| dc.description.abstract | Alzheimer’s disease is a slowly progressing and ultimately fatal neurodegenerative disease affecting more than 46 million people worldwide in 2018. The entorhinal cortex (EC), in particular the layer II of the domains located towards the collateral/rhinal fissure, contains neurons that are among the very first to undergo pathological alterations associated with the disease. Specifically, neurons in this domain develop the initial cortical tau pathology, while layer II is also known to exhibit severe neuronal loss already during the pre-clinical stages of AD. Furthermore, layer II-neurons are also subject to early accumulation of intracellular Aβ. In order to help enable the study of these early neuronal pathologies the work in this thesis was aimed at developing a platform for studying the LEC layer II neuronal population in vitro. Here, we have developed a protocol for the surgical extraction and culturing of adult LEC layer II-neurons from an AD-phenotype mouse model. Fourteen different experiments were performed to optimize the protocol being able to achieve high neuronal viability. My results demonstrate that it is possible to 1) identify LEC layer II-neurons on fresh vibratome sections, 2) selectively dissect these out while maintaining tissue integrity, and 3) dissociate and culture such neurons to the point that they re-form structural connections. Neuronal attachment was obtained up to two weeks after plating of neurons, and increasing complexity of the LEC layer II-neurons in vitro was observed, where the development of structural connections was achieved. In conclusion, culturing of LEC layer II-neurons is possible, though, future work should focus on optimizing the in vitro protocol to increase the viability of neurons and enable the measurement of neuronal activity. | |
