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P1‐133: Functional Acetylcholine receptors are expressed in human embryonic stem cell‐derived neurons
Author(s) -
Nilbratt Mats,
Porras Omar,
Marutle Amelia,
Hovatta Outi,
Nordberg Agneta
Publication year - 2008
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2008.05.720
Subject(s) - cholinergic , cholinergic neuron , biology , neuroscience , basal forebrain , choline acetyltransferase , acetylcholine , neurotrophic factors , muscarinic acetylcholine receptor , microbiology and biotechnology , nicotinic agonist , receptor , endocrinology , biochemistry
Background: Alzheimer s disease is associated with degeneration of basal forebrain cholinergic neurons (BFCNs) which are very important for cognitive processes such as attention, learning and memory. These disturbances in cognitive functions are correlated with the loss of nicotinic acetylcholine receptors (nAChR). We aimed to generate specific neurogenic progenitors from human embryonic stem (hES) cells and to determine in vitro conditions for their differentiation into a cholinergic phenotype. Since BFCNs originate from the ventral telencephalon it is also important to examine the subregional identity of hES cell-derived neuronal progenitors. Methods: The expression of transcription factors, radial glial cell markers, neuronal markers and cholinergic receptors was analyzed by RT-PCR and immunocytochemistry. The functionality of cholinergic receptors was measured by calcium imaging. Results: Here, we demonstrate that hES cells differentiate into Pax6, GFAP and brain lipid-binding protein immunopositive radial glial cells and then further into III-tubulin and MAP2-positive neurons. Moreover, transcription factors including Mash1, Gsh2 and Dlx2 were expressed specifying a telencephalic lineage. The neurons also expressed the cholinergic enzyme choline acetyltransferase and transcripts for 3, 4 and 7 nAChR subunits and for M1, M2 and M3 muscarinic acetylcholine receptor (mAChR) subtypes. In addition, the cholinergic receptors mediated acetylcholine-evoked increase in cytosolic calcium levels, demonstrating their functionality. This response was abolished by the mAChR antagonist atropine. As neurotrophic factors, including brain-derived neurotrophic factor and nerve growth factor have regulatory roles in neural development, we will also examine their effects on the expression and functionality of various nAChR and mAChR subtypes. Conclusions: Our findings demonstrate expression of functional cholinergic receptors on hES cell-derived neurons, which may facilitate the screening of novel cholinergic drugs.