Macquarie University
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Investigating pathological features of a novel amytrophic lateral schlerosis mouse model

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posted on 2022-03-28, 18:44 authored by Winonah Ruby Riddell
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterised by the loss of brain and spinal cord motor neurons. A proportion of ALS patients also develop frontotemporal dementia (FTD). A mutation in the CCNF gene (CCNFS621G) was recently reported in a family with ALS and FTD, offering a new opportunity to model ALS/FTD, including development of pre-clinical animal models. Over-expression of this mutation has been studied in zebrash and in two mouse cell lines, NSC-34 and neuro-2a. However, the effects of endogenously expressed CCNFS621G on the mammalian system remain unclear. To assess this, tissues from a previously generated CRISPR-Cas9 mouse model carrying CCNFS621G in its genome (CRISPR-CCNFS621G), were studied. This thesis aimed to characterise, for the first time, pathological features in the CRISPR- CCNFS621G mouse model, to evaluate its utility for ALS research. Specifically, this study assessed the impact of CCNFS621G on cyclin F expression, and investigated characteristic ALS/FTD histopathologies (mis-localised TDP-43, hyper-phosphorylated and aggregates of TDP-43, and ubiquitinated protein inclusions) and proteomic changes within brain and spinal cord tissues from six-month-old heterozygous and homozygous CRISPR- CCNFS621G mice. Genetics, molecular biology, histology, and proteomic techniques, were employed. It was found that CCNF gene (mRNA) and cyclin F protein levels were not altered in the CRISPR-CCNFS621G mice. No characteristic ALS/FTD histopathologies were identified within brain and spinal cord tissues from the CRISPR-CCNFS621G mice, but a significant loss of spinal cord motor neurons was observed in homozygous CRISPR- CCNFS621G mice when compared to control and heterozygous CRISPR-CCNFS621G mice. Filtering and analysis of proteomics data identified aberrantly expressed proteins in the CRISPR-CCNFS621G mice, which included TBC1D15 and GNAI2 among others, that are implicated in ALS-relevant biological processes. In summary, this project systematically characterised molecular and cellular pathologies present in the CRISPR-CCNFS621G mice, which may represent early features associated with disease pathogenesis prior to the occurrence of significant motor decficits. This was the first CRISPR-CCNF mouse model study and provides insights into cellular and biochemical changes relevant to ALS/FTD linked CCNFS621G. Future studies in an older cohort are required to examine progression of these changes and to gain a full understanding of CCNF-related disease aetiology.


Table of Contents

1 Introduction -- 2 Subjects & materials -- 3 Methods -- 4 Results -- 5 Discussion.


Bibliography pages Theoretical thesis.

Awarding Institution

Macquarie University

Degree Type

Thesis MRes


MRes, Macquarie University, Faculty of Medicine and Health Sciences, Department of Biomedical Sciences

Department, Centre or School

Department of Biomedical Sciences

Year of Award


Principal Supervisor

Shu Yang

Additional Supervisor 1

Ian Blair


Copyright Winonah Ruby Riddell 2020




1 online resource (xviii, 141 pages) : illustrations

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