Project ID NS-MH2024_25


Co Supervisor 1A Institute of Psychiatry, Psychology & Neuroscience, School of Neuroscience, Department of Basic & Clinical NeuroscienceWebsite

Co Supervisor 1B Institute of Psychiatry, Psychology & Neuroscience, School of Neuroscience, Department of Basic & Clinical NeuroscienceWebsite

Combining expansion microscopy and CRISPR-tagging to understand amyloid-induced synaptic pathology.

Memory loss in Alzheimer’s disease (AD) is thought to be due synaptic dysfunction and synapse loss, pathogenesis initiated by oligomeric amyloid beta. However, there is currently a paucity in our understanding of what happens to not only the structure of synapses, but also to the proteins that localise to them. Moreover, while we know that synapses change shape and number in response to synaptic stimuli, how this is altered in AD is also unclear.
One of the major challenges of studying synapses is the ability to accurately resolve the morphology of synapses as well as the protein that reside within them. This is because the size of synapses falls below the resolution limits of traditional microscopy. A second challenge is the ability to label synaptic proteins in order to visualise the proteins and to subsequently assess their nanoscopic localisation.
Advances in optical microscopy techniques has led to the development of expansion microscopy (ExM) – an approach that allows nanoscale imaging of biological specimens through the physically magnification of biological specimen. Advances in the field of genome editing, and specifically the development of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 approaches now allows for the tagging of endogenous proteins in living cells.
This PhD project aims to combine both ExM with CRISPR/Cas9-tagging of synaptic proteins to examine synaptic structures and proteins at the nanoscale. The first part of the project will focus on optimising ExM and CRISPR-Cas9-tagging approaches to visualise synaptic proteins in primary rodent neuronal cultures, ex vivo brain tissue as well as human iPSC-derived neurons. Subsequently, these approaches will be used in combination to examine the nanoscale consequence of amyloid-driven synaptotoxicity, and to subsequently examine how activity(synaptic)-dependent remodelling of synapses is altered in AD.

Representative Publications

Rupawala H, Shah K, Davies C, Rose J, Colom-Cadena M, Peng X, Granat L, Aljuhani M, Mizuno K, Troakes C, Gomez Perez-Nievas B, Morgan A, So P-W, Hortobagyi T, Spires-Jones TL, Noble W, Giese KP (2022). Cysteine string protein-alpha accumulates with early pre-synaptic dysfunction in Alzheimer’s disease. Brain Commun 4:fcac192. doi: 10.1093/braincomms/fcac192

Martinez-Serra R, Alonso-Nanclares L, Cho K, Giese KP (2022). Emerging insights into synapse dysregulation in Alzheimer’s disease. Brain Commun 4:fcac083. doi: 10.1093/braincomms/fcac083

Ghosh A, Mizuno K, Tiwari SS, Proitsi P, Gomez Perez-Nievas B, Glennon E, Martinez-Nunez RT, Giese KP (2020). Alzheimer’s disease-related dysregulation of mRNA translation causes key pathological features with ageing. Transl. Psychiatry 10:192. doi: 10.1038/s41398-020-00882-7.

Sheppard PAS, Chandramohan D, Lumsden A, Vellone D, Denley MCS, Srivastava DP, Choleris E. Social memory in female mice is rapidly modulated by 17?-estradiol through ERK and Akt modulation of synapse formation. PNAS (in Press) –

Erli F, Palmos AB, Raval P, Mukherjee J, Sellers KJ, Gatford NJF, Moss S, Brandon N, Penzes P, Srivastava DP. “Estradiol reverses excitatory synapse loss in a cellular model of neuropsychiatric disorders.” Translational Psychiatry 2020 Jan 21;10(1):16. doi: 10.1038/s41398-020-0682-4. PMID: 32066698.

Sellers KJ, Elliott C, Jackson J, Ghosh A, Ribe E, Rojo AI, Jarosz-Griffiths HH, Watson IA, Xia W, Semenov M, Morin P, Hooper NM, Porter R, Preston J, Al-Shawi R, Baillie G, Lovestone S, Cuadrado A, Harte M, Simons P, Srivastava DP, Killick R. “?-amyloid synaptotoxicity is Wnt-PCP-dependent and blocked by fasudil.”, Alzheimer’s and Dementia 2018 Mar;14(3):306-317. doi: 10.1016/j.jalz.2017.09.008. Epub 2017 Oct 19. PMID: 29055813.