Presenter of 1 Presentation
ACETYLCHOLINESTERASE IN IPS-DERIVED CORTICAL NEURONS FROM AD-PATIENT.
Abstract
Aims
Acetylcholinesterase (AChE) is the enzyme that hydrolyses acetylcholine at cholinergic synapses. In Alzheimer’s Disease (AD), despite the decrease in enzymatic activity it has been described maintained levels of AChE protein in cortical areas. Furthermore, non-cholinergic roles have been described for AChE like favouring neurite outgrowth or amyloid beta deposition. In order to reflect changes in AD brain, culture of human induced pluripotent stem cells (iPS)-derived cortical neurons provide a great cellular model. In this context, the aim of this study is to characterise expression of AChE in patient-derived iPS and neurons when cortical neurons are cultivated alone, co-cultured with astrocytes or/and microglia.
Methods
Differentiations are carried out up to 70 days, maintained in neural maintenance media or BrainPhys, which supposedly favours neuronal maturation. Co-cultures with microglia are performed 14 days before deadline. Then, they are collected for imaging, enzymatic activity, protein and transcript levels.
Results
AChE enzymatic activity increases during differentiation from iPS to cortical neurons. In addition, cortical neurons co-cultured with microglia or when their maturation has been favoured, display an increased AChE activity compared to control conditions. Also, most of the cortical neurons display cholinergic phenotype as shown by immunostaining.
Conclusions
Differentiation of iPS to cortical neurons shifts AChE towards the cholinergic phenotype. Furthermore, there seem to be a more cholinergic form of AChE when iPS-derived cortical neurons are co-cultured with microglia. Therefore, iPS-derived cortical neurons provide a useful cellular model to characterise AChE in AD.