Investigation of neuronal activity in a murine model of Alzheimer’s disease using in vivo two-photon calcium imaging

Korzhova, Viktoria (2019): Investigation of neuronal activity in a murine model of Alzheimer’s disease using in vivo two-photon calcium imaging. Dissertation, LMU München: Graduate School of Systemic Neurosciences (GSN)

Abstract

Alzheimer’s disease (AD) is one of the biggest challenges for biomedical research nowadays as with the growth of life span more and more people are affected by this disorder. Etiology of AD is unknown, yet growing evidence identifies alterations in neuronal activity as of the great importance for pathology. Although several significant studies of neuronal activity alteration in AD were done during the last decade, none of them addressed the question of the time course of these changes over the disease progression. Alzheimer’s disease (AD) is characterized by impairments of brain neurons that are responsible for the storage and processing of information. Studies have revealed decrease in the activity of neurons (Silverman et al., 2001; Prvulovic et al., 2005) and it was proposed that generalized hypoactivity and silencing of brain circuits takes place as formulated in the synaptic failure hypothesis (Selkoe, 2002). However, more recent studies also reported opposite effects – hyperexcitability and hyperactivity of neurons in the AD models (Busche et al., 2008; Sanchez et al., 2012; Liebscher et al., 2016). It still remains unclear if these are two sides of the same coin or if these are two stages, that follow each other. Moreover, it is not clear if observed neuronal activity alterations are caused by the dysfunction of individual neurons or if overall circuitry is disturbed because the crucial “activity controllers” (most probably - inhibitory neurons) alter their activity. This project aimed to examine spontaneous neuronal activity in the murine model of AD at the early stages of disease progression using chronic in vivo imaging to address the character and the stability of neuronal activity alterations as well relation of the activity alterations to amyloid plaque proximity. Compared to earlier studies the approach of in vivo awake calcium imaging used in the current study has many benefits for brain research. The main advantage is that brain activity can be measured without artifacts generated by anesthesia, which can exaggerate or mitigate experimental readouts. In this project, I used genetically encoded calcium indicator GCaMP6 that enables prolonged repetitive imaging of the same neurons in an intact environment. Recording of calcium transients in cell bodies of neurons was accompanied by in vivo imaging of Aβ plaques and followed by immunohistochemical staining of GCaMP6-expressing neurons to investigate how activity changes are correlated with proximity to the plaque. All the experiments were done in awake mice to ensure the absence of anesthesia-derived impact on spontaneous neuronal activity. My results support previously published reports of the increased proportion of hyperactive excitatory neurons in the AD mouse model. Importantly, my results also demonstrate that this increased activity is present in the awake state, is stable over a longer period of time (one month) and does not depend on the distance to the closest plaque. These findings support the hypothesis of permanent network alterations driving aberrant activity patterns that appear early in the disease progression, resulting in a chronic excitation/inhibition disbalance. Another important finding of my project is that individual neurons do not stay in the silent state and most of them remain functional demonstrating normal activity at the later time points. This finding requires further research as it has important implication for the development of the AD treatment, as in case many neurons remain functional and their normal neuronal activity can be recovered by addressing the cause of the circuit dysfunction with treatment. To summarize, the study presented in this PhD thesis is the first longitudinal study of neuronal activity changes in an AD mouse model, and while it provides important insight into pathology, it also emphasizes the importance of chronic in vivo studies to investigate neuronal activity and its role in the disease progression.

• Dokumententyp: Dissertationen (Dissertation, LMU München)
• Keywords: Alzheimer’s disease, two-photon calcium imaging, in vivo calcium imaging, Alzheimer’s disease mouse model, amyloid pathology, neuronal activity in Alzheimer’s disease
• Themengebiete: 500 Naturwissenschaften und Mathematik ; 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
• Fakultäten: Graduate School of Systemic Neurosciences (GSN)
• Sprache der Hochschulschrift: Englisch
• Datum der mündlichen Prüfung: 11. September 2019
• 1. Bericht­erstatter:in: Herms, Jochen