Alzheimer’s disease (AD) is the most common form of dementia affecting approximately 35 million people globally. Current treatments only reduce symptoms but not the disease process. To elucidate the molecular and biochemical mechanisms underlying AD, and to identify new therapeutic targets, preclinical research often uses transgenic mouse models with characteristics of AD including abnormalities in two neurological proteins, APP and tau. These abnormalities lead to the development of plaques containing aggregated beta-amyloid and tangles of hyperphosphorylated tau. In vitro mammalian neuronal cultures do not spontaneously develop these features and cannot be maintained for extended periods of time in culture, limiting their usefulness for some preclinical research.
Why we funded it
This PhD Studentship aims to partially replace mice used in the study of AD by fully characterising an organotypic slice culture model derived from a transgenic mouse line.
In 2010, 722 papers were published using transgenic mouse models for AD with an average of 23 mice used for experiments in each paper. This equated to a total of approximately 17,000 transgenic mice required worldwide for AD studies published and registered in Pubmed in 2010. To reduce their use, the slice culture method generates approximately 36 slices from one postnatal mouse brain and allows each slice to be maintained in culture for up to six months. This allows for AD-like features to develop and the investigation of a number of parameters/treatments in tissue obtained from the same animal.
Organotypic slices will be prepared from 3xTg-AD mice. These transgenic mice recapitulate many biochemical, behavioural and physiological features of human AD, including the progressive development of plaques and tangles. Using previously published methodology for making organotypic slice cultures, this project aims to fully characterise the biochemical and physiological disease phenotype that develops in the cultured slices with time. This includes understanding the relationship between changes in tau and amyloid-beta with disease progression. Following characterisation, the slice cultures will be validated for their use in drug discovery. This will be achieved by testing the effects of therapeutic agents previously investigated in 3xTg-AD mice and comparing effects on tissue slices and those found in vivo.
Croft CL and Noble W (2018). Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease. F1000Research 7:592. doi: 10.12688/f1000research.14500.2
Croft CL et al (2017). Membrane association and release of wild-type and pathological tau from organotypic brain slice cultures. Cell Death & Disease 8(3): e2671. doi: 10.1038/cddis.2017.97
Croft CL, Kurbatskaya K, Hanger DP, Noble W (2017). Inhibition of glycogen synthase kinase-3 by BTA-EG4 reduces tau abnormalities in an organotypic brain slice culture model of Alzheimer’s disease. Scientific Reports 7:7434. doi: 10.1038/s41598-017-07906-1
Kurbatskaya K, Phillips EC, Croft CL, Dentoni G, Hughes MM, Wade MA, Al-Sarraj S, Troakes C, O'Neill MJ, Perez-Nievas BG, Hanger DP, Noble W (2016). Upregulation of calpain activity precedes tau phosphorylation and loss of synaptic proteins in Alzheimer's disease brain. Acta Neuropathologica Communications 4:34. doi: 10.1186/s40478-016-0299-2
Atherton J, Kurbatskaya K, Bondulich M, Croft CL, Garwood CJ, Chhabra R, Wray S, Jeromin A, Hanger DP, Noble W (2014). Calpain cleavage and inactivation of the sodium calcium exchanger-3 occur downstream of Aβ in Alzheimer's disease. Aging Cell 13(1):49-59. doi: 10.1111/acel.12148
Phillips EC, Croft CL, Kurbatskaya K, O'Neill MJ, Hutton ML, Hanger DP, Garwood CJ, Noble W (2014). Astrocytes and neuroinflammation in Alzheimer's disease. Biochemical Society Transactions 42(5):1321-5. doi: 10.1042/BST20140155.