Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Winter 12-15-2018 A Mouse Model of Börjeson-Forssman-Lehmann Syndrome reveals a potential link with Autism Spectrum Disorder Cheng Cheng Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/art_sci_etds Part of the Neuroscience and Neurobiology Commons Recommended Citation Cheng, Cheng, "A Mouse Model of Börjeson-Forssman-Lehmann Syndrome reveals a potential link with Autism Spectrum Disorder" (2018). Arts & Sciences Electronic Theses and Dissertations. 1690. https://openscholarship.wustl.edu/art_sci_etds/1690 This Dissertation is brought to you for free and open access by the Arts & Sciences at Washington University Open Scholarship. It has been accepted for inclusion in Arts & Sciences Electronic Theses and Dissertations by an authorized administrator of Washington University Open Scholarship. For more information, please contact [email protected]. WASHINGTON UNIVERSITY IN ST. LOUIS Division of Biology and Biomedical Sciences Developmental, Regenerative and Stem Cell Biology Dissertation Examination Committee: Azad Bonni, Chair Joseph Dougherty Harrison Gabel Jason Yi Andrew Yoo A Mouse Model of Börjeson-Forssman-Lehmann Syndrome reveals a potential link with Autism Spectrum Disorder By Cheng Cheng A dissertation presented to The Graduate School of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Dec. 2018 St. Louis, Missouri © 2018, Cheng Cheng Table of Contents List of Figures and Tables ............................................................................................................ v Acknowledgments ...................................................................................................................... viii Abstract of the Dissertation ........................................................................................................ ix Chapter 1: Pathogenesis of Börjeson -Forssman-Lehmann syndrome: insights from PHF6 function .......................................................................................................................................... 1 Abstract ....................................................................................................................................... 2 X-linked Intellectual disability is a prevalent neurodevelopmental disorder .............................. 3 Börjeson-Forssman-Lehmann syndrome (BFLS) ....................................................................... 6 Mechanisms of PHF6 function .................................................................................................... 8 PHF6 in BFLS Pathogenesis ..................................................................................................... 12 Perspectives ............................................................................................................................... 14 Acknowledgments ..................................................................................................................... 27 Author Contributions ................................................................................................................ 27 Chapter 2: A Mouse Model of Börjeson -Forssman-Lehmann syndrome displays cognition defects and neural hyperexcitability ......................................................................................... 28 Abstract ..................................................................................................................................... 29 Introduction ............................................................................................................................... 30 Results ....................................................................................................................................... 32 Generation of patient-specific PHF6 mutant mouse model of BFLS ............................ 32 PHF6 C99F knock-in mice displays impairments in cognition, social interaction and emotionality ................................................................................................................... 33 PHF6 C99F mice exhibit incresed neuronal excitability and seizure susceptability ..... 35 Discussion ................................................................................................................................. 37 Material and Methods ................................................................................................................ 51 Plasmids ......................................................................................................................... 51 Animals .......................................................................................................................... 51 Antibodies ...................................................................................................................... 52 Immunohistochemistry ................................................................................................... 52 One-hour locomotor activity and open field behavior test ............................................. 52 ii Conditioned fear ............................................................................................................. 53 Novel object and novel location recognition .................................................................. 53 Social approach .............................................................................................................. 54 Accoustic startle reponse/prepulse inhibition ............................................................... 55 Morris water maze test ................................................................................................... 56 Seizure induction ........................................................................................................... 57 Electrophysiology ........................................................................................................... 58 Acknowledgement ..................................................................................................................... 60 Chapter 3: The Genome-Wide Functions of PHF6 in Regulating Gene Expression During Cortical Development ................................................................................................................. 61 Abstract ..................................................................................................................................... 62 Introduction ............................................................................................................................... 63 Results ....................................................................................................................................... 65 PHF6 regulated neurogenic and synaptic genes during brain development .................. 65 Transcriptomic comparisons between PHF6 knockout and C99F knock-in cortex ....... 67 PHF6-regulated genes are implicated in neurodevelopmental disorder of cognition ... 68 Mutation of PHF6 in a patient with BFLS and autism spectrum disorder ..................... 69 Discussion ................................................................................................................................. 72 Material and Methods ................................................................................................................ 82 RNA-seq ......................................................................................................................... 82 Permutation testing ......................................................................................................... 82 Gene enrichment analyses .............................................................................................. 82 Brainspan developmental analyses ................................................................................. 83 Biocytin injection and post-hoc staining ........................................................................ 84 Statistical analyses .......................................................................................................... 84 Ackowledgement ....................................................................................................................... 85 Chapter 4: Conclusion and Future Directions ......................................................................... 86 Significance ............................................................................................................................... 87 Future Directions ....................................................................................................................... 88 The biological consequence of PHF6 in regulating neural maturation ......................... 88 What is the molecular mechanisms underlying BFLS pathogenesis ............................ 89 Epigenetic regulation of transcription by PHF6 ............................................................. 90 iii Is entorhincal cortex-hippocampal pathway disrupted in mouse model of BFLS ........ 92 How do entorhinal cortex layer II stellate neurons function during contextual fear learning ..................................................................................................................................... 92 Altered neuronal and circuits excitability in BFLS ......................................................