Investigation of purinergic receptors in human BE(2)-M17 neuroblastoma cells and their role in neurotransmitter release

Rayment, Cameron (2024) Investigation of purinergic receptors in human BE(2)-M17 neuroblastoma cells and their role in neurotransmitter release. Masters thesis, University of East Anglia.

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Abstract

Adenosine 5′-triphosphate (ATP) is a neurotransmitter of postganglionic sympathetic nerves and non-adrenergic, non-cholinergic nerves of the enteric nervous system. Norepinephrine (NE) is a key monoamine neurotransmitter in the central nervous systems and peripheral organs of vertebrate organisms. ATP and NE are co-stored in synaptic vesicles in sympathetic nerves and when co-released act post-junctionally to evoke contraction of visceral and vascular smooth muscle. However, little is known about the effect of ATP on pre-junctional NE release. Here we show that undifferentiated human BE(2)-M17 neuroblastoma cells demonstrate a calcium response to nucleotides with UDP evoking a statistically greater potency and maximal response than ATP. We also successfully transient-transfected undifferentiated human BE(2)-M17 neuroblastoma cells with the GRABNE biosensor, which we used to detect changes in extracellular NE concentrations when exposed to the nucleotides. We isolated the significant difference in the maximal response of NE compared to Epinephrine and Dopamine demonstrating that NE had a greater efficacy on the GRABNE biosensor. However, the EC50 between the three catecholamines was not significantly different suggesting that the catecholamines are equipotent to the GRABNE biosensor. Finally, we found no significant difference in the change of fluorescence in GRABNE transfected cells following exposure to ATP and UDP. Thus, suggesting nucleotides do not excite NE release but may inhibit it.

Item Type:	Thesis (Masters)
Faculty \ School:	Faculty of Science > School of Biological Sciences
Depositing User:	Nicola Veasy
Date Deposited:	02 Jul 2024 09:11
Last Modified:	02 Jul 2024 09:11
URI:	https://ueaeprints.uea.ac.uk/id/eprint/95746
DOI:

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