇▇, Dr ▇▇▇▇▇ ▇▇▇▇▇, Dr ▇▇▇▇▇ ▇▇▇▇, ▇▇▇▇▇ ▇▇▇▇▇▇- ▇▇▇▇▇▇▇▇, ▇▇ ▇▇▇▇▇▇ ▇▇▇▇▇, ▇▇ ▇▇▇▇▇ ▇▇▇▇▇ and ▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇. Extended thanks go to those who took time to supervise and provide feedback on my case studies (▇▇ ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇▇▇ and Dr ▇▇▇▇▇ ▇▇▇▇) and service evaluation project (Dr ▇▇▇▇▇ ▇▇▇▇▇). I am deeply grateful to all of them for their invaluable guidance and encouragement on placement. Thank you also to all clinicians I met in each team, who made me feel valued member of each team, rather than a trainee soon to leave. The research presented in this thesis depended on the kindness and enthusiasm of numerous participants who generously gave their time to take part in this project, and to them I am greatly indebted. I am also thankful to all the clients and their families I have worked with over the past 3 years. Each and every one of them has contributed to my learning and I feel honoured to have shared in their journeys. I would like to offer a special thanks to ▇▇ ▇▇▇ ▇▇▇▇▇▇ who was my Clinical tutor for providing me with support and encouragement throughout my time as a clinical trainee. Thank you for everything you have done your continued advice and enthusiasm over the past three years. I would also like to extend my thanks to the core course team. A cohort is only as good as its members. Here special thanks are due to my fellow trainees, for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one happy memories of the pathological hallmarks of Alzheimer’s disease (AD)many good times and their support during the more stressful. Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as As well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent the wonderful people I had around me on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding proteincourse, TDP-43 which may contribute I owe a lot to the clinical phenotypes observed wonderful people I had outside it. I am grateful to ▇▇▇▇, my Mum and brother and family and friends for their unwavering support and encouragement with balancing my continued studying and work commitments. Extended thanks go to ▇▇▇▇ for his endless love and constant reminders of a life outside work. I’d like to add a special thanks to my Dad, for his unfailing belief in these casesme. However TDP-43 is an RNA binding protein known to regulate alternative splicingHe provided unconditional love and support throughout my life and until the end of his. In additionDespite Dad not the end results, TDP-43 pathology has now been shown to comy achievements are his and we have accomplished this one together. Self-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine awareness following Traumatic Brain Injury: a systematic review of current methods of assessment, their properties and arginine (SR) rich proteins that either repress or promote E10 inclusion their correlates The Self and CELF proteins are Self-Knowledge after Frontal Lobe Neurosurgical lesions List of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20abbreviations

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. Download date: 28. Jan. 2021 I hereby declare that with this thesis has been composed solely by myself and that it has not been submitted, in whole or in part, in any previous application for a degree. Ex- cept where stated otherwise by reference or acknowledgement, the exception work presented is entirely my own. This thesis contains fewer than 100,000 words excluding the appendices, bibliography, tables and equations. I would like to thank my wife, family and friends for all of the RT-PCR analysis of tau exon 10 splicing in human brain in Chapter 3support given to me throughout my PhD. Their support has been invaluable, all of and without them, I would not have been able to get to where I am today. I am sincerely grateful to my supervisor, ▇▇▇▇▇▇ ▇▇▇, for the guidance he has given me throughout my PhD, and for giving me the opportunity to undertake it. I would also like to thank and acknowledge his contributions to the work presented in thesis is this thesis. I would like to thank the GRChombo team [1], the COSMOS team at DAMTP, Cambridge University, and Intel for their technical support throughout my ownPhD, as well as allowing me to contribute towards their collaboration. Without them, the research carried out in this PhD would not have been possible. I would especially like to thank ▇▇▇▇ ▇▇▇▇▇▇ for guiding me when I first started my PhD, and for proofreading this work. In addition, I am grateful to ▇▇▇▇▇▇ ▇▇▇▇▇▇, ▇▇▇▇ ▇▇▇- ▇▇▇▇▇▇▇▇▇ and ▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want to thank my supervisor ▇▇▇▇-▇▇▇▇ for proofreading. In the research presented in this thesis, simulations were performed on several supercomputers: I would like to thank each centre for their technical support, and for enabling me to undertake my research. Chap. 2 is based on published research presented in [2]. I would like to thank and acknowledge the contributions of ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history of the World Wars. I would also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ , and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ to the research. I would also like to thank the University of G¨ottingen for encouraging funding my visit to see ▇▇▇▇▇ ▇ ▇ ▇▇▇▇▇ during the research process, as well as giving me the opportunity to do present my research whilst I was there. Chap. 3 and Chap. 4 are based on published research presented in [3], and on a PhD. Most importantly second paper that is currently in preparation. I would like to thank the NIHR BRC for funding the work in my thesis. Special thanks to ▇▇▇▇, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇▇ for helping me find my way around the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇▇▇▇▇▇ for all their help in the Lab and for helpful tips and advice. Thanks to the ▇▇▇▇▇, ▇▇▇▇▇, Petra and ▇▇▇▇▇▇▇ for technical help! I also want to thank my good friends who have been very encouraging throughout my PhD; ▇▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for their unconditional love the use of his initial condition code for oscillotons that was used, as well as acknowledging and for always believing in me. I really appreciate you! I would like to thank my thanking ▇▇▇▇▇▇ ▇▇▇▇▇▇, ▇▇▇▇▇▇ ▇▇▇, ▇▇▇▇▇▇ ▇.▇. ▇▇▇▇▇▇, and ▇▇▇▇▇▇▇ ▇. ▇▇▇▇ for their contributions. Chap. 5 is based on unpublished work with ▇▇▇▇▇ for all their support and encouragement throughout my PhD. A special thank you goes to my boyfriend▇▇▇▇, ▇▇▇▇▇▇ ▇▇▇▇▇ for his constant love , and support. I dedicate this thesis to my brother ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; . I would like to thank you for everything you and acknowledge their contributions towards this work. to thank every teacher and mentor I have done for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one of the pathological hallmarks of Alzheimer’s disease (AD). Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding protein, TDP-43 which may contribute to the clinical phenotypes observed in these cases. However TDP-43 is an RNA binding protein known to regulate alternative splicing. In addition, TDP-43 pathology has now been shown to co-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine and arginine (SR) rich proteins that either repress or promote E10 inclusion and CELF proteins are of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20had growing up.

Non Commercial. You may not use this work for commercial purposes. No Derivative Works - You may not alter, transform, or build upon this work. Any of these conditions can be waived if you receive permission from the author. Your fair dealings and other rights are in no way affected by the above. If you believe that this document breaches copyright please contact ▇▇▇▇▇▇▇▇▇▇▇@▇▇▇.▇▇.▇▇ providing details, and we will remove access to the work immediately and investigate your claim. Download date: 14. Oct. 2020 Firstly, I’d like to thank ▇▇▇▇▇, ▇▇▇, Dad, ▇▇▇▇▇▇ (the miniature poodle) and Inca (the labrador). Your love has been without condition and I hereby declare that with the exception of the RT-PCR analysis of tau exon 10 splicing in human brain in Chapter 3, all of the work presented in thesis is am very proud to have such wonderful people (and dogs) close to me. I’d like to thank my ownsupervisor ▇▇▇▇. ▇▇▇▇ ▇▇▇▇▇▇ who has inspired my passion for physics and its applications. ▇▇▇▇ has made the last 3.5 years very enjoyable and has given me the confidence to succeed in the next phase of my life. I would also like to thank Dr. ▇▇▇▇ ▇▇▇▇▇▇▇▇ October 2011 Firstly I want to thank my supervisor ▇▇▇▇-▇▇▇▇ ▇▇▇▇▇ for all his guidance, support and encouragement over the last three years of my PhD. ▇▇▇▇-▇▇▇▇ has invested a lot of time in me to which I am extremely grateful. I am also thankful for all the personal advice and the amazing conversations regarding issues arising around the world, French culture and the history of the World Warsspent a memorable summer visiting him at LLNL where I learnt invaluable lessons on how to conduct research. I would I’d also like to thank my second supervisor ▇▇▇▇▇ ▇▇▇▇▇▇ for supportive discussions and advice throughout my PhD. I thank my undergraduate tutors; ▇▇▇▇▇▇▇ ▇’▇▇▇▇▇▇, ▇▇▇▇▇▇▇ ▇▇▇▇▇ and ▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇ for encouraging me to do a PhD. Most importantly I would like to thank the NIHR BRC for funding the work in my thesis. Special thanks to ▇▇▇▇, Tigany, ▇▇▇▇▇▇, ▇▇▇▇▇, ▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇▇ for helping me find my way around the laboratory in the first few months and for teaching me some of the very useful molecular biology techniques I have picked up. Thank you to Amr for his help with analysis of my results and ▇▇▇▇▇ for all the helpful advice in the Lab. Also, I would like to thank ▇▇▇ and ▇▇- ▇▇▇▇▇▇ for else who has been a part of ▇▇▇▇’s research group over the last few years. It has been a pleasure to work with all their help in the Lab and for helpful tips and adviceof you. Thanks I would also like to the ▇thank ▇▇▇▇, ▇▇▇ and ▇▇▇, Petra and ▇▇ for always being willing to discuss science even with a pint in hand. And of course ▇▇▇▇▇▇ for technical help! I also want whose general discomfort whilst being in Los Angeles was a great form of relief. And to thank the rest of my good friends friends, notably ▇▇▇▇▇ and ▇▇▇ who have been very encouraging throughout my PhD; lived with me during this PhD. Although ▇▇▇▇▇ should be thanking me. A special mention goes to ▇▇▇▇▇, whose own career in physics was inspirational. To everyone at Dulwich, particularly ▇▇▇▇▇▇▇, ▇▇▇▇▇Q, ▇▇▇, ▇▇▇▇▇ and especially ▇▇▇▇▇ ▇▇▇▇▇▇. Thanks to my Aunty ▇▇▇▇▇▇ for all her support and prayers as well as the lovely Sunday lunches. Special thanks go to my Mother, ▇▇▇ ▇.▇ ▇▇▇▇▇▇▇▇ and Father, ▇▇▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for their unconditional love and for always believing in me. I really appreciate you! I would like to thank my ▇▇▇▇▇▇▇▇, ▇▇▇▇▇▇▇ and ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇ for all their support , who prove that cricket is not just about a rig and encouragement throughout my PhD. A special thank you goes to my boyfriend, ▇▇▇▇▇▇ ▇▇▇▇▇ for his constant love and supportpipes. I dedicate Publications‌ The following publications are mentioned in this thesis to my brother ▇▇▇▇▇▇▇ ▇▇▇▇▇▇▇▇; thank you for everything you have done for our family. Neurofibrillary tangles consist of hyperphosphorylated tau, one of the pathological hallmarks of Alzheimer’s disease (AD). Alternative splicing of the tau pre-mRNA produces six tau isoforms with or without exon 10 (E10); tau4R or tau3R, microtubule binding repeats respectively. In normal human brains the ratio of 4R/3R tau is approximately one. Aberrant E10 splicing is also observed in some sporadic AD cases as well as Pick’s disease. The rationale for this study is that abnormal splicing may predispose to neurological diseases such as AD either through abnormal expression and/or activity of splicing factors that control tau alternative splicing. In particular, expression levels of splicing factors regulating tau pre-mRNA splicing may be altered in AD and contribute to pathogenesis. The AD model used was dependent on the presence or absence of TDP43 inclusions because approximately 30 % of AD cases have inclusions of the RNA binding protein, TDP-43 which may contribute to the clinical phenotypes observed in these cases. However TDP-43 is an RNA binding protein known to regulate alternative splicing. In addition, TDP-43 pathology has now been shown to co-occur with tau pathology in some tauopathies. Whether the TDP-43 inclusions in AD is incidental or whether it contributes to more severe clinical phenotype remains unresolved. This model was used to determine if TDP43 pathology in AD exacerbates the changes in splicing factor expression in AD cases with TDP43 pathology. Splicing factors including serine and arginine (SR) rich proteins that either repress or promote E10 inclusion and CELF proteins are of particular interest. The brain selective CELF3 promotes tau E10 inclusion in vitro by binding to an intronic element in tau pre-mRNA. The levels of expression of CELF proteins and SR proteins that modulate tau splicing were characterised in five brain regions (frontal cortex, temporal cortex, amygdala, hippocampus and cerebellum) using post-mortem brain tissue from AD patients. Tau E10 alternative splicing pattern was also analysed in these brain regions. We investigated the expression levels of CELF proteins (CELF1, CELF3 and CELF4) and SR proteins (SC35, SRp40, and SRp55) in AD cases with (+) and without (-) TDP-43 inclusions compared to aged-matched controls. Tau E10 alternative splicing pattern analysis revealed that tau4R was increased in the amygdala and hippocampus of AD brain. We found, by quantitative RT-PCR, that the expression level of CELF3 RNA is increased in the amygdala and frontal cortex of AD cases, regardless of TDP-43 inclusions. Interestingly, in the amygdala of ADTDP43- cases where an increase in tau4R is found, we also found an increased expression of CELF4, SRp40 and SRp55 RNA. Although CELF3, CELF4 and SRp40 promote tau E10 inclusion in vitro, SRp55 inhibits tau E10 inclusion in vitro; whether this occurs in vivo is unknown. The association of abnormal expression of SR proteins and CELF proteins as well as aberrant tau E10 splicing in brain regions affected during AD pathogenesis may be a contributing factor to disease. Independent of the role of SR proteins and CELF proteins in sporadic AD, these tau splicing factors may be therapeutic targets to correct aberrant tau splicing in tauopathies. Future work will determine whether these splicing factors (CELF3, CELF4, SRp40 and SRp55) show abnormal expression in other neurodegenerative diseases. DECLARATION 2 ACKNOWLEDGEMENT 3 ABSTRACT 4 TABLE OF CONTENTS 6 LIST OF TABLES 11 LIST OF FIGURES 12 ABBREVIATIONS 15 PUBLICATIONS ARISING FROM THIS THESIS 19 1 Tau splicing factors in neurodegenerative diseases 20 1.1 Introduction 20thesis